ACA 2013 Abstract Export
Plenary Session: Plenary Session 1: Novel aspect of therapeutics of autoimmune diseases
011 Vitamin D and rheumatic diseases
S.S. Yeap1
1Department
of Medicine, Sime Darby Medical Centre Subang Jaya, Subang Jaya, Malaysia
Vitamin D is a fat-soluble hormone that is responsible for calcium homeostasis. However, it has paracrine and
immune-modulatory effects through the widespread presence of the vitamin D receptor (VDR) in many cells including
macrophages, dendritic cells, B and T lymphocytes and neutrophils. VDR activation in these cells leads to
transcription of gene products that initiate a cascade of anti-proliferative, pro-differentiating and immune-regulatory
processes. Dendritic cells are likely to be the primary immune target of 1,25(OH)D, affecting its role in antigen
processing and presentation. Overall, 1,25(OH)D appears to suppress cell-mediated (Th1 and Th17) processes, and
promote a regulatory or tolerogenic environment that may act to ÔcontainÕ autoimmune pathological mechanisms.
Thus a lack of vitamin D may exacerbate autoimmune pathology.
The largest number of studies with regards to vitamin D and autoimmune diseases has been done in systemic lupus
erythematosus (SLE) and rheumatoid arthritis (RA). There is evidence that low vitamin D levels may increase disease
susceptibility to both SLE and RA. SLE patients with low 25(OH)D levels have lower bone density, and increased
SLEDAI scores. In RA, low 25(OH)D levels have been found to be negatively correlated with disease activity e.g. as
measured with the Disease Activity Score (DAS28). Low 25(OH)D values were also found in those not in remission or
responding poorly to treatment. However, there have been no clinical trials formally looking at vitamin D
supplementation in the prevention or treatment of rheumatological disorders.
012 NK cells sustain immune tolerance at human maternal-fetal interface
Z.G. Tian1, B.Q. Fu1, X.C. Li2, R. Sun1, H.M. Wei1
1University
of Science & Technology of China, School of Life Sciences, Hefei, China
and Women’s Hospital and Harvard Medical School, Transplant Research Center, Boston, USA
Natural killer (NK) cells are innate immune lymphocytes that can control several types of tumours and microbial
infections. Current research highlights the fact that natural killer cells also act as regulatory cells to affect adaptive
immune response. Interestingly, NK cells accumulate in large numbers at the maternal-fetal interface, but their exact
roles in successful pregnancy remain poorly defined. Here, we provide evidence that TH17 cells and local
inflammation can occur at the maternal-fetal interface during natural allogenic pregnancies. We found that decidual
NK cells promote immune tolerance and successful pregnancy by dampening inflammatory TH17 cells via IFN-γ
secreted by CD56brightCD27+ NK subset. We also found that this NK cell-mediated regulatory response is lost in
patients who experience recurrent spontaneous abortions, which results in a prominent TH17 response and extensive
local inflammation. This local inflammatory response further impacts the regulatory function of NK cells, leading to the
eventual loss of maternal-fetal tolerance. Thus, our data identify NK cells as key regulatory cells that sustain immune
tolerance at the maternal-fetal interface by suppressing TH17-mediated local inflammation.
2Brigham
Parallel Session: Parallel Session 1: Epidemiology and clinical presentation of autoimmune diseases
018 Antiphospholipid syndrome: lessons from the genetic study
Genetic factors are hypothesized to play a role in the susceptibility to antiphospholipid syndrome (APS) based on
several family studies in patients with antiphospholipid antibodies (aPL) and/or clinical manifestations of APS. The
genetics of β2-glycoprotein I (β2GPI), one of the most representative target antigens of aPL, has been extensively
studied. 247 Val/Leu polymorphism can affect the conformational change of β2-GPI and the exposure of the epitopes
for aCL. We found that 247 Val was correlated with anti-β2-GPI production in patients with primary APS, and 247 Val
may be important for β2-GPI antigenicity. STAT4 SNP in Japanese patients with SLE and/or APS. T allele
frequencies in SLE and APS were significantly elevated compared with that in healthy controls. When analyzed only
in primary APS patients, T allele frequency was further higher. BANK1, BLK and SNP in 1q25.1 region were
associated with not only SLE but also APS in Japanese population. These results suggest that APS and SLE, in part,
share a common genetic background.
Although the genes involved in APS have not been identified because antigen specificity of aPL and the
pathophysiology of APS are highly heterogeneous and multifactorial, genomewide linkage analysis and large cohort
studies would lead to better understanding of the genes that might be involved in APS.
020 DEVELOPMENT OF PATIENT-BASED RA DISEASE ACTIVITY INDEX
M.H. Leung1
1Medicine,
Queen Elizabeth Hospital, Hong Kong, Hong Kong China
Standard disease activity measures in rheumatoid arthritis such as the Disease Activity Score in 28 joints (DAS28)
are based on evaluators and have high inherent inter-observer variability. Other commonly used simplified versions
are improved mainly in the arithmetic. The other limitation is the dependency for blood tests on inflammatory indices,
the result of which may not be readily available upon clinic visit. Moreover, patients cannot monitor their own disease
activity at home, in a way similar to ambulatory blood pressure and home haemoglucostix monitoring in other chronic
illnesses.
Patient-based disease activity score (PDAS1 [with erythrocyte sedimentation rate ESR] & PDAS2 [without ESR]) in
rheumatoid arthritis (RA) are developed in an attempt to overcome these limitations. In the developmental cohort, the
PDAS with ESR (PDAS1) and without ESR (PDAS2) achieved excellent fit with the DAS28. The results were
validated in another validation cohort. The PDAS and DAS28 had similar sensitivity to change, determined using
effect sizes and standardized response means.
Status and responder criteria of European League Against Rheumatism (EULAR) DAS28 had also been mapped to
PDAS. Remission, low, moderate and high disease activities, good and moderate responses to treatment were
determined. They have comparable agreement and performance to standard criteria based on DAS28 and Clinical
Disease Activity Index (CDAI).
PDAS appears suitable for clinical decision-making, epidemiologic research, and clinical trials.
021 Introduction of Korean nationwide RA cohort and Clinical implication
Sang-Cheol Bae, MD, PhD, MPH
Department of Rheumatology, Hanyang University Hospital for Rheumatic Diseases, Seoul 133-792, Korea
A large national prospective cohort can be crucial in performing clinical research. In South Korea, KORean
Observational study Network Arthritis (KORONA) has been established by Clinical Research Center for Rheumatoid
Arthritis (CRCRA) at Hanyang University funded by the Ministry of Health and Welfare in 2008. KORONA has been
investigated by using prospective protocol including patientsÕ demographics, clinical features, laboratory and
radiologic data, health-related outcomes, treatment side effects, resource utilization, and health behaviors in 23
University Hospitals. CRCRA have implemented annual monitoring, auditing, staff training and provides standard
operation protocol. Currently, 5,376 RA patients were enrolled and approximately 80% of their follow-ups are going
on.
Prospective cohort study can provide the information that cross-sectional study or randomized clinical trials (RCTs)
cannot provide and it also enables to reflect clinical practice. For example, it enables researchers to investigate how
patientsÕ characteristics (i.e. genetic factors) could affect the disease outcomes and to investigate both research
subjects that cannot be conducted by RCTs and excluded patients in RCTs (i.e. patients with old age or comorbidities
who excluded in RCT). In addition, cohort study can reflect real clinical practice and provide long-term patientsoriented outcomes (PROs).
In this talk, I would like to introduce KORONA cohort and present the important results of our studies using this
cohort.
022 IS ATHEROSCLEROSIS AN AUTOIMMUNE DISEASE?
H. Tse1, X. Zhao1, R. Thummel2, M. Shaw1
1Immunology
and Microbiology, Wayne State University, Detroit, USA
and Cell Biology, Wayne State University, Detroit, USA
Background and Aims: Atherosclerosis is a chronic inflammatory disease of the arterial wall with immune
implications. Both innate and adoptive immunity are involved. Initially, autoantibodies against oxidized low density
cholesterol (oxLDL) are found in the sera of atherosclerotic mice and human. T cells of all subclasses are also
identified at the lesion sites. In addition, studies with gene-deficient mice further demonstrate the role of T cells in the
development of disease. Despite these studies, however, there are no data demonstrating the cause-and-effect
relationship of T cells and atherosclerosis. Specifically, the nature of the atherogenic T cells remains elusive. To
identify atherogenic T cells, we reason that T cells are antigen specific and identification of the atherogenic epitopes
of an autoantigen should lead us to identification of the atherogenic T cells themselves.
Methods: Applying the mouse H-2b-binding motif published by Liu et al., we scanned over the entire 4500 amino acid
sequence of the murine autoantigen, apoB100, and identified 18 sequences that fit the motif characteristics. Six such
sequences were selected for synthesis and for functional studies.
Results: One of the peptides, peptide6, significantly elevated the aortic plaque burden of Western Diet-fed ApoE-/mice immunized with the peptide emulsified in CFA. Lymph node cells from P6-immunized mice and P6-specific T
cell clones also successfully adoptively transferred enhanced atherosclerosis to na•ve recipients fed a WD diet. P6 is
the first ever identified atherogenic T cell epitope of apoB100.
Conclusions: These studies lend strong support to the concept that atherosclerosis is an autoimmune disease.
2Anatomy
023 DETECTION OF OXIDIZED LIPOPROTEINS ACCUMULATED IN ATHEROSCLEROSIS LESIONS BY
IMAGING MASS MICROSCOPY (IMS)
L. Shen1, W. Arum Tri2, K. Kobayashi1, T. Sasaki1, Y. Matsunami2, F. Takenaka1, E.I.J.I. Ando3, T. Yamamoto4, K.
Ogata3, E.I.J.I. Matsuura1
1Collaborative
Research Center (OMIC), Okayama University Graduate School of Medicine Dentistry and
Pharmaceutical Sciences, Okayama, Japan
2Department of Cell Chemistry, Okayama University Graduate School of Medicine Dentistry and Pharmaceutical
Sciences, Okayama, Japan
3Life Science Business Department, Shimadzu Corporation, kyoto, Japan
4Applications Development Center, Shimadzu Corporation, kyoto, Japan
Background: β2-Glycoprotein I (β2GPI) is a major antigen for pro-thrombotic autoantibodies characteristically
present in patients with antiphospholipid syndrome. β2GPI particularly binds to oxLDL via 7-ketocholesteryl-9carboxynonanoate (oxLig-1) to form pro-atherogenic oxLDL/β2GPI complexes. Anti-β2GPI autoantibodies enhance
the uptake of oxLDL/β2GPI by macrophages via Fcγ receptors. We studied the in vitro intracellular accumulation of
lipids by cultured macrophages (J774A.1), and in aortic lesions containing atherosclerotic plaques by MALDI-TOFmass spectrometry (MALDI-TOF-MS) and the novel imaging mass microscope (iMS). Methods: Purified LDL from
LDL-/- mice fed with a high fat diet was oxidized by CuSO4 treatment, and the resulting oxLDL incubated with J774A.1
cells to promote phagocytosis. A profile of oxidized lipids was obtained by MALDI-TOF-MS, and the distribution of
lipids within atherosclerosis plaques was imaged by iMS. Results and Conclusion: A positive ionization mass
spectrum (m/z 369; from cholesterol) was detected in all specimens (aortic tissue and plasma). In contrast, m/z 383
corresponding to the mass size of 7-ketocholesterol appeared in oxidized lipoprotein fractions and in oxLDL-loaded
J774A.1 cells. Finally, 2-dimentional iMS visualized aortic atherosclerotic plaques in the Watanabe heritable
hyperlipidemic rabbits and in LDL-/- mice. Thus, MALDI-TOF-MS and iMS techniques may create new era in
methodology for pathophysiological studies of lipid metabolism.
Parallel Session: Parallel Session 2:Cutting edge in therapy of autoimmune disease
027 Is the proteasome a valuable target for treating autoimmune diseases?
J. Kalden1, R. Voll2
1Division
for Molecular Immunology, University of Erlangen-Nuremberg, Erlangen, Germany
for Rheumatology and Clinical Immunology, University of Freiburg, Freiburg, Germany
Inhibition of the immuno proteasome has been recently demonstrated tohave a potential therapeutic effect especially
in patients sufferingfrom SLE. Studies in experimental animal models for SLE proved to showthat the inhibition of the
proteasome not only prevented thedevelopment of the disease, but also in animals with manifest SLEsyndromes
showed a significant improvement in antibody titors andkidney function. Studies on the function of the inhibition of
theproteasome revealed that specifically long-lived plasma cells will beaffected, and as more recently shown, that the
interferon alphaproduction by plasmacytoid dendritic cells will be significantlyinhibited (H. Travis Ichikawa et al.
Arthritis Rheum 2012, 64:493-503).Severe side effects observed in the experimental animal models wereacceptable.
There was no major influence by the inhibition ofbortezomib on T cell function.
More recently, an open, multicenter trial in SLE patients has beeninitiated in Germany. First data indicate that
Bortezomib is ofclinical efficacy in patients with a severe form of disease includingkidney involvement. However,
more data are necessary to show thebeneficial effect of a inhibition of the proteasome on a long run. Atthe moment it
looks like that the inhibition of the proteasome might bea valuable option for the treatment start and for the treatment
offlaires of SLE patients.
Since bortezomib as shown by the treatment of myeloma patients cancause polyneuropathies new molecules for
inhibiting the proteasome havebeen developed and are presently under clinical investigation.
2Department
028 ADVANCEMENTS IN THE TREATMENTS OF MULTIPLE SCLEROSIS
K.H. Chan1
1Medicine,
The University of Hong Kong, Hong Kong, Hong Kong China
Abstract
Multiple sclerosis (MS) causes neurological disability. Experimental autoimmune encephalomyelitis (EAE) is an
animal model of MS. Human bone-marrow mesenchymal stem cells (hMSCs) possess anti-inflammatory and
immunosuppressive effects. We studied whether hMSCs affect CD1d highCD5+ regulatory B cells activity in EAE. EAE
is induced in C57BL/6N mice by immunization with MOG35-55 peptide. hMSCs are injected intravenously into EAE
mice at day 3 and day 12 after first immunization. Mice were sacrificed at day 26. Immunohistochemistry of spinal
cord, serum cytokines levels, production of cytokines by cultured splenic cells, and flow cytometry for splenic Th17
and CD1dhighCD5+ regulatory B cells were studied. We found that EAE mice with hMSCs treatment as day 3 and day
12 had reduced EAE scores from day 14 to 26 compared to EAE mice without hMSCs treatment, and reduced
infiltration of inflammatory cells and demyelination in spinal cord. EAE mice with hMSCs treatment at day 3 and day
12 had 1) lower serum levels of IL-6, TNF-α (p<0.0005) and IL-17 (p<0.005 for day 3, p<0.0005 for day 12), 2)
reduced splenic cell production and secretion of IL-6, TNF-α (p<0.05) and IL-17 (p<0.05), and increased splenic
production of IL-10, 3) reduced splenic Th17 cells (p<0.05 for day 3, p<0.005 for day 12) and 4) increased
CD1dhighCD5+ regulatory B cells (p<0.005) than EAE mice without hMSCs treatment. hMSCs treatment at day 3 and
12 suppresses EAE severity. The underlying mechanisms involve downregulation of Th17 cells and upregulation of
CD1dhighCD5+ regulatory B cells activity.
029 Can suppression of inflammation prevent progression of premature atherosclerosis in rheumatic
diseases?
L.S. Tam1
1Department
of Medicine & Therapeutics, The Chinese University of Hong Kong, Hong Kong, China
Tumor necrosis factor-α (TNF-α) antagonists are effective in suppressing inflammation in patients with rheumatoid
arthritis (RA), ankylosing spondylitis (AS) and psoriatic arthritis (PsA). However, it is debatable if they have favorable
effects on the cardiovascular (CV) system. The use of surrogate end points instead of actual CV events may provide
more evidence on whether rheumatic disease patients may benefit from potent anti-inflammatory treatment such as
anti-TNF-α therapy on the prevention of premature atherosclerosis. Surrogate markers including carotid intima media
thickness (cIMT) has been widely used to assess subclinical atherosclerosis; while pulse wave velocity (PWV) and
aortic augmentation index (AIx) are the gold standards to assess arterial stiffness. Published data suggest that TNF-α
blockers probably are effective in preventing or even reversing the progression of IMT in patients with RA, AS and
PsA who are responding to treatment. With regards to arterial stiffness, PWV was either significantly reduced or
remained unchanged in most studies following TNF-α antagonist treatment; nonetheless, most studies in RA reported
significant improvement of PWV. In contrast, AIx remained unchanged in the majority of the studies. The balance of
evidence suggests that TNF-α antagonists may have a beneficial effect on preventing the progression of subclinical
atherosclerosis and arterial stiffness. Consequently, this may have a beneficial effect on CV risk. However, longer
term cohort studies are needed to confirm these promising results.
031 HELLP SYNDROME: COMPLICATION OR A NEW AUTOIMMUNE ENTITY?
C. De Carolis1, C. Perricone2, R. Perricone3, F. Crescenzi1
1Obstetrics
and Gynecology, S. Giovanni Addolorata Hospital Rome Italy, Rome, Italy
di Medicina Interna e Specialità Mediche, Reumatologia Sapienza Università di Roma, Rome, Italy
3Dipartimento di Medicina Interna, Rheumatology/Allergology and Clinical Immunology Tor Vergata University of
Rome Rome Italy, Rome, Italy
Recurrent spontaneous abortion (RSA) is defined as two or more consecutive spontaneous abortions, which may
occur at any stage of pregnancy. Among various causes of RSA, autoimmune abnormalities can play a relevant role the most significantly implicated being antiphospholipid antibodies (aPL). However, in several cases, no evident
explanation is found and recurred abortion is reported to be unexplained. A deeper insight into apparently
unexplained RSA shows increasing evidences supporting also an alloimmunologic mechanism, a condition in which
natural killer (NK) cells may play a relevant role when in too high concentrations. Given the poor understanding of the
precise mechanisms underlying RSA and unexplained RSA, therapy so far has been empirical and not evidence
based. Treatment modalities for RSA include aspirin, heparin, progesterone, prednisone and intravenous
immunoglobulins (IVIGs). Different immunomodulatory effects have been reported for IVIG and, among others,
antigen neutralization, Fc-receptor blockade and anti-idiotypic interactions, changes in distribution and function of Tcell subsets, antibodies to autoantigens and mild activation of complement. Furthermore, it is known that IVIGs affect
cytokine production in T lymphocytes and monocytes/ macrophages, all mechanisms of potential benefit in RSA. Our
study compared intravenous immunoglobulins (IVIG) and prednisone in treating pregnant women with a history of
recurrent fetal loss in terms of live-birth rate and maternal and perinatal morbidity. The results of this study will be
discussed.
2Dipartimento
Plenary Session: Plenary Session 2: The environment and autoimmunity
036 Environmental factors in autoimmunity
M. Gershwin1
1Division
of Rheumatology Allergy and Clinical Immunology, University of California at Davis, Davis, USA
PBC is distinguished by immune mediated destruction of the intra hepatic bile ducts and the presence of high titer
antimitochondrial autoantibodies directed against a highly specific epitope within the lipoic acid binding domain of the
pyruvate dehydrogenase E2 subunit. We postulate that chemical xenobiotics modification of the lipoyl domain of
PDC-E2 is sufficient to break self-tolerance, with subsequent production of AMA in patients with PBC. Indeed, using
quantitative structure activity relationship (QSAR) analysis on a peptide-xenobiotic conjugate microarray platform, we
have demonstrated that when the lipoyl domain of PDC-E2 was modified with specific synthetic small molecule lipoyl
mimics, the ensuing structures displayed highly specific reactivity to PBC sera, at levels often higher than the native
PDC-E2 molecule. Hereby, we discuss our recent QSAR analysis data on specific AMA reactivity against a focused
panel of lipoic acid mimic in which the lipoyl di-sulfide bond are modified. Furthermore, data on the immunological
characterization of antigen and Ig isotype specificities against one such lipoic acid mimic; 6,8-bis(acetylthio)octanoic
acid (SAc), when compared with rPDC-E2, strongly support a xenobiotic etiology in PBC. This observation is of
particular significance in that approximately one third of patients who have taken excessive acetaminophen (APAP)
developed AMA with same specificity as patients with PBC, suggesting that the lipoic domain are a target of APAP
electrophilic metabolites such as NAPQI. We submit that in genetically susceptible hosts, electrophilic modification of
lipoic acid in PDC-E2 by acetaminophen or similar drugs can facilitate loss of tolerance and lead to PBC.
Plenary Session: Plenary Session 2: The environment and autoimmunity
038 THE INTERACTOME LIES AT THE HEART OF THE MOSAIC OF AUTOIMMUNITY
T. Marshall1
1Health
Sciences, Murdoch University, Perth, Australia
The Mosaic of Autoimmunity - that seemingly countless diversity in symptoms and disease presentation between
individual autoimmune patients - has long seemed intractable. However, the past few years have seen an explosion
in Molecular Biology's understanding of the complexity of the human body. The Human Interactome is now the name
given to the map of all the interactions between metabolites of the human body. It contains all the proteins (at least
100,000), all the enzymes, peptides and steroids. We can now add to that the thousands of RNAi pathways which
allow RNA to interfere with both genomic and non-genomic functions. But one question still remains: 'Why is there a
Mosaic - Why is there such variability in disease presentation?' The new science of Metagenomics explains that
variability between individuals. We now know the human body is a superorganism * comprising not just the 25,000
genes of Homo Sapiens, but also a million or more genes from the microbes which accumulate in our tissues as we
go through life. Our mother's breast milk contains at least 400 distinct species of bacteria to seed each individual's
microbiome. Throughout life we continue to accumulate microbes from our family, our environment, from our food,
and even from the air we breathe. Some of these microbes have the ability to persist inside the cells of our tissues,
and some can even survive inside the cells of our immune system itself. While common pathogens such as
Mycobacteria and EBV have long been known to be obligate intracellular persisters, the DNA techniques developed
for Metagenomics have shown us that the Human Microbiome consists of thousands, perhaps hundreds of
thousands, of persistant microbial species. Not all of them are 'friendly.' Yet all of them can reduce the ability of our
cells to operate correctly - they affect our Interactome - and ultimately produce that mosaic of dysfunction we know as
'autoimmunity.'
039 INFECTION AND IMMUNITY-MEDIATED ATHEROSCLEROSIS
E. Matsuura1, L. Shen1, L. Lopez2
1Collaborative
Research Center and Department of Cell Chemistry, Okayama University Graduate School of
Medicine Dentistry and Pharmaceutical Sceiences, Okayama, Japan
2R&D, Corgenix, Broomfield, USA
Atherosclerosis is a multifactorial chronic inflammatory disease, advanced by infectious/innate or auto-immunity
accelerating mechanisms. Several epidemiological studies have revealed that host immune responses against
persistent infectious pathogens, such as C. pneumonia, P. gingivalis, H. pylori, and cytomegalovirus, may promote
the development of atherosclerosis. Notably, one mechanism is that immune responses against pathogen-derived
heat-shock proteins that subsequently cross-react with host-derived ones contribute to atherogenesis. Another
autoimmune-associated atherogenic mechanism involves autoantibodies against β2-glycoprotein I (β2GPI). β2GPI
displays pleiotropic roles by influencing fibrinolysis, angiogenesis, and more recently, atherogenesis. In fact,
oxLDL/β2GPI complexes formed in arterial lesions can generate an adaptive immune response (anti-oxLDL and antiβ2GPI antibodies) that up-regulate the macrophage expression of FcγRI and CD36 resulting in an increased uptake
and intracellular lipid accumulation. It was also confirmed that unlike free β2GPI, oxLDL/β2GPI complexes were
quickly incorporated into macrophage lysosomes where pro-inflammatory inflammasome/IL-1 responses are
generated, suggesting a role of β2GPI in innate immunity and chronic inflammation. Similar to adaptive immunity,
dysregulation of the innate immune response may cause tissue injury, chronic inflammation and disease.
Inflammasomes are multi-molecular cytoplasmic complexes of NLRPs that regulate pro-inflammatory caspases and
IL-1 in response to endogenous danger signals. Cholesterol crystals and oxidation specific epitopes (oxLDL) can
activate endogenous NLRP3 inflammasomes to generate pro-inflammatory IL-1β. Thus, an inflammasome-induced
IL-1β overproduction may represent an early atherogenic mechanism that initiates atherosclerosis and, OxLDL and
oxLDL/β2GPI complexes may not only participate in early atherogenesis via the innate inflammasome/IL-1 pathway,
but also perpetuate atherogenesis via an adaptive autoimmune response.
048 GENETICS OF LUPUS: WHAT SHOULD WE KNOW?
C. Perricone1, C. Ciccacci2, F. Ceccarelli3, D. Di Fusco2, E. Cipriano3, G. Novelli2, G. Valesini3, P. Borgiani2, F. Conti3
1Dipartimento
di Medicina Interna e Specialità Mediche, Sapienza Università di Roma, Roma, Italy
Department of Biopathology and Diagnostic Imaging, University of Rome Tor Vergata, Rome, Italy
3Dipartimento di Medicina Interna e Specialità Mediche, Sapienza Università di Roma, Rome, Italy
Systemic Lupus Erythematosus (SLE) is a chronic condition initiated by the loss of immunological tolerance to selfantigens. The pathogenic hypothesis comprises a complex interaction between genetic, environmental and hormonal
factors that interact in an individual over time generating a dysregulation of the immune system leading to disease
development. However, only few genes have been implicated in the contribution to SLE susceptibility. From one side,
recent evidences suggest that a susceptible individual develops autoantibodies over a long time lapse due to
triggering factors. Nonetheless, such autoantibody production is genetically determined and finally, the presence of
autoantibodies can possibly determine the clinical presentation of SLE. Thus, the genetic predisposition to the
development of autoantibodies and toward the disease process may overlap. In this light, the TRAF3IP2 gene,
associated with susceptibility to psoriatic arthritis and psoriasis, encoding for Act1, has been found to be associated
with disease susceptibility, with the occurrence of anti-SSB/La antibodies and with pericarditis. Since this molecule is
a negative regulator of adaptive immunity and a positive signaling adaptor in IL-17 mediated immune responses, it is
clear the contribution of both the innate and adaptive arms of the immune system in establishing SLE risk. Unraveling
these mechanisms will allow to diagnose, to treat and even to prevent the development of SLE.
2Genetics
049 Measurement of flares in SLE
J. Thumboo1
1Department
of Rheumatology and Immunology, Singapore General Hospital/ Dulke-NUS Graduate Medical School,
Singapore, Singapore
A flare of SLE disease activity can be conceptualized as the result of events at a genetic level involving several
genes, which act through various mediators at a cellular level to produce the clinical phenotypes of a SLE flare.
Flares of SLE disease activity are common, ranging from 0.65 to 3.0 flares per patient year in reported series, and
occur in approximately 80% of patients during the course of their disease. The morbidity and mortality associated with
flares can be substantial, and is related to organ damage resulting from active SLE per se and the adverse effects of
corticosteroids and immunosuppressive drugs. For example, doubling of serum creatinine was 7 times more likely in
patients with a renal flare, and 27 times more likely in severe renal flares manifesting as nephritic syndrome. Flares
also result in significant mortality, accounting for 26% of deaths within 5 years of diagnosis and 10% of deaths more
than 5 years after diagnosis in one series. Prediction of flares is thus an important goal in the management of SLE.
However existing clinical and laboratory parameters do not predict flares with sufficient accuracy for clinical use, and
two randomised controlled trials of corticosteroid use in patients with rising double stranded DNA levels or falling
complement levels / rising complement split product levels yielded disappointing results. This highlights the needs for
better markers of flares in SLE.
050 Biomarkers of SLE and Nephritis
Y. Avihingsanon1
1Center
of Excellence in Immunology and Immune-mediated Diseases, Faculty of Medicine Chulalongkorn University,
Bangkok, Thailand
Kidney diseases in SLE are not always immune-complex glomerulonephritis or Òlupus nephritisÓ. Rising of serum
creatinine, microhematuria and proteinuria are non-specific manifestations. Rising of serum creatinine could occur
from drug-induced nephrotoxicity. Microhematuria could be found in kidney calculi or infections. Proteinuria could be
caused by non-SLE glomerulopathy. Therefore, physicians have to make a differential diagnosis and provide different
treatment.
Recent advances in molecular diagnostic technologies have provided new tools in diagnosis and prognosis of lupus
nephritis. Study of both innate and adaptive immune responses provides novel biomarkers of lupus disease activity.
Blood leucocytes from patients with active lupus nephritis were useful for diagnosis and prognostication. Myeloidrelated proteins are interesting markers of innate immune response that associated with active lupus nephritis. Serum
levels of B-cell signatures, like BAFF or APRIL, had been good markers and, perhaps, been targeted for therapy for
SLE and nephritis.
The study of molecular signals from urinary cellular components is a logical approach in diagnosing an early relapse
of nephritis and monitoring response to therapy. Urinary cytokines like interferon-inducible protein-10 (IP-10) and
growth factors like vascular endothelial growth factor (VEGF) in LN are associated with the severity of renal
pathology. Furthermore, the cytokine mRNAs of urine cells has been shown to be associated with response to
therapy. It is likely that early treatment based on urinary molecular signals may alter the course of the disease. This
would support the practicing of individualized medicine in the near future.
051 Biomarkers for early recognition of lupus-related organ injury
A. Mak1
1Medicine,
National University of Singapore, Singapore, Singapore
Along with renal damage, neuropsychiatric and cardiovascular damage has been impacting negatively on the survival
of patients with systemic lupus erythematosus (SLE) for the past few decades. Discrepantly, the substantial advance
in the knowledge of classification, diagnosis, therapeutic paradigm and prognosis of lupus glomerulonephritis has not
been equally observed in neuropsychiatric SLE (NPSLE) and cardiovascular involvement of lupus. The limited
accessibility of neural and vascular tissues for histopathological studies is probably one of the major obstacles to
advance the understanding of the pathogenetic mechanism of NPSLE and cardiovascular disease in SLE.
Fortunately, the recent advent of functional magnetic resonance brain imaging and biophysical endothelial function
assessment has brought researchers non-invasive and sensitive approaches to unravel the functional brain and
vascular pathology of lupus patients respectively, without risking the latter for unnecessary invasive procedures. With
proper standardization and validation of these novel and promising tools amongst research centres, such biophysical
platforms which detect organ injury with a pathogenesis-driven approach will likely add to the armamentarium of
diagnostic and prognostic biomarkers for early lupus-related organ damage, and facilitate the development of
potential therapeutic targets for lupus. Without these, it would be rather difficult to further improve the well-being and
survival of patients with SLE.
052 PATHOGENESIS OF NEUROPSYCHIATRIC LUPUS
T.M. Mok1
1Department
of Medicine, University of Hong Kong, Hong Kong
Systemic lupus erythematosus (SLE) is an autoimmune disease that affects predominantly young women leading to
significant morbidity and mortality. Neurological involvement in SLE may affect the central and peripheral nervous
systems and may present with a wide variety of manifestations from headache to seizure. The underlying
pathogenesis is not as well established as other manifestations of SLE and is likely heterogeneous. Diffuse
neurological involvement often carries poor prognosis. The American College of Rheumatology has set up a task
force in 2000 and proposed 19 classifications of neurological manifestations in SLE in order to standardize the
nomenclature that may facilitate research into the underlying aetiology and treatment of this condition.
053 Infections in SLE
S. Navarra1
1Dept.
of Rheumatology, University of Santo Tomas, Manila, Philippines
Infections are an important cause of morbidity and mortality in systemic lupus erythematosus (SLE). There are
several speculations on the contributory role of microbes in the induction and reactivation of SLE largely invoking the
mechanism of molecular mimicry. Susceptibility to infections in lupus may be explained by several intrinsic and
acquired defects in the immune system - related to the disease itself, or to the immunosuppressive therapies. A broad
spectrum of infections have been reported in SLE, including bacterial, mycobacterial, viral, fungal and parasitic
infections with the respiratory and urinary tract as the most commonly involved sites. Opportunistic infections are
likewise emerging as important causes of mortality. Endemicity plays an important role in augmenting susceptibility to
certain infections like Salmonella and Mycobacterium tuberculosis infections particularly in developing countries.
A major obstacle in the recognition of infection among lupus patients stems from the similarities in their clinical
manifestations with those of SLE, particularly its active state, the atypical presentation of some infections including
the relative absence of inflammatory signs, blunted immune response, and issues regarding infection prophylaxis in
highly endemic areas. Thus, it is important to exert utmost vigilance in the early diagnosis and treatment of these
infections, including prompt evaluation of all fevers and judicious use of immunosuppressive therapy particularly
steroids.
Parallel Session: Parallel Session 4: Pathological avenues in autoimmunity
056 Immunodeficiency and autoimmunity
M. Gershwin1, C. Chang2
1Division
of Rheumatology Allergy and Clinical Immunology, University of California at Davis, Davis, USA
Dupont Hospital for Children, Thomas Jefferson University, Davis, USA
IgA is the most abundant immunoglobulin in the human body, and performs a very specialized role which involves
mucosal immunity, development of tolerance and protection against infection. IgA is the key immunoglobulin in the
respiratory and gastrointestinal tracts, which provide the most intimate interface between the environment and self.
Normal levels of IgA are based on early studies consisting of only small numbers of patients. The international
consensus definition of IgA deficiency is a level of 0.07 g/ml after the age of four years in the absence of IgG and IgM
deficiency. The epidemiology of IgA deficiency reveals interesting variances between geographical regions * the
incidence in Caucasians being much higher than that in Asians. IgA deficiency has also been found to co-exist with
autoimmune diseases, allergies and malignancies. The association with autoimmunity is particularly interesting
because it suggests a common genetic linkage that could potentially also explain the diversity in geoepidemiology.
Both MHC and non-MHC associations have been described and the 8.1 haplotype has been significantly associated
with autoimmunity in IgA deficiency patients over controls. Non-MHC genetic associations include IFIH1 and
CLEC16A. The mutations leading to IgA deficiency have not been defined, but in some cases of IgA deficiency it has
been suggested that the pathogenesis involves a failure in switched memory B cells that can lead to this cohort
experiencing an increased incidence of recurrent bacterial infections or autoimmune diseases. Attempts to investigate
the role of cytokines that can induce IgA synthesis in cells of patients with IgA deficiency, such as IL21 or the
combination of CD40L/anti-CD40, IL-4 and IL10, are underway.
2Nemours/A.I.
058 PATHOLOGICAL IMPLICATION OF AUTOANTIBODIES AGAINST BETA2-GLYCOPROTEIN I AND A
NOVEL “IMMUNO-THERANOSTICS” IN ATHEROSCLEROSIS
E. Matsuura1, L. Lopez2
1Collaborative
Research Center and Department of Cell Chemistry, Okayama University Graduate School of
Medicine Dentistry and Pharmaceutical Sceiences, Okayama, Japan
2R&D, Corgenix, Broomfield, USA
It has been recently established that autoantibodies directed to β2-glycoprotein I (β2GPI) from patients with
antiphospholipid syndrome increase the risk for atherosclerotic disease. The premature development of
atherothrombosis may be explained by IgG immune complexes of β2GPI/oxidized LDL (oxLDL) that are avidly taken
up by macrophages via Fcγ receptors, leading to the formation of laden-foam cells. The possible pathological
mechanism has supported our recent studies: 1) Atherosclerotic plaques in arterial tissue section taken from LDL -/mice and WHHL rabbits were visualized by a novel imaging mass microscope (iMS) detecting lipid ligands for β2GPI.
2) Sudan IV positive-atherosclerotic lesions can be visualized by an in vivo PET/CT imaging using an anti-β2GPI
scFv (lacking the Fc region; 3H3-Fv). Another set of observations using PET suggests that ectopic angiogenesis can
be also detected using a β2GPI variant probe. 3) From the clinical point of view, oxLDL/β2GPI complexes and
antibodies to the complexes frequently appear in patients with autoimmune, diabetes and/or cardiovascular disease
(CVD), associated with vascular complications implicating β2GPI/oxLDL complexes as relevant atherogenic antigens.
Current evidence points toward the atherosclerotic lesions as the primary site of oxLDL/β2GPI complex formation
with subsequent targeted by a humoral immune surveillance. Finally, we report progress in the development of a
novel medical system, namely, ÒTheranosticsÓ (a portmanteau of therapeutics and diagnostics) using a radiolabeled
3H3-scFv (humanized) bearing drug-containing microparticles. Thus, appropriate immunologic PET probes seem not
only to be feasible but of great impact in diagnosis, prevention and management of autoimmunity and CVD.
059 Neonatal autoimmunity and autoinflammatory syndromes
Neonatal lupus is the prototypical neonatal autoimmune disease. It is associated with the presence of antibodies
directed against the nucleoproteins Ro and La in the mother. However, the presence of these antibodies alone is not
sufficient to cause the clinical manifestations of the disease, which include congenital heart block, rash, hepatitis, and
hematologic abnormalities. Besides neonatal lupus, other neonatal autoimmune disease have also been reported,
including neonatal anti-phospholipid syndrome, neonatal myasthenia gravis, neonatal polymyositis and others. The
pathogenic mechanisms of neonatal autoimmunity is unknown, and may involve several different pathways involving
apoptosis of cardiac cells, maternal microchimerism, the presence of cross-reactive antibodies, a dysregulation of T
cells and an abnormality of inhibitory receptors. Unlike neonatal autoimmunity, the pathogenesis of neonatal
autoinflammatory diseases is much better characterized. There are three different forms of the disease, which is
collective known as cryopyrin-associated periodic syndromes (CAPS). The three forms are familial cold
autoinflammatory syndrome (FACS), Muckle-Wells syndrome and neonatal onset multisystem inflammatory disease
(NOMID). All involve a gain-of-function mutation in the cryopyrin gene, CIAS1 or NALP3, which leads to a
dysfunctional inflammasome and a caspase-1 dependent activation of pro-inflammatory cytokines, IL-1b and IL18.
The three diseases were discovered separately, but were later found to have the same genetic mutation. The clinical
presentation of the three diseases is variable, with FACS being the mildest and NOMID the most severe. Recently,
the anti-IL1b drugs, anakinra, canakinumab and rilonacept, have been developed and have had a significant impact
on the treatment of these diseases.
061 DETECTION EIGHT CYTOKINES IN SLE PATIENTS USING MULTIPLEX IMMUNOASSAY
Y. Zhizhong1, H. Weizhen1, Y. Zhihua1, Z. Huiqiong1, C. Xinpeng1
1Rheumatology,
Rheumatology Institute Of Guangdong Medical College, Shenzhen, China
Objectives: Measure the serum levels of eight cytokines in SLE patients and healthy controls using multiplex
immunoassay. Explore the cytokinesÕfunction in SLE and the relationship between cytokines and clinic characteristic
of SLE.
Methods: Serum was separated from peripheral blood of 80 SLE patients and 40 healthy individuals. Multiplex
immunoassay was performed followed the manual of MILLIPLEX MAP human cytokine detecting kit. Measurements
and data analysis of all assays were performed with the Luminex system in combination with the Bio-Plex Manager
software. 0.01 pg/mL cytokine level in serum would be detected. Levels of cytokines were expressed as median(25th,
75th).
Mann-Whitney test was used for all 2 sample comparisons(SPSS, version 13.0). P<0.05 was considered significant.
Results: Without any stimulation, IP-10 was expressed the highest in the eight cytokines, the second was IL-8, the
third was TNF-α. The level of IL-17 was too low to be detected. Only 8 of SLE and 1 of healthy individuals was
measured minim level. The level of IP-10, IL-8 and TNF-α were higher in the SLE patients than that in the healthy
control individuals(P <0.01)(Table 1). The level of IP-10 was positively related with SLEDAI score and the level of
TNF-α was negatively related with C3 level.
Conclusions: The serum level of IP-10, IL-8 and TNF-α were higher in the SLE patients than that in the healthy
control individuals. The three cytokine may play important roles in the pathogenesis of SLE. Multiplex immunoassay
is a reliable, fast, and reproducible technique to measure multiple cytokines with small volumes.
Free Communication: Free Communications 1- Pearls in SLE, APS and RA research
064 AUTOIMMUNITY ACCELERATES ATHEROGENESIS: OXIDIZED-LDL/B2-GLYCOPROTEIN I COMPLEXES
AND ANTI-B2-GLYCOPROTEIN I ANTIBODIES ENHANCE THE RISK OF CARDIOVASCULAR EVENTS IN
ACUTE CORONARY SYNDROME
L.R. Lopez1, T.P. Greco2, B. Hurley3, M. Boisen3, E. Matsuura4
1Clinical
Affairs, Corgenix Inc., Broomfield, USA
Saint Mary's Hospital and Yale University, Waterbury, USA
3Research and Development, Corgenix Inc., Broomfield, USA
4OMIC and Cell Chemistry, Okayama University Graduate School of Medicine Dentistry and Pharmaceutical
Sciences, Okayama, Japan
Background and Aims: OxLDL/β2GPI complexes have been implicated as relevant pro-atherogenic autoantigens and
associated with poor cardiovascular adverse events (AE) by triggering autoimmune pro-coagulant and proatherogenic mechanisms. Because anti-β2GPI antibodies accelerate the in vitro macrophage uptake and intracellular accumulation of oxLDL, the concurrent significance of oxLDL/β2GPI and antiphospholipid antibodies (aPL) on
AE was evaluated in 339 acute coronary (ACS) patients. Methods: Consecutive ACS patients (both genders between
35-75 years) admitted to an acute care center with chest pain and suspected coronary syndromes were studied. The
severity of coronary disease was assessed by angiography. The mean follow-up for AE was 2 years. Serum
oxLDL/β2GPI complexes and aPL were measured by ELISA. Results: AE increased significantly in aPL+ patients
with severe disease than aPL* patients (30.6% vs. 9.0%, p<0.005). Relative risk (RR) for AE of all aPL+ versus aPL*
was 3.66 (95% CI 1.88-7.14, p<0.001). APL+ with oxLDL/β2GPI in 2 upper quartiles had more AE than aPL* in same
quartiles (26.1% vs. 6.0%, p<0.05; RR 3.2, 95% CI 0.95-10.86, p<0.05). RR in aPL+ in 2 upper quartiles was 6.72
(95% CI 2.4-19.0, p=0.01) than aPL* in 2 lower quartiles. RR of aPL+ in the upper quartile was 13.68 (95% CI 1.8102.4, p<0.001) than aPL* in lowest quartile. Conclusions: These results support the concept that the co-presence of
oxLDL/β2GPI complexes and autoimmune aPL antibodies (anti-β2GPI and anti-oxLDL/β2GPI) significantly enhances
the risk of cardiovascular AE, likely mediated by enhancing the intracellular lipid accumulation by macrophage and
atherosclerotic plaque development via scavenger and Fcγ receptors.
2Medicine,
065 ADIPONECTIN PROMOTES THE DIFFERENTIATION OF NAÏVE T CELL TO TH17 CELL AND
AGGRAVATES COLLAGEN-INDUCED ARTHRITIS
M. Zhang1, X. Sun1, X. Feng1, K. Gan1, W. Tan1
1Rheumatology
Department The First Affiliated Hospital of Nanjing Medical University, The First Affiliated Hospital of
Nanjing Medical University, Nanjing, China
 Background and aims:
To investigate a role of Adiponectin (AD) on the differentiation of na•ve T cell to Th17 cell and bone erosion in
collagen-induced arthritis (CIA) mice.
 Methods:
Purified na•ve T cells were cultured with TGF-β, IL-6, IL-23 in the presence and absence of AD (0.1, 1 or 10 μg/ml).
After 72 hours of culture, the frequencies of Th17 cells were measured by flow cytometry; the expression of ROR-γt
and IL-17 were determined by real-time PCR and ELISA. Intraarticularly injected of AD 10 μl (1 μg/1 μl) into the knee
joint of CIA mice was to analysis the role of AD on CIA development and Th17 expression in vivo.
 Results:
The frequencies of Th17 cells were significantly increased with the concentration of AD 10 μg/ml, but AD treatment
with 0.1 and 1 μg/ml showed no obvious effect on the differentiation of na•ve T cell to Th17 cell. The expression of IL17 and ROR-γt mRNA were significantly increased cupon AD 0.1 μg/ml and 10 μg/ml AD stimulation. Consistently
with mRNA expression, IL-17 levels were significantly elevated in culture supernatants after 10 μg/ml AD treatment.
Intraarticular injection of AD into the left knee joint of CIA mice aggravated arthritic development and bone
erosion,triggered higher expression of IL-17 mRNA and its transcriptiona factor including ROR-γt, IL-21, IL-22 and IL23 in CIA model.
 Conclusions:
These findings identify a role of AD on enhancing the differenation of na•ve T cell to Th17 cell, contributed to CIA
bone erosion in CIA mice.
066 CELLULAR- MOLECULAR RELATION OF SERUM IL10 LEVELS WITH HYPERALGESIA VARIATION ON
ARTHRITIS IN RAT
J. Zaringhalam Moghadam1, Z. Zeinab Akhtari2, A. Akram Eidi2
1Physiology,
Shahid Beheshti University medical Science, Tehran, Iran
Department of Biology Science and Research Branch Islamic Azad University, Tehran, Iran
Introduction: Regarding to the important anti-inflammatory role of IL10during inflammation process and hyperalgesia
and edema variation during CFA-induced arthritis and also the increase of Spinal mu opioid receptor (mOR)
expression, we aimed to investigate the role of serum IL10 level on mOR expression and edema and hyperalgesia
variation during different stages of Complete Freund`s Adjuvant (CFA) - induced arthritis in male Wistar rats.
Material & Method: Mono-arthritis was induced by CFA and inflammatory symptoms (hyperalgesia and edema) were
assessed on 0, 3,7th, 14th and 21st days of study. Anti-IL10was administered during the 21 days of study in different
experimental groups. mOR expression were detected by western blotting.
Results: Our results showed that anti-IL10 administration in AA (Adjuvant Arthritis) group caused an increase in the
paw volume and hyperalgesia until 21st of study. Our study stated that there were no significant differences in spinal
mOR expression between AA and AA+anti-IL10rats
Conclusion: Our study confirmed that anti-IL10administration caused to hyperalgesia and edema during AA
inflammation. Also these findings suggested that mOR expression increased in chronic phase of AA inflammation,
however an increase in the level of spinal mu opioid receptor (mOR) expression during AA inflammation is not
mediated directly via the effect of serum IL-10.
2Biology,
067 ARE AUTOANTIBODIES AGAINST BETA2-GLYCOPROTEIN I REALLY ATHEROGENIC?: ANSWERS
FROM IN VIVO PET IMAGING STUDIES IN APOLIPOPROTEIN E-DEFICIENT MICE
Y. Matsunami1, T. Sasaki2, F. Takenaka2, K. Kobayashi2, K. Kojima3, S. Kita3
1Department
of Cell Chemistry, Okayama University Graduate School of Medicine Dentistry and Pharmaceutical
Sciences, Okayama, Japan
2Collaborative Research Center for (OMIC), Okayama University Graduate School of Medicine Dentistry and
Pharmaceutical Sciences, Okayama, Japan
3IVD Development Department, Medical and Biological Laboratories Co Ltd, Ina, Japan
Background: β2-glycoprotein I (β2GPI) is a major autoantigen for antiphosholipid antibodies that appear in patients
with antiphospholipid syndrome (APS). Our in vitro studies have demonstrated that β2GPI specifically binds oxidized
low-density lipoprotein (oxLDL) to subsequently form pro-atherogenic/proatherothrombotic IgG immune complexes.
However, this proposed mechanism has not been confirmed up to this date in any type of in vivo studies.
Methods and Results: We reconstructed a newly established IgG murine antibody (3H3) specific for theβ2GPI
molecule when bound to oxLDL into its single chain Fv (3H3-scFv, 25 kDa) antibody variant lacking the Fc region.
Both 3H3-scFv and 3H3-IgG (as full sized IgG) revealed a quite similar specificity to anti-β2GPI autoantibodies
derived from APS patients and from the animal APS model NZW x BXSB Fl male mice by ELISA and
immunohistochemical analysis. The specific accumulation of 64Cu-labeled 3H3-scFv was observed in Sudan IVpositive atherosclerotic lesions of aortic valves and thoracic/abdominal arteries from high fat-loaded apolipoprotein Edeficient (ApoE-/-) mice injected intravenously 24 hours prior to PET imaging. The bio-distribution analyses also
confirmed the quantitative and significant accumulation.
Conclusion: The demonstration of 3H3-scFv accumulated in atherosclerotic lesion after its intravenous injection
represents a novel and significant observation to support the idea that APS derived autoantibodies can
immunologically target atherogenic oxLDL/β2GPI complexes in arterial lesions. This interaction results in IgG immune
complex formation that as previously demonstrated induce autoimmune-mediated atherosclerosis/and atherothrombosis.
Free Communication: Free Communications 1- Pearls in SLE, APS and RA research
069 CLINICAL EVALUATION OF TOTAL AND HIGH AVIDITY ANTI-DSDNA ANTIBODY EIA FOR THE
DIAGNOSIS OF SYSTEMIC LUPUS ERYTHEMATOSIS
H. Cao1, S. Zhou2, P. Wan1, W. Li1, T. Prestigiacomo2, J. Zheng1
1Ruijin
Hospital, Jiaotong University, Shanghai, China
Bio-Rad Laboratories, Hercules, USA
This study is to investigate clinical utility of total (high and low avidity) and high avidity (HA) anti-dsDNA antibodies in
an enzyme-linked immunosorbent assay (EIA). A total of 431 sera, 221 from SLE patients, 51 from patients with other
autoimmune diseases and 159 from healthy subjects, were measured using five different formats of anti-dsDNA EIA
kits. Two kits are commercially available at Bio-Rad Laboratories, the remaining are research use only kits. Two kits
detect total anti-dsDNA IgG antibodies, two kits measure HA anti-dsDNA IgG antibodies, and one detects anti-dsDNA
antibodies of both IgG and IgM isotypes. High avidity EIA tests have fewer false positives, especially in other
autoimmune disease control group compared to total anti-dsDNA assays (8/51 vs 1/51 and 6/51 vs 2/51). Although all
the assays were able to show a correlation of anti-dsDNA antibody with disease activity and low C3 and C4 levels
(P<0.0001), inactive SLE patients have more negatives on high avidity anti-dsDNA assays. The presence of antidsDNA antibody in SLE patients is associated with lupus nephritis and serositis. Mann Whitney and Fisher exact
analyses show there are significant differences of anti-dsDNA antibody presence and level in the patients with or
without kidney damage (P<0.0001), but not in the patients with or without hematological damage (P>0.05). The ratio
of HA to total anti-dsDNA antibodies is significantly higher in patients with M-SLEDAI >4 or with active kidney damage
(t test, P<0.0001), and it may be useful for monitoring SLE patients as it demonstrated a correlation to disease
activity.
2CDG,
070 ENDOGENOUS SLPI RELEASED BY RHEUMATOID SYNOVIAL FIBROBLASTS CONTROL BAFFDEPENDENT-B CELL ACTIVATION IN VITRO AND IN THE CIA AND RA/SCID-ARTHRITIS MODELS
N.W. Kam1, F. Brentano2, D. Kyburz2, S. Gay2, A. Filer3, C. Buckley3, C. Pitzalis1, M. Bombardieri1
1Experimental
Medicine and Rheumatology, William Harvey Research Institute, London, United Kingdom
of Rheumatology, University Hospital Zürich, Zürich, Switzerland
3Division of Immunity and Infection, College of Medical and Dental Sciences University of Birmingham, Birmingham,
United Kingdom
Background and aims: Secretory leukocyte protease inhibitor (SLPI) is a serine protease inhibitor with potent antimicrobicidal/immunoregulatory activities. Rheumatoid arthritis (RA) is characterized by synovial niches of
autoreactive B cells. Autocrine production of B cell survival factor BAFF by RA synovial fibroblasts (RASF) supports
ongoing B cell activation within the RA synovium. We investigated whether SLPI: (1)is produced by Toll-like receptors
(TLRs)-treated RASF and regulates B cell activation; (2)exerts immunoregulatory effects in the RA synovium/SCID
chimeric model and in collagen induced arthritis (CIA).
2Department
Methods: mRNA and protein expression of SLPI in RASF/RADF (dermal) stimulated with/without TLR2/TLR3/TLR4
ligands was assessed by QT-PCR and ELISA. RASF were treated with/without recombinant SLPI to study: (1)BAFF
expression; (2)AID expression and class-switching in co-culture with IgD+ B cells. BAFF expression and antibody
production were examined in rSLPI-treated RA/SCID mice. Severity of arthritis, anti-CII antibodies, and joint
histopathology were studied in rSLPI-treated CIA mice.
Results: Stimulation of RASF with TLR3-ligands led to a 15-fold induction of SLPI mRNA. SLPI protein was timedependently released from TLR3-stimulated RASF, but not RADF. SLPI restrained the production of BAFF, AID and
IgA/G/M in TLR3-treated RASF and co-cultures, respectively. SLPI reduced BAFF expression and IgG/IgM
production in RA/SCID mice while severity of arthritis, cartilage damage and anti-CII-IgG2a were reduced in SLPItreated CIA.
Conclusions: RASF release high levels of SLPI constitutively and upon TLR3 stimulation. SLPI directly modulates
BAFF and B cell activation in vitro/in vivo and reduces joint inflammation in CIA, highlighting a novel endogenous
anti-inflammatory pathway with therapeutic potential in RA.
071 RENAL INVOLVEMENT MORE FREQUENT AND MAY BE LINKED TO POORER SURVIVAL AMONG
FILIPINO MALES WITH SLE
M.A.C. Saquing1, S. Tupas1, S.V. Navarra1
1Medicine
Section of Rheumatology, University of Santo Tomas, Metro Manila, Philippines
Objective: To compare the organ involvement and survival of Filipino males vs. females with systemic lupus
erythematosus (SLE).
Methods: Retrospective study describing the survival rate of Filipino male lupus patients seen at the Rheumatology
Clinics of the University of Santo Tomas (UST) Hospital, Manila, Philippines. Of the 1470 patients entered in the UST
Lupus Database from 1998 - 2012, 95 (6.46%) were males. Age at SLE diagnosis, median survival rate , and causes
of mortality were described.
Results: Average age at diagnosis of males was 27.33 ± 12.6 years. Average disease duration was 63 ± 57 months
vs. females at 99 ± 75.66 months. In males, renal involvement (58% vs 41% , p<0.0008) and hematologic
manifestations (55% vs 42% , p< 0.0139) were more common, whereas arthritis (58% vs 71%, p= 0.0152) and
photosensitivity (38% vs 52% p= 0.0150) were significantly less frequent. Median survival rate of males was 4 years
vs 7 years for females. Of the 22 male (23%) who died , the average disease duration was 48 + 46 with renal failure
(27%) and infection (27%) as the primary causes of death.
Conclusion: Renal and hematologic manifestations were more common in males than females, whereas the more
symptomatic musculoskeletal and cutaneous ie photosensitivity manifestations were more common in females *
reinforcing the need for earlier detection and aggressive management of renal involvement especially in males with
SLE. Poorer survival also appeared to be linked with renal involvement in this group of Filipino male SLE patients.
072 NATURAL KILLER AND NATURAL KILL T CELL DYSFUNCTION IN ADULT-ONSET STILL'S DISEASE
Y.W. Park1, Y.N. Cho1, T.J. Kim1, H.M. Jin1, H.J. Jung1, J.Y. Kim1, M.J. Kim1, S.J. Kee2
1Rheumatology,
Chonnam National University Hospital, Gwangju, Korea
Medicine, Chonnam National University Hospital, Gwangju, Korea
Aims. To examine the levels and functions of natural killer (NK) and natural killer T (NKT) cells, investigate
relationships between NK and NKT cells, and determine the clinical relevance of NKT cell levels in patients with
adult-onset Still's disease (AOSD). Methods. Patients with active untreated AOSD (n=20) and age- and sex-matched
healthy controls (n=20) were studied. NK and NKT cell levels were measured by flow cytometry. Peripheral blood
mononulclear cells were cultured in vitro with α-galactosylceramide (α-GalCer). NK cytotoxicity against K562 cells
and proliferation indices of NKT cells were estimated by flow cytometry. Results. Percentages and absolute numbers
of NKT cells were significantly lower in the peripheral blood of AOSD patients than in that of healthy controls.
Proliferative responses of NKT cells to α-GalCer were also lower in patients, and this was found to be due to
proinflammatory cytokines and NKT cell apoptosis. In addition, NK cytotoxicity was found to be significantly lower in
patients than in healthy controls, but NK cell levels were comparable in the two groups. Notably, this NKT cell
deficiency was found to be correlated with NK cell dysfunction and to reflect active disease status. Furthemore, αGalCer-medicated NK cytotoxicity, showeing the interaction between NK and NKT cells, was signficantly lower in
AOSD patients than in healthy controls. Conclusions. These findings demonstrate that NK and NKT cell functions are
defective in AOSD patients and suggest that these abnormalities contribute to innate immune dysfunction in AOSD
2Laboratory
073 LOW SERUM LEVEL OF 25-HYDROXYVITAMIN D3 IN NEWLY ONSET SYSTEMIC LUPUS
ERYTHEMATOSUS PATIENTS: INTRINSIC ABNORMALITY?
X. Zhang1, O. Jin1, Q. Wei1, H. Huang1, C. Hou1, Z. Lin1, Z. Liao1, D. Lin1, Q. Li1, J. Gu1
1Rheumatology,
The Affiliated Third Hospital of Sun Yat-san University Rheumatology, Guangzhou, China
OBJECTIVES:
Vitamin D deficiency has been found in patients with SLE, however, whether this deficiency is due to SLE intrinsic
abnormality or a result of some other reasons (such as avoidance of sunshine, photoprotection, renal insufficiency
and the use of medications including glucocorticoids and HCQ) are still not clear. We aim to investigate the serum
25(OH)D levels on newly diagnosed SLE patients, further more, we want to evaluate the relationship between
25(OH)D level and the bone degradation marker (amino-terminal cross-linked telopeptide of type I collagen, ICTP)
and SLE disease activity index (SLEDAI).
METHODS:
SLE patients were divided into 3 groups: (1) newly onset SLE (2) avtive SLE (3) stable SLE. Age and sex matched
healthy donor were recruited. Both group 2 and 3 patients were given Vitamin D and Calcium to avoid GIOP. Serum
levels of 25(OH)D and ICTP were measure by ELISA.
RESULTS: (1) The level of 25-vitamin D in newly on-set SLE patients (32.75±3.20 nmol/L) was significantly lower
than active (60.50±7.21 nmol/L) or stable (59.23±5.06 nmol/L) SLE patients and healthy donors (60.97±4.89 nmol/L)
(P<0.05, respectively), while there were no differences between active, stable SLE patients and the healthy donors.
No correlation was found between the level of 25-vitamin D and ICTP, or level of 25-vitamin D and SLEDAI, in
healthy donors, newly on-set, active and stable SLE patients respectively. (2) The level of ICTP in newly on-set SLE
patients (13.05±3.00 ug/L) and active SLE (9.23±2.59 ug/L) was significantly higher than stable SLE patients
(5.13±1.14 ug/L, p=0.009). The levels of ICTP were respectively correlated with SLEDAI (newly on-set SLE: (r=0.70,
p=0.016), active SLE: (r=0.83, p=0.001) and stable SLE: (r=0.59, p=0.005)).
CONCLUSIONS:
1. The low level of 25-vitamin D in newly on-set SLE patients suggests that this may be an intrinsic abnormality,
whose regulation to immune system is insufficient, which may contribute to the pathogenesis of SLE. After treat with
Vitamin D and Calcium, both active or stable SLE reached normal 25-vitamin D level.
2. The level of ICTP in newly on-set SLE patients and active SLE patient was significantly higher than stable SLE
patients, and was correlated with SLEDAI,suggesting that the inflammation of SLE may influence the bone
degradation.
074 MATERNAL AND FETAL OUTCOMES OF LUPUS PREGNANCIES AMONG FILIPINO PATIENTS IN A
TERTIARY HOSPITAL
C. Tan1, L.R.L. Lizardo1, S.V. Navarra1
1Rheumatology,
University of Santo Tomas Hospital, Manila, Philippines
Background and Objective: Pregnancy poses a relevant management challenge in systemic lupus erythematosus
(SLE), a disease which usually affects women of child-bearing age. Maternal and fetal outcomes were retrospectively
reviewed in this study. Design: SLE patients who became pregnant after diagnosis were included. Results: There
were 85 patients with 184 pregnancies occurring during the course of the disease. The mean age in years at SLE
diagnosis was 26.19 while the mean age in years at pregnancy was 28.74. Majority delivered via cesarean section
(54.7%) with 92.3% of infants having a normal birth weight. Pre-pregnancy co-morbidities include chronic
hypertension (6.5%), Tuberculosis (TB) (2.7%) and Lupus Nephritis (LN) (9.2%). During pregnancy, maternal
complications were gestational diabetes (GDM) (0.54%), pre-eclampsia (8.2%), varicella-zoster (VZV) infection
(1.6%) and hyperthyroidism (0.54%). Post-pregnancy complications include pericardial effusion (0.54%), pericarditis
(0.54%), dilated cardiomyopathy (0.54%), post partum depression (0.54%), and hypertension (0.54%). Among the
pregnancy outcomes, term deliveries were recorded in 97 (82.9%) patients while 20 (17.1%) had preterm deliveries.
There were 67 (36.4%) nonviable pregnancies manifested as miscarriages (83.6%), blighted ovum (8.9%) and
intrauterine fetal demise (IUFD) (7.5%). Congenital diseases noted after delivery include congenital heart block
(1.7%), meningocoele (0.86%), thyroid abnormality (0.86%), G6PD (0.86%), neonatal lupus (0.86%) and autism
(0.86%). Conclusion: Although successful pregnancy outcomes are possible for SLE patients, miscarriages, preterm
deliveries, blighted ovum and IUFD remain a concern, requiring close monitoring and effective multi-specialty team
approach.
Funding: Lupus-Inspired Advocacy Project of Rheumatology Educational Trust Foundation Inc.
075 RECOMBINANT DOMAIN V OF BETA2-GLYCOPROTEIN I INHIBITS THE FORMATION OF ATHEROGENIC
OXLDL/BETA2-GLYCOPROTEIN I COMPLEXES
Q.P. Liu1, Y. Chi1, J.D. Li1, R.J. Wang1, E. Matsuura2
1College
of Life Science and Technology, Dalian University, Dalian, China
of Cell Chemistry, Okayama University Graduate School of Medicine Dentistry and Pharmaceutical
Sciences, Okayama, Japan
Background: Oxidized low-density lipoproteins (oxLDL) interact with beta2-glycoprotein I (β2-GPI) to form oxLDL/β2GPI complexes implicated as autoantigens that promote the development of atherothrombotic vascular complications
in patients with antiphospholipid syndrome (APS). The region of β2-GPI responsible for the binding with oxLDL has
been located in the same phospholipid-binding region of domain V (DV). Anti-oxLDL/β2-GPI antibodies enhance the
in vitro macrophage uptake of oxLDL that explains the accelerated progression of autoimmune-mediated
atherothrombosis. We hypothesized that DV of β2-GPI may competitively inhibit the formation of circulating
oxLDL/β2-GPI complexes.
Methods and results: DV was expressed in the Pichia pastoris (P. pastoris) (P.rβ2-GPI DV) expression system, and
used in binding assays. Enzyme-linked immunoassay (ELISA) demonstrated that P.rβ2-GPI DV interacted with
oxLDL via 7-keocholestryl-9-carboxynonanoate (oxLig-1), a negatively charged lipid moiety from oxLDL. It was also
shown that free ω-carboxyl residue of oxLig-1 was required for this interaction. Serologic tests also showed elevated
levels of oxLDL/β2-GPI complexes in patients with APS. Moreover, P.rβ2-GPI DV bound oxLDL with a high affinity in
APS sera, and that the addition of P.rβ2-GPI DV inhibited the formation of circulating oxLDL/β2-GPI complexes.
Conclusions: Recombinant P.rβ2-GPI DV was capable of binding to oxLDL, and this binding could effectively inhibit
the formation of oxLDL/β2-GPI complexes in the sera of the APS patients, suggesting that P.rβ2-GPI DV could
potentially be a new candidate compound for autoimmune disease therapy.
2Department
Free Communication: Free Communications 2- Pathogenetic and Immunoregulatory Mechanisms in
Autoimmunity
077 ROLES OF TOLL-LIKE RECEPTORS IN GENE/ENVIRONMENT INTERACTIONS IN A MOUSE MODEL OF
MULTIPLE SCLEROSIS
S. Miranda-Hernandez1, S. Chowdhury1, M. Jordan1, A.G. Baxter1
1Comparative
Genomics Centre, James Cook University, Townsville, Australia
The most commonly used animal model for the study of Multiple Sclerosis (MS) is Experimental Autoimmune
Encephalomyelitis (EAE). EAE causes a demyelinating paralysis resembling some forms of Multiple sclerosis (MS).
Previously we found that C57BL/6.Tlr2-/- mice decrease the severity of EAE. In addition, transfer of EAE into
C57BL/6.Tlr2-/- mice failed to transfer disease. This protection from EAE was associated with fewer infiltrating CD4 + T
cells in the CNS, reduced prevalence of detectable circulating IL-6, and increased proportions of central (CD62L+)
Foxp3+ Tregs. As these results indicate that TLR signalling affects CNS-autoimmunity, in this study we examined the
role of TLRs in NOD mice, which are susceptible to Type 1 Diabetes (T1D) in order to dissect the interrelationships
between organ-specific autoimmune diseases.
NOD/Lt mice develop a mild form of relapsing/remitting EAE. In contrast to C57BL/6 mice, TLR2 deletion in NOD
mice did not affect the severity of disease. We tested whether insulitis associated with T1D development
compensated for the lack of TLR2 signalling in these mice. The course of EAE was analyzed in NOD.H2b and
NOD.H2d mice, which do not develop insulitis or T1D. NOD.H2b mice showed increased severity of EAE compared to
NOD/Lt mice, again in a relapsing/remitting pattern. In the absence of insulitis, EAE was ameliorated in
NOD.H2b.Tlr2-/- mice and no relapses occurred after the initial peak of disease. These data suggest that both TLR2
signalling and immunological events associated with autoimmunity at a distant site can mediate relapses in CNS
autoimmunity.
078 POSTTRANSCRIPTIONAL GENE REGULATION OF CD4+ TH17 DIFFERENTIATION BY THE RNA-BINDING
PROTEIN HUR
U. Atasoy1, J. Chen1, J. Cascio1, J.D. Magee1, P. Techasintana1, M.M. Gubin1, G.M. Dahm1, R. Calaluce1
1Surgery,
University of Missouri, Columbia, USA
Interleukin 17 (IL-17A) is a proinflammatory cytokine produced by activated CD4+ Th17. Th17 cells are major
contributors to autoimmune diseases, including multiple sclerosis. Although the transcriptional control of IL-17 and
Th17 differentiation have been well studied, posttranscriptional gene regulation is unclear. The RNA-binding protein
(RBP) HuR regulates the stability of many target mRNAs via binding the AU-rich elements (ARE) present in the 3Õ
untranslated region (UTR), of many pro-inflammatory cytokines such as IL-4, IL-13 and TNF. Since IL-17 mRNA also
possesses HuR binding motifs, we investigated whether HuR can regulate Il-17 expression. We demonstrated that
HuR directly binds IL-17 mRNA 3Õ UTR by using RNA immunoprecipitation methods as well as biotin pull down
assays. CD4+ T cells from HuRfl/fl KO mice have decreases in Th17 differentiation, IL-17 steady-state mRNA and
protein expression compared to wild type Th17 cells. IL-17 mRNA stability was decreased by 3-fold in HuR KO mice.
We used the experimental autoimmune encephalitis (EAE) model of neuroinflammation to investigate whether HuR
plays a role in CD4+ Th17 differentiation and disease initiation. Mice with adoptive transfer of HuR KO CD4 + T cells
had delayed onset, reduced severity of EAE symptoms, as well as reduced CNS inflammation and decreased IL-17
production by Th17-producing cells recovered from the CNS. Our results reveal HuR-controlled posttranscriptional
regulatory mechanisms of Th17 differentiation and direct effects upon EAE initiation. These findings could provide
novel therapeutic targets for treatment of IL-17- mediated autoimmune neuroinflammation and also potentially other
diseases in which IL-17 plays a major role.
079 EXPRESSION OF INFLAMMASOME IS DIFFERENT IN MUSCLE OF DERMATOMYOSITIS AND
POLYMYOSITIS PATIENTS
Q.Z. Zhao1, Y.L. Zhang1, H. Zhou1, X.M. Shu1, X. Lu1, G.C. Wang1
1Rheumatology,
China-Japan Friendship Hospital, Beijing, China
Background and aims Activation of innate immune plays important role in the pathogenesis of autoimmune disease.
Our study was to investigate the expression of inflammasomes in muscle of dermatomyositis(DM) and
polymyositis(PM) patients.
Methods Immunohistochemistry was performed to determine the expression of NALP (NACHT-LRR-PYD-containing
protein)1, NALP3, ASC (apoptosis-associated speck-like protein containing a CARD) and caspase-1 in muscle of 10
newly diagnosed and untreated adult DM/PM patients (4 DM and 6 PM) and 5 healthy controls.
Results In muscle of 4 DM patients, ASC and caspase-1 were expressed in the plasma of CD3+ T and CD20+ B
cells, while in 2 of them NALP3 was also expressed. Only 1 PM patient in 6 expressed NALP3, ASC and caspase-1
in the plasma of CD3+ T and CD20+ B cells. NALP1 was not expressed in all of the 10 patients. No NALP1, NALP3,
ASC and caspase-1 were expressed in the muscle of 5 healthy controls. The positive rate of ASC and caspase-1 in
DM group was significantly higher than that in PM and control group (P<0.01 and P<0.05), while the positive rate of
NALP3 in DM group had no difference with that in PM and control group. The level of creatine kinase, erythrocyte
sedimentation rate, C-reactive protein had no difference between NALP3, ASC and caspase-1 positive group and
negative group.
Conclusions Innate immune plays different role in the pathogenesis of DM and PM. Inflammasome participates in
the inflammation of DM but not in PM, and may be activated by NALP3.
080 CD98HC REGULATES THE DEVELOPMENT OF EXPERIMENTAL COLITIS BY DIFFERENTIALLY
CONTROLLING INDUCIBLE AND NATURALLY OCCURING REGULATORY T CELLS
Z. Bhuyan1, H. Arimochi1, K. Yasutomo1
1Immunology
and Parasitology, Univ of Tokushima Grad Sch of Health Biosciences, Tokushima, Japan
CD4+ T cell activation is controlled by signaling through the T cell receptor in addition to various coreceptors, and is also affected by their interactions with effector and regulatory T cells in the
microenvironment. Inflammatory bowel diseases (IBD) are caused by the persistent activation and expansion
of auto-aggressive CD4+ T cells that attack intestinal epithelial cells. However, the molecular basis for the
persistent activation of CD4+ T cells in IBD remains unclear. In this study, we investigated how the CD98
heavy chain (CD98hc, Slc3a2) affected the development of colitis in an experimental animal model.
Transferring CD98hc-deficient CD4+CD25- T cells into Rag2-/- mice skewed CD4+ T cells toward Foxp3+
inducible regulatory T cells (iTregs), rather than their differentiation into Th1 or Th17 cells, and did not
induce colitis. By comparison, CD98hc-deficient naturally occurring regulatory T cells (nTregs) had a
decreased capability to suppress colitis induced by CD4 +CD25- T cells, although CD98hc-deficient mice did
not have a defect in the development of nTregs. Blocking CD98hc with an anti-CD98 blocking antibody
prevented the development of colitis. Our results suggest that CD98hc has distinct roles in iTregs and
nTregs in vivo during the progression of colitis and also suggest that CD98hc on CD4+ T cells contributes to
immune dysregulation in autoimmune colitis.
081 C-FLIP OVEREXPRESSION IS ASSOCIATED WITH PARANEOPLASTIC MYASTHENIA GRAVIS (MG) AND
LATE-ONSET MG (LOMG)
D. Belharazem1, D. Belharazem1, A. Grass1, B. Schalke2, A. Marx1
1Pathology,
University Hospital Mannheim, Mannheim, Germany
University Hospital Regensburg, Mannheim, Germany
Tumors of the thymus especially lymphoepithelial thymomas are 30%associated with the autoimmune Myasthenia
gravis (MG). The auto reactive T lymphocytes generated in thymus tumors stimulate antibodies production against
several muscle antigens, thereby acquiring a heterogeneous neuromuscular junction disease characterized by
muscular weakness.
Antiapoptotic proteins overexpression is one of the tumor-promoting mechanism. We ask whether the antiapoptotic
molecule cFlip may be operative in thymoma and thymoma associated MG and in LOMG.
2Neurology,
We investigated 87 thymomas (10A, 20AB, 20B2. 15B3 and 22 thymus carcinomas (TC)) without MG, 21 thymomas
associated MG(8AB, 8B2 and 5B3) and 13 thymic hyperplasia from late-onset MG (LOMG) for the expression of cFlip
using real time RT-PCR and western blotting for cryo-preserved material and immunohistochemistry (IHC) for
formalin-fixed, paraffin-embedded tissues. cFlip expression was also performed in peripheral blood lymphocytes from
LOMG, thymomas associated and not associated MG and compared to lymphocytes from donors.
An overexpression was detected in all thymomas tissues without MG and in thymus carcinomas (p<0.0001****). In
paraneoplastic MG cFlip mRNA and proteins Level were higher expressed (p<0.0001****).The non paraneoplastic
MG cases showed also a significant cFlip mRNA transcription compared to normal thymi. In addition cFlip was highly
detectable in paraneoplastic and non paraneoplastic MG Lymphocytes.
The anti-apoptotic protein cFlip is high expressed on the transcriptional and proteomic level in thymomas especially
when they are associated with MG. This suggests that cFlip might play a role in the pathogenesis of paraneoplastic
MG deserves future studies for its regulation as a new therapeutic target.
082 TYROSINE KINASE BTK IS REQUIRED FOR NK CELL-MEDIATED AUTOIMMUNE HEPATITIS
Y. Bao1, J. Zheng2, C. Han3
1Translational
Medicine Center, Shanghai Changzheng Hospital Second Military Medical University, Shanghai, China
of Paediatrics and Adolescent Medicine, Li Ka Shing Faculty of Medicine the University of Hong Kong,
Hong Kong, China
3National Key Laboratory of Medical Immunology & Institute of Immunology, Second Military Medical University,
Shanghai, China
BrutonÕs tyrosine kinase (Btk) is not only critical for B cell development and differentiation, but also involved in the
regulation of Toll-like receptor (TLR)-triggered innate response of macrophages. However, whether Btk is involved in
the regulation of NK cell innate function remains unknown.
Through FACS and immunoblot, we show that Btk expression is upregulated during maturation and activation of
mouse NK cells. Murine Btk-/- NK cells have decreased innate immune responses to TLR3 ligand, with reduced
expressions of IFN-γ, perforin and Granzyme-B, and decreased cytotoxic activity. Furthermore, Btk is found to
promote TLR3-triggered NK cell activation by activating NF-κB pathway. Poly I:C-induced, NK cell-mediated acute
hepatitis is observed to be attenuated in Btk-/- mice or the mice with in vivo administration of Btk inhibitor. Accordingly,
liver damage is aggravated in Btk-/- mice once adoptive transfer of Btk+/+ NK cells, further indicating that Btk-mediated
NK cell activation contributes to TLR3-triggered acute liver injury. Importantly, reduced TLR3-triggered activation of
human NK cells is observed in Btk-deficient patients with X-linked agammaglobulinemia (XLA), as evidenced by the
reduced IFN-γ, CD69, CD107a expression and cytotoxic activity.
These results indicate that Btk is required to activation of NK cells, thus providing insight into the physiological
significance of Btk in the regulation of immune cell functions and innate inflammatory response.
2Department
083 NEURO-VASCULAR DISRUPTION IN THE CEREBRAL CORTEX DURING EXPERIMENTAL AUTOIMMUNE
ENCEPHALOMYELITIS AND THE EFFECT OF GLATIRAMER ACETATE
R. Aharoni1, R. Eilam2, V. Kalchenko2, Y. Kuznetsov2, M. Sela1, R. Arnon1
1Immunology,
The Weizmann Institute of Science, Rehovot, Israel
Resorces, The Weizmann Institute of Science, Rehovot, Israel
Blood brain barrier (BBB) dysfunction is a key pathological event in multiple sclerosis (MS) and its model
experimental autoimmune encephalomyelitis (EAE). The cerebral cortex is an important target for the disease
process, but its pathophysiology is not yet understood. In this study we analyzed the function of the BBB and the
communication between the blood vessels, the astrocytes and the nerves in the cortex of MOG-induced EAE inflicted
mice. The consequences of treatment by glatiramer acetate (GA, Copaxone) on these processes were also
investigated.
We developed a system in which permeabilization of sodium fluorescein into the CNS indicated BBB functionality.
Extensive sodium fluorescein leakage was observed in EAE-induced mice, but not in na•ve controls. Notably, BBB
disruption was evident even before the development of clinical manifestations. In mice treated by GA either at
disease induction (prevention) or after disease exacerbation (therapeutic treatment), sodium fluorescein leakage was
significantly lower. Immunohistochemical analysis of blood vessels, astrocytes and neurons revealed loss of
astrocytes processes towards the blood vessels as well as the neurons in the cortex of EAE-inflicted mice. GA
treatment significantly reduced these manifestations resulting in normal appearing neurovascular formation. These
findings support the central role of BBB disruption and vascular pathology in the disease, suggesting that
neurovascular changes lead to abstracted crosstalk between the blood vessels and the neurons. Additionally, we
herein demonstrate a new therapeutic consequence of GA treatment. Specifically, GA treatment prevents the leakage
of BBB and reduces the neurovascular damage.
2Veterinary
084 MYCOPLASMA SUPERANTIGEN AS A TRIGGER PROMOTES AUTOREACTIVE T CELL RESPONSES AND
DISEASE SEVERITY IN AUTOIMMUNE DIABETIC MOUSE
H. Mu1
1Internal
Medicine, University of Utah School of Medicine, Salt Lake City, USA
Background and aims: Infection has been extensively documented as a serious problem causing high mortality and
morbidity in diabetic populations. Due to the complex nature of immunological mechanisms, little is known regarding
how an infection affects the outcomes of diabetes. Our laboratory has previously established a mouse model of septic
shock by infecting mice with the Mycoplasma arthriditis. M. arthritidis produces a potent superantigen (SAg) MAM,
which has been shown to be at least partially responsible for the lethal toxicity induced in certain susceptible mouse
strains.
Methods: In the present study, we have investigated how diabetic immune cells respond to the M. arthritidis and MAM
SAg. By using two well-defined animal models of diabetes, STZ and NOD mice.
Results: We found that STZ mice infected with M. arthritidis developped severe lethal toxicity and the immune cells
predominantly produces pro-inflammatory cytokines TNF?, IL-6, IL-1β, chemokines CCR2/4 and Tbx21, a Th1related transcription factor; while MAM-activated T cells from NOD mice display a mixture of Th1 and Th17 profiles
which are associated with the elevated levels of ICOS molecule and a unique transcription factor LSF (tcfcp2). Since
ERK, a signaling protein of MAPK family, is reported to be responsible for the phosphorylation of LSF in T cell
activation, thus we propose a novel ERK-LSF-ICOS signaling cascade that is required for the Th1/Th17 development
in NOD mice in response to MAM.
Conclusions: Our findings suggest that the inhibition of these molecules might be potentially therapeutic in the
alleviation of mortality and morbidity in diabetic individuals with severe infections.
This work was supported by a grant from the Nora Eccles Treadwell Foundation
085 UP-REGULATION OF MICRORNA-210 INDUCES IMMUNE DYSFUNCTION VIA TARGETING FOXP3 IN
CD4+ T CELLS OF PSORIASIS VULGARIS
L.T. Wang1, Y.W. Su1, G.P. Liang1, H.Y. Zhai1, P. Zhang1, X.J. Deng1, M. Zhao1, Q.J. Lu1
1Department
of Dermatology, Second Xiangya Hospital Central South University, Changsha, China
Background and aims Psoriasis vulgaris (PV) has been known as one kind of autoimmune skin diseases. The
mechanism of aberrant immunological regulation in PV remains unclear. Here, we investigated the role and
mechanism of microRNA-210 (miR-210) in CD4+ T cells of PV.
Methods. 18 PV patients and 18 healthy individuals were recruited and CD4+ T cells were isolated by magnetic
beads. The transcription levels of miR-210, FOXP3 and cytokines were detected by real-time PCR. Luciferase
reporter gene assay was used to identify the target gene of miR-210. CD4+ T cells were transfected by nucleofector
device in combination with the human T-cell nucleofector kit. FOXP3 protein level was detected by western blot. The
levels of cytokines in the supernatant of cells culture were measured using enzyme-linked immunosorbent assay.
Results The expression of miR-210 is up-regulated significantly in the CD4+ T cells of PV patients compared with
healthy controls, which are inversely correlated with the mRNA levels of FOXP3. Luciferase reporter gene assay
showed that FOXP3 is a target gene of miR-210. Transfection of miR-210 mimic leads to the reduced FOXP3, IL-10
and TGF-β expression and the increased IFN-γ expression in normal CD4+ T cells. In contrast, transfection of miR210 inhibitor increases FOXP3, IL-10 and TGF-β expression and reduces IFN-γ expression in CD4+ T cells of PV
patients.
Conclusions Our data demonstrates that up-regulation of miR-210 leads to immune dysfunction by targeting FOXP3
in CD4+ T cells of PV paitents, which suggests the important role of miR-210 in the pathogenesis of PV.
086 INTERLEUKIN 27 INHIBITS ATHEROSCLEROSIS VIA IMMUNOREGULATION OF MACROPHAGES IN MICE
H. Yoshida1, T. Hirase2, H. Hara1, K. Node3
1Department
of Biomolecular Sciences Division of Immunology, Saga University Faculty of Medicine, Saga, Japan
of Bioscience, National Cardiovascular Center Research Institute, Suita, Japan
3Department of Cardiovascular and Renal Medicine, Saga University Faculty of Medicine, Saga, Japan
Chronic inflammation in arterial wall that is driven by immune cells and cytokines plays pivotal roles in the
development of atherosclerosis. Interleukin 27 (IL-27) is a member of IL-12 family cytokines that is consisted of IL27p28 and EBI-3 and has anti-inflammatory properties that regulate T cell polarization and cytokine production. In this
study, the role of IL-27/IL-27 receptor signaling in the pathogenesis of atherosclerosis.
IL-27-deficient (Ldlr-/-Ebi-3-/-) and IL-27 receptor-deficient (Ldlr-/-WSX-1-/-) Ldlr-/- mice were generated and fed with a
high-cholesterol diet (high fat diet; HFD) to induce atherosclerosis. Roles of bone marrow-derived cells in vivo and
macrophages in vitro were studied using bone marrow reconstitution by transplantation and cultured peritoneal
macrophages, respectively.
We demonstrate that mice lacking IL-27 or IL-27 receptor are more susceptible to atherosclerosis compared to wild
type due to enhanced accumulation and activation of macrophages in arterial walls (See figure below.)
2Department
The number of circulating pro-inflammatory Ly6Chi monocytes showed no significant difference between wild type
mice and mice lacking IL-27 or IL-27 receptor. Administration of IL-27 suppressed the development of atherosclerosis
in vivo and macrophage activation in vitro that was indicated by increased uptake of modified low density lipoprotein
and augmented production of proinflammatory cytokines.
These findings define a novel inhibitory role for IL-27 in atherosclerosis that regulates macrophage activation in mice.
087 B-LYMPHOCYTE SUBSETS IN CHRONIC FATIGUE SYNDROME
K. Fuller1, E.W. Brenu1, T.K. Huth1, S.L. Hardcastle1, S. Johnston1, D.R. Staines2, S. Marshall-Gradisnik1
1National
Center for Neuroimmunology and Emerging Diseases, Griffith University, Southport, Australia
Coast Public Health Unit, Queensland Health, Robina, Australia
Background and Aims: Chronic Fatigue Syndrome (CFS) is a multi-factorial disease with a significant immune
component. To date, limited studies have focused on B-lymphocytes and their implications in CFS. However, it has
recently been shown that B-lymphocyte depletion therapy via the administration of Rituximab may be beneficial to
some CFS patients, thus suggesting a potential role of B-lymphocytes in CFS. Hence, this study aims to examine the
levels of specific B-lymphocyte phenotypes in CFS patients in comparison to non-fatigued control participants.
2Gold
Methods: Heparinised peripheral blood samples were collected from 38 CFS patients fulfilling the 1994 Centers for
Disease Control and Prevention criteria for CFS, and 29 non-fatigued control participants. B-lymphocyte phenotypes
were characterised using specific antibody combinations and analysed using flow cytometry protocols.
Results: The results showed significant differential numbers in B-lymphocyte subsets in CFS patients compared to
non-fatigued controls. Specifically, CFS patients exhibited decreased levels of immature B-lymphocytes, and
increased levels of memory B-lymphocytes when compared to those of non-fatigued control participants.
Conclusion: Results from the current study confirm a potential involvement of B-lymphocytes in CFS. B lymphocytes
may be an important immunological marker for the diagnosis of CFS. Hence, further studies are required to confirm
whether alterations in B-cell phenotypes are involved in the pathophysiology of CFS.
088 DISTURBANCES IN THE PHENOTYPIC AND FUNCTIONAL CHARACTERISTICS OF MEMORY T CELLS IN
PATIENTS WITH RHEUMATOID ARTHRITIS.
A. Khanniche1, P.H.D. Jiang Bin1
1School
of medecine, Shanghai JiaoTong University, Shanghai, China
Memory T cells afford heightened protective immunity against subsequent re-infection. It was shown that excessive
inflammation during chronic autoimmune diseases (eg: Rheumatoid arthritis) can have detrimental effects on the
generation of functional memory T cells. However, this effect is not yet well understood.
Our aim is:
- To show the impairment of memory T cells in different groups of RA patients through identification of memory CD4+
and CD8+ T cells phenotype and function (frequencies of IFN-g, TNF-a, IL-2, IL-17 positive cells)
Methods: Upon isolation of PBMC from peripheral blood, we proceeded to intracellular staining and flow cytometry.
Results:
We have shown an inhibitory profile in patients with established RA characterized by lower frequencies of memory
Th1 cells and TNF-a+CD8+ cells. Also, patients receiving DMARD alone or combined with Etanercept had fewer
functional memory CD4+ T cells and CD8+ T cells; lower frequencies of Th1 cells were observed in both groups
when compared to healthy individuals, as well as fewer IFN-g+CD8+CD45RA-T cells, fewer TNF- a +CD8+CD45RAT cells and fewer IL-2+ CD8+CD45RA- T cells were observed in those patients.
Furhtermore, we have demonstrated an imbalance in TEM/TCM frequencies characterized by a significant decrease
in TCM in RA patients, as compared to HC. This imbalance was observed mainly in patients under Golimumab a
TNF-? inhibitor and Tasocitinib a JAK inhibitor.
Conclusion: These results suggest that in a chronic inflammatory environment such as in patients with RA, the
homeostasis of memory T cells can be disrupted and may influence their response to subsequent infections.
Parallel Session: Parallel Session 5: Biosimilars &amp; related therapies
097 THE RISE OF BIOSIMILAR MABS
S.S. Hong1
1R&D,
Celltrion, Incheon, Korea
Celltrion is a global biopharmaceutical company, equipped with FDA-certified GMP production facility. Celltrion aims
to develop biosimilar products based on EMA regulations, and conduct global clinical trials to show comparability with
reference drugs. Celltrion has eight biosimilar candidates; some of those are Infliximab, Rituximab, Etanercept, and
Adalimumab. Currently, Infliximab Biosimilar has been approved by some countries including EU. Clinical trial phase I
for Rituximab has just finished and phase III will be starting soon including US.
The EMA published its guidelines for biosimilars of complex biologics in early 2012 1, and the US FDA has also
recently provided its draft guidelines for biosimilar approval.2 In response, many biopharmaceutical companies have
ramped up their efforts to gain marketing approval for biosimilars, especially for mAb products. A series of recent
journals and articles have demonstrated pipeline for development of biosimilars in autoimmune diseases. 3-4
Monoclonal antibodies are structurally very complex, rigorous non-clinical testing has performed to confirm quality,
structural and functional comparability in vitro in accordance of EMA guidance. Especially, RemsimaTM, the infliximab
biosimilar is the first monoclonal antibody biosimilar got approval from EMA. In May 2013, some of the clinical data
has been published along with 30 week data of phase I and III in supplementary of ARD journals.5-6 Celltrion will
supply high quality product consistently to provide better treatment option for those who need such benefits.
098 RANDOMIZED CLINICAL TRIALS TO DEMONSTRATE THE EQUIVALENCE OF CT-P13 WITH IINFLIXIMAB
IN PATIENTS WITH ANKYLOSING SPONDYLITIS AND RHEUMATOID ARTHRITIS
D.H. Yoo1
1department
of medicine, Hanyang University Hospital for Rheumatic Diseases, Seoul, Korea
Objectives: To demonstrate equivalence of CT-P13 to innovator infliximab in patients with ankylosing spondylitis in
PLANETAS trial and with rheumatoid arthritis in PLANETRA trial.
Methods: Patients were randomised to receive 5 mg/kg of CT-P13 (n=125) or INX (n=125) in PLANETAS trial;
3mg/kg of CT-P13 (n=302) or INX (n=304) with MTX and folic acid in PLANETRA trial at week 0, 2, 6 and then every
8 weeks up to week 54.
Results: In the PLANETAS trial, geometric mean AUCτ at steady state was 32,765.8μgh/mL for CT-P13 and
31,359.3μgh/mL for INX. Geometric mean Cmax,ss was 153.5μg/mL for CT-P13 and 150.4μg/mL for INX. ASAS20
and ASAS40 responses at week 30 were 70.5% and 51.8% for CT-P13 and 72.4% and 47.4% for INX, respectively.
Comparable efficacy profile between CT-P13 and INX maintained up to week 54.
In the PLANETRA trial, ACR20 were 60.9% for CT-P13 and 58.6% for INX (95% CI: -6%, 10%) at Week 30 in the ITT
population. At week 54, ACR20 was highly similar between groups (CT-P13, 57.0% [172/302]; INX, 52.0% [158/304];
95% CI: -0.03*0.13). The proportion of patients with positive ADAs was comparable between CT-P13 (40.2%, 52.3%)
and INX (39.9%, 49.5%) at week 30 and 54.
PLANETAS and PLANETRA trial reported that TEAEs, ADA and tuberculosis showed similar profile between CT-P13
and INX up to week 54.
Conclusions: CT-P13 demonstrated equivalent pharmacokinetics and efficacy to INX up to week 54. CT-P13 was
well-tolerated, with a safety profile comparable to that of INX up to week 54.
099 RISK MANAGEMENT PLAN IN BIOLOGIC ERA: TAIWAN EXPERIENCE
H.Y. Lin1, W.J. Su2, Y.H. Huang3
1Rheumatology
and Immunology, Taipei Veterans General Hospital, Taipei, Taiwan
Medicine, Taipei Veterans General Hospital, Taipei, Taiwan
3Gastroenterology, Taipei Veterans General Hospital, Taipei, Taiwan
Biologic agents used to treat rheumatic diseases have made a significant impact on the increased risk of serious
infections, especially reactivation of latent tuberculosis infection (LTBI) and hepatitis B virus (HBV).
2Chest
In Taiwan, according to a multicenter, prospective, and observational study, the incidence of LTBI in RA patients is
about 11.6% based on IGRA and 18.6% by TST assay, and a high rate (37.0%) of TST conversion was observed
among patients who had completed 12-month adalimumab therapy. Therefore, patients taking TNFα blockers should
be aware that they are more susceptible to serious TB infections.
HBV reactivation with increase in serum HBV DNA and ALT is an important issue for HBV populations. Reverse
seroconversion from hepatitis B surface antigen (HBsAg)-negative to HBsAg-positive would also happen in hepatitis
B core antibody (anti-HBc)-positive patients. The incidence of HBV reactivation could be as high as 40-60% in HBV
carriers during anti-TNF treatment based on some retrospective studies.
Department of Health and FDA in Taiwan propose the post-marketing risk management plan (RMP) in April, 2012.
Following the policy, Taiwan Rheumatology Association provides a recommendation for LTBI screening and HBV
evaluation. (1) Rheumatic patients who are candidates for biologics should receive LTBI and TB screening; (2) If
patients have LTBI based on the exposure history, CXR findings, or the results of IGRA, may consider of prophylactic
therapy; (3) Routine HBV markers screening, including HBsAg and anti-HBc, is mandatory prior to receiving biologic
agents for all rheumatologic patients.
100 RENOPROTECTIVE EFFECTS OF CITRAL BY MODULATING T CELL FUNCTION, NLRP3 AND
ENHANCING NRF2 ACTIVATION IN AN ACCELERATED LUPUS NEPHRITIS MODEL
S.M. Ka1, S.M. Yang2, K.F. Hua3, J.M. Chang4, L.K. Chao5, C.L. Ho6, A. Chen2
1Graduate
Institute of Aerospace and Undersea Medicine, National Defense Medical Center Taipei, Taipei, Taiwan
of Pathology, Tri-Service General Hospital National Defense Medical Center, Taipei, Taiwan
3Department of Biotechnology and Animal Science, National Ilan University, Ilan, Taiwan
4Department of Animal Pharmacology, Development Center for Biotechnology, Taipei, Taiwan
5Department of Cosmeceutics, China Medical University, Taichung, Taiwan
6Division of Wood Cellulose, Taiwan Forestry Research Institute, Taipei, Taiwan
Lupus nephritis is a leading cause for the morbidity and mortality of systemic lupus erythematosus (SLE). Impaired T
cell function, oxidative stress and renal mononuclear leukocyte infiltration are associated with the acceleration and
deterioration of lupus nephritis, although the exact pathogenic pathway remains largely unclear. Establishment of a
new pathogenesis-oriented therapeutic strategy for the disease is clinically warranted. Citral (3,7-dimethyl-2,6octadienal), a major active compound in a Chinese herbal medicine of Litsea cubeba, can inhibit oxidant activity,
macrophage and NF-κB activation. In the present study, we tested the renoprotective effects of Citral in a mouse
accelerated and severe lupus nephritis (ASLN) model induced by the administration of lipopolysaccharide to SLEprone NZB/W F1 mice. The results show that Citral at a daily dose of 200 mg/kg of body weight by gavage for
consecutive one month significantly ameliorated hematuria, proteinuria, renal function disturbance and severe renal
lesions including neutrophil infiltration and crescent in the glomerulus, and T cell/macrophage infiltration in renal
interstitium. Further mechanistic analyses revealed that Citral negatively regulated T cell function, inhibited the
activation of NLRP3 inflammasome and augmented Nrf2 anti-oxidative activities in the kidney. Our data support that
Citral is renoprotective in the mouse model of severe lupus nephritis by its anti-inflammatory and anti-oxidative
activities.
2Department
Key Words: Systemic lupus erythematosus; lupus nephritis; Litsea cubeba; Citral; neutrophil; NLRP3 inflammasome;
Nrf2
101 FUNCTIONAL ABNORMALITIES AND REGULATORY MECHANISMS OF PLASMACYTOID DENDRITIC
CELLS IN THE DEVELOPMENT OF SYSTEMIC LUPUS ERYTHEMATOSUS
S. Yan1, R.C.Y. Tam1, V.S.F. Chan1, C.S. Lau1
1Medicine,
The University of Hong Kong, Hong Kong, Hong Kong China
Background and aims: Systemic lupus erythematosus (SLE) is a chronic multi-organ autoimmune disease with
immunological features of a prominent Òinterferon (IFN) signatureÓ, which is marked by the elevated expression of
type I IFN-regulated genes in blood and tissue cells of patients with this condition. Plasmacytoid dendritic cells
(pDCs), the most potent type I IFN-producing cells, are previously found to be hyperactive in SLE. Using the New
Zealand Black/White F1 lupus mouse model, the current study sought for the regulatory mechanism of IFN
production by pDCs in SLE.
Methods: Mice with lupus symptoms (symptomatic) such as high titers of serum anti-nuclear antibodies and
persistent proteinuria were compared with the pre-symptomatic ones. Age- and sex-matched non-lupus maternal
NZW mice were used as controls.
Results: While the development of pDCs appeared to be unaffected by lupus, elevated upregulation of MHC class II
and co-stimulatory molecules, and induction of IFN-stimulated gene Ifitm3 in TLR7-stimulated lupus pDCs suggested
phenotypic and functional hypersensitivity of these cells. Furthermore, analysis of the expression profile of
microRNAs in pDCs upon TLR7 activation identified six differentially regulated targets. Among these, miR-155 was
the most highly induced and its induction was consistently higher in pDCs from symptomatic NZB/W F1 mice.
Conclusions: Current investigations pursue on the correlation between up-regulated miRNA-155 induction and
aberrant pDC functions in SLE using miRNA mimics and inhibitors. It is hoped that findings from this study contribute
to a better understanding of SLE pathogenesis and ignite future interests in evaluating the molecular layer of
regulation in autoimmunity.
102 KILLER T CELL INHIBITION BY CD226 BLOCKADE FOR TREATMENT OF AUTOIMMUNE POLYMYOSITIS
N. Tateishi1, S. Hirata1, K. Shibuya2, A. Shibuya2, N. Miyasaka1, H. Kohsaka1
1Medicine
and Rheumatology, Tokyo Medical and Dental University, Tokyo, Japan
of Immunology Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba,
2Department
Japan
(Backgrounds)
Current treatment strategy of polymyositis/dermatomyositis is based largely on successful experiences in treatment of
other systemic autoimmune disorders. To develop a new treatment that addresses specific pathology, CD226
(DNAM-1), which expressed on T cells to promote killer cell function upon interaction with its ligands, was
investigated as a candidate therapeutic target.
(Methods and Results)
Immunostaining of CD226 ligands (CD112 and CD155) disclosed that C2C12 myoblasts and C2C12-derived
myotubes expressed CD155 but not CD112 at the protein levels. In vitro treatment of splenic CD8 T cells with
immobilized monoclonal anti-CD226 antibodies (Tx42) augmented IFN gamma secretion when the T cells were
stimulated with immobilized anti-CD3 antibodies. The augmentation was more obvious when the CD3 antibody
concentration was low. C-protein induced myositis (CIM) we developed as a new polymyositis model was ameliorated
when immunized mice were treated with F(ab')2 of Tx42 both in preventive and therapeutic protocols. Intact Tx42
was not effective in either protocol.
(Conclusions and Discussions)
Muscles express ligands of CD226. Blockade of CD226 with F(ab')2 antibodies, but not with intact antibodies,
suppressed CIM. In vitro agonistic effects of the CD226 antibodies in accelerating CD8 T cell activation suggested
that abrogation of Fc mediated cross-linking by digestion to produce F(ab')2 is essential for the therapeutic effects of
the antibodies. Killer cell inhibition by CD226 blockade should be a new therapeutic approach that addresses specific
pathology of autoimmune myositis.
103 MULTIPOTENT STROMAL CELLS-CLINICAL APPLICATIONS IN AUTOIMMUNE DISEASES AND BEYOND SHOULD AUTISM SPECTRUM DISORDERS BE NEXT?
B. Gesundheit1, D. Naor2, M. Melamed2, J.P. Rosenzweig3
1Autism
Research, Alon Shvut Darom, Alon Shvut, Israel
Center for General and Tumor Immunology, Hebrew University Hadassah Medical School, Jerusalem,
2Lautenberg
Israel
3Autism Research, Feuerstein, Jerusalem, Israel
New directions in Autism Spectrum Disorders (ASD) research suggest that a subgroup should be discussed in the
context of autoimmunity and, as a result, even in context of potential cell therapy. Immunological aspects of ASD will
be summarized, focusing on the cytokine profile of a subset of ASD, along with the rationale for cellular therapy.
Potential safety concerns will be contrasted with hazards in cell therapy for other diseases. Questions for future
research will be posed, highlighting areas demanding further attention.
In recent years, multipotent stromal/stem cells (MSC) have been the subject of intense research owing to their ability
to differentiate into a variety of cell types. By avoiding allorecognition these cells can be used therapeutically for a
spectrum of conditions without triggering rejection by the immune system. Furthermore, in addition to tissue repair
potential, MSCs can serve as regulator cells attenuating autoimmune diseases, including possibly the autoimmune
subset of ASD. In order to assess which conditions can most benefit from innovative cell therapy, preclinical and
clinical research will be reviewed for multiple indications, focusing on mechanism of action. Many mechanisms have
been suggested for how MSCs modulate the immune system. Three main mechanisms, namely the IDO pathway, the
NO pathway, and the PGE2 pathway will be elucidated. The advantages of MSC over other types of cellular therapy
will be stressed along with an investigation of the benefits of allogenic versus autologous cells. The unique
considerations of each indication will be considered, especially in context of ASD.
Parallel Session: Parallel Session 6: Autoantigens and autoantibodies in autoimmune diseases
107 Citrullinated antigens and new therapies for rheumatoid arthritis
R. Thomas1
1Diamantina
Institute, University of Queensland, Brisbane, Australia
Disease modifying strategies are available for treatment of rheumatoid arthritis (RA), and good response rates are
achieved. However, limitations include toxicity, a response rate ceiling, cost and rationing of biologic therapies,
inability to cure or permanently reverse RA pathology, and inability to prevent disease. Recent evidence suggests
that treatment of very early RA with immunomodulatory drugs can delay or attenuate disease onset. RA is strongly
associated with the HLA-DRB1 locus that possesses the Òshared susceptibility epitope (SE)Ó, and the citrullination
of self-antigens. Approximately 80% of RA patients develop autoantibodies targeted against citrullinated self
peptides. These patients are more likely to have RA-associated HLA-DR risk alleles and to smoke. Using HLA-II
tetramers, we demonstrated citrullinated vimentin and aggrecan-specific CD4+ T cells in the peripheral blood of HLADRB1*04:01+ RA patients and healthy individuals, and cytokine responses ex vivo. In RA patients, the number of
autoreactive cells correlated with disease activity. We are developing antigen-specific immunotherapy to target
dendritic cells (DC) in situ with liposomes encapsulating citrullinated peptide and NF-kB inhibitor. In a proof-ofconcept trial, delivery of citullinated peptide and tolerogenic DC was safe and had systemic immune effects. DC
represent an important target for citrullinated peptide-specific immunotherapy in RA.
108 PERFORMANCE OF A FULLY AUTOMATED ASCA TESTING
W. Papisch1, R. Stork2
1International
Sales and Marketing, Phadia GmbH (part of Thermofisher Scientific), Freiburg, Germany
Development, Phadia GmbH (part of Thermofisher Scientific), Freiburg, Germany
Background
ASCA (anti-Saccharomyces cerevisiae antibodies) occur in CrohnÕs disease (CrD), which, besides ulcerative colitis
(UC), is one of the most important inflammatory bowel diseases (IBD). ASCA assist to differentiate between CrD and
UC especially in case of unclear biopsy results. They are produced as IgG and IgA isotypes. ASCA testing is applied
in various ways in the diagnostic procedures.
Objective
To evaluate the performance of a new fully automated ASCA fluoroenzymimmunoassay
Material and Methods
ASCA IgG/IgA were measured in a cohort of 100 CrD, 100 UC and 298 DC patients. Levels of IgG/IgA ASCA were
measured using EliATM ASCA performed on the fully automated Phadia 250 instrument. Results were compared to
findings obtained by two conventional manual Elisa assays. Clinical performance data were calculated .
Results
Clinical performance data obtained by the various assays are outlined in Table 1 .
2Assay
TABLE 1. Best performance outlined in red.
Evaluating the positive likelihood ratios 2/3 assay combinations achieve a clinical usefulness (PLR > 5).
The results demonstrate that the diagnostic usefulness of ASCA testing is highly dependent on the assay used, as
well as on the combination of isotypes (IgG/IgA).
Conclusion




Combination of IgG and IgA is required to achieve clinical usefulness
High specificity of the test used is crucial for valuable diagnostic support
The new fully automated EliATM ASCA tests demonstarte an overall suprior performace compared to
2 Elisa assays.
Carrying out ASCA testing embedded in a reasonable diagnostic workup can assure the diagnosis
and support the diagnostic process for the clinician
109 EVALUATION OF A NEW DOT BLOT FOR THE PARALLEL DETECTION OF 12 ANTINUCLEAR
ANTIBODIES INCLUDING DFS70
J. Schulte-Pelkum1, E. Tena1, Z. Shums1, G. Norman1, M. Mahler1
1R&D,
INOVA Diagnostics Inc., San Diego, USA
Background and aims: The screening for antinuclear antibodies is the initial step for the serological analysis of
patients suspected to suffer from systemic autoimmune rheumatic diseases (SARD). Recently, antibodies against
DFS70 have been described as the most common cause of positive ANA screen results on HEp-2 cells without any
clinical association with SARD. We present the evaluation of a new 12plex Dot blot assay with DFS70 antigen using a
digital reading system.
Methods:
Sera (n=140) from SARD patients were tested both using individual QUANTA Flash® or QUANTA Lite® assays for
the respective autoantibodies (all INOVA Diagnostics) and by the new ANA Dot Blot assay. Interpretation was done
using a digital dot recognition system, consisting of a quick load tray for the dot blot strips and a special scanning
device. Agreements between QUANTA Flash/QUANTA Lite and the Dot Blot assay were calculated for the individual
antigens.
Results:
High agreements of the methods were observed: Positive (PPA), negative (NPA) and total percent agreements (TPA)
were close to or higher than 85%. ROC analysis against established tests resulted in high AUC values for all antigens
(e.g. DFS 70: AUC=0.82, PPA=88.9%, NPA=91.7% and TPA=91.3 %)
Conclusion:
The new ANA Profile assay in combination with the scanning system offers fast and reliable multiplex detection of 12
ANAs including anti-DFS70 antibodies. The system shows good correlation to established immunoassay systems
and allows for the interpretation of often unresolved IIF results in patients without clinical evidence of SARD.
110 THE EFFECT OF CXCL10 BLOCKADE IN C PROTEIN-INDUCED MYOSITIS
J. Kim1, J. Choi2, S.H. Park3, S.H. Yang4, J.A. Park2, K. Shin2, E.Y. Lee2, E.B. Lee2, H. Kawachi5, H. Kohsaka6, Y.W.
Song7
1Internal
Medicine, Chungnam National University Hospital, Daejeon city, Korea
Medicine, Seoul National University College of Medicine, Seoul, Korea
3Pathology, Seoul National University College of Medicine, Seoul, Korea
4Kidney Research Institute, Seoul National University, Seoul, Korea
5Cell biology Institute of Nephrology, Niigata University Graduate School of Medical and Dental Sciences, Niigata,
Japan
6Medicine and Rheumatology, Tokyo Medical and Dental University, Tokyo, Japan
7Internal Medicine, Seoul National Univerisity College of Medicine, Seoul, Korea
Backgrounds and aims: CXCL10 (also called interferon-γ-inducible protein 10 [IP-10]) is a chemokine that plays a
critical role in the infiltration of T cell in autoimmune disease. CXCL10 is reported to be expressed in muscle tissue of
polymyositis. We investigated the role of CXCL10 and the effect of CXCL10 blockade in C protein-induced myositis,
an animal model of polymyositis.
2Internal
Methods: C protein-induced myositis model was induced with human skeletal C protein fragment in 8-week-old
female C57BL/6 mice. Immunohistochemistry was performed to detect CXCL10 and CXCR3, its receptor. CXCR3 in
mouse splenocyte was investigated by flow cytometry and migration assay of mouse splenocyte was performed. Mice
with C protein-induced myositis were treated with anti-CXCL10 antibody or control IgG 8 days after the induction of
myositis every other day and the muscle inflammation was assessed 3 week after the induction.
Results: Immunohistochemistry showed the expression of CXCL10 and CXCR3 in the muscle of C protein-induced
myositis. Invading CD8+ T cell into myofiber expressed CXCR3. Moreover,IFN-γ+ cells were increased among
CXCR3+CD8+ T cells compared to CXCR3-CD8+ T cells (CXCR3+CD8+ T cell, 28.0 ± 4.2% vs. CXCR3-CD8+ T
cell, 9.5 ± 1.5%, p = 0.016). Migration of splenocyte was increased in response to CXCL10 (chemotactic
index=1.91±0.45). Treatment with anti-CXCL10 antibody (n=10) showed less inflammation score in muscles than
treatment with control IgG (n=10; median [range], anti IP-10, 0.75 [0.25-2.00] vs. control IgG, 1.43 [1.125-4.25],
p=0.045).
Conclusion: CXCL10 was expressed in the inflammation of C protein-induced myositis model and its blockade
suppressed inflammation in muscle.
111 ANTI-MYELIN OLIGODENDROCYTE GLYCOPROTEIN ANTIBODY IN PAEDIATRIC CNS DEMYELINATION
DEFINES AN AUTOIMMUNE TREATMENT-RESPONSIVE INFLAMMATORY CNS DEMYELINATION
REMINISCENT OF NMO SPECTRUM DISORDER.
R.C. Dale1, V. Merheb1, E. Tantsis1, R.Y.A. Kumaran1, F. Brilot1
1Institute
for Neuroscience and Muscle Research, The Children's Hospital at Westmead, Westmead, Australia
Background and aims. Over the few last years, anti-human surface myelin oligodendrocyte glycoprotein (MOG)
immunoglobulin G antibody (MOG antibody) have been detected in approximately 20-40% of paediatric patients with
first episode of CNS demyelination, but also in a few patients with Neuromyelitis Optica (NMO) spectrum disorder
who are negative for NMO antibody. However, although the existence of this MOG antibody-positive paediatric
subgroup is largely accepted and has been shown by multiple teams, the utility and the relevance of these antibodies
are still unknown. We aimed to determine the importance of MOG antibody in paediatric patients with inflammatory
CNS demyelination and whether MOG antibody could have pathogenic effect on oligodendrocyte physiology.
Methods. We have performed detailed clinical phenotyping in a large Australian cohort of 75 retrospective and
prospective children with CNS demyelination. We have measured MOG antibody using a flow cytometry lentivirustransduced MOG-expressing cell line. Positivity for MOG antibody was defined as three standard deviations above
the mean control value. We have compared antibody positive and negative patients. We also have tested the in vitro
pathogenic potential of these antibodies using human oligodendrocytes.
Results. We have found 40% of children positive for MOG antibody (30/75) and 0% in controls (0/22). MOG
antibody-positive patients were more likely to be young, have acute disseminated encephalomyelitis, have bilateral
optic neuritis (but not unilateral), have longitudinal extensive transverse myelitis, and have an elevated erythrocyte
sedimentation rate. There was a strong correlation between MOG antibody positivity and bilateral optic neuritis, as
well as between MOG antibody and erythrocyte sedimentation rate. In relapsing patients, intrathecal oligoclonal
bands were only seen in MOG antibody-negative patients. Recent prospective relapsing MOG antibody- positive
patients have been treated with steroids and mycophenolate. These patients were responsive to immune therapies,
and levels of MOG antibody decreased with treatment. When we assessed the pathogenic potential of MOG antibody
on live human oligodendrocytes, we observed a reduced MOG surface staining in otherwise healthy-looking cells,
whereas live cells treated with control antibody displayed an undisturbed surface MOG expression.
Conclusions. Overall our data suggest that there is emerging evidence that MOG antibody defines a separate
autoimmune inflammatory disorder with a phenotype more reminiscent of NMO spectrum disorders and that MOG
antibody might be pathogenic in these patients.
112 A NON-INVASIVE PET/CT IMAGING PROCEDURE FOR DIAGNOSING ATHEROSCLEROTIC LESIONS:
IMMUNOLOGIC TARGETING TO OXLDL/BETA2-GLYCOPROTEIN I IN THE WHHL RABBITS
T. Sasaki1, F. Takenaka1, Y. Matsunami2, H. Hirano3, S. Kita2, M. Eiji4
1Collaborative
Research Center for Okayama Medical Innovation Center (OMIC), Okayama University Graduate
School of Medicine Dentistry and Pharmaceutical Sciences, Okayama, Japan
2IVD Development Department, Medical and Biological Laboratories Co Ltd, Ina, Japan
3Engineering Department, SHI Accelerator Service Ltd, Tokyo, Japan
4Department of Cell Chemistry, Okayama University Graduate School of Medicine Dentistry and Pharmaceutical
Sciences, Okayama, Japan
Background: Current in vivo diagnostic tools for detecting vulnerable plaques in preclinical and/or autoimmune
atherosclerotic patients required a range of non-invasive imaging systems. We have demonstrated that oxidized LDL
(oxLDL) form complexes with β2-glycoprotein I (oxLDL/β2GPI complexes) that function as a major proatherosclerotic/pro-thrombogenic autoantigen capable of triggering antiphospholipid antibodies. The aim of the study
was to confirm the clinical utility of 64Cu-labeled single chain Fv of anti-oxLDL/β2GPI antibody (3H3-scFv) as a PET
imaging probe for atherosclerotic lesions in the Watanabe heritable hyperlipidemic (WHHL) rabbits. Methods and
Results: Twenty-four hours prior to obtaining PET/CT fusion images, 3H3-scFv labeled with 64Cu was intravenously
injected into WHHL and control rabbits. The PET/CT fusion images demonstrated that aortic arches were clearly
visualized only in WHHL but not control rabbits. Aortic arches and thoracic/abdominal arteries were positively stained
with Sudan IV in WHHL but not control rabbits. After obtaining the PET/CT images, autoradiographic analyses and
bio-distributional determination of 3H3-scFv were performed. Those results clearly revealed that 3H3-scFv was
specific and preferentially accumulated in atherosclerotic plaques to provide an accurate imaging detection of the
lesion. The bio-distribution analyses also quantitatively showed that the accumulation of 3H3-scFv was 1.37 to 2.81fold greater in aortas of WHHL than control rabbits. Conclusions: Because of the similarity between the size of rabbit
aortas and human coronary arteries, 3H3-scFv (an antibody variant lacking the Fc region) can be used as a probe
suitable for in vivo clinical PET imaging of atherosclerosis with highly enough sensitivity and specificity.
113 FILAMENTOUS ANTIGEN-SPECIFIC THERAPY OF EAE VIA INTRANASAL DELIVERY OF PHAGE
DISPLAYING A MYELIN IMMUNODOMINANT EPITOPE
B. Solomon1
1Microbiology-Biotechnology,
Tel-Aviv university, Tel-Aviv, Israel
Backgound and aims
The presence of anti-myelin antibodies in patients with early multiple sclerosis (MS) and in MS animal models led to
renewed interest in a role for B cells, plasma cells and their products in the pathogenesis of the disease. Here we
propose a novel strategy based on engineered filamentous phage in which its major coat protein was fused to the
immunodominant epitope derived from the myelin oligodendrocyte glycoprotein (MOG 37-44). Filamentous phages
are well-studied, both structurally and genetically. Their shape as a long fiber, 1000nm long and 6nm wide, enables
penetration to the central nervous system via nasal administration.
Methods: Experimental autoimmune encephalomyelitis (EAE) diseased mice (as a model of MS) intranasally treated
with phage-MOG .
Results
The treated animal models showed improved clinical scores; reduction of antibodies against MOG; reduced
proinflammatory cytokines, in particular monocyte chemoattractant protein 1 (MCP-1), interferon g (IFN-g) and IL-6;
and prevented demyelinization, compared to untreated animals.
Conclusions
Brain delivery of MOG via filamentous phages suggests that the improved clinical effects obtained in EAE mice may
be due to depletion of MOG autoantibodies in situ and/or stimulation of immune mechanisms towards induced
tolerance in the periphery, indicating that the humoral immune system in MS would be a reasonable therapeutic
option.
Plenary Session: Plenary Session 3: Genetics and Immunogenetics of autoimmune diseases
118 The pebbles of autoimmunity: genetic and autoantibodies in the complex mosaic of autoimmunity
C. Perricone1, N. Agmon-Levin2, F. Ceccarelli1, G. Valesini1, J.M. Anaya3, Y. Shoenfeld2
1Dipartimento
di Medicina Interna e Specialità Mediche, Sapienza Università di Roma, Roma, Italy
Faculty of Medicine Tel-Aviv University, The Zabludowicz Center for Autoimmune Diseases Sheba Medical
Center, Tel-Aviv, Israel
3Center for Autoimmune Diseases Research (CREA), Universidad del Rosario, Bogotà, Colombia
Autoimmune diseases (ADs) are chronic conditions initiated by the loss of immunological tolerance to self-antigens.
The pathogenic hypothesis comprises a complex interaction between genetic, environmental and hormonal factors
that interact in an individual over time generating a dysregulation of the immune system leading to disease
development. Several polymorphic genes contribute to the development of ADs. Furthermore, age and gender play a
major role by influencing hormone levels that can represent the fulcrum unbalancing from susceptibility to protection.
Evidences suggest that while all these steps occur, the susceptible individual develops autoantibodies over a long
time lapse. Such autoantibody production is genetically determined and finally, their presence seems to determine
the clinical presentation of ADs. The genetic predisposition to the developments of autoantibodies and toward the
disease process may overlap.
2Sackler
119 Antiphospholipid syndrome; 30 years
In 1983, Graham Hughes first described the concept of antiphospholipid syndrome (APS). The term APS refers to
patients with arterial and/or venous thrombosis, pregnancy complications and circulating antibodies called
Òantiphospholipid antibodiesÓ, appearing to make the blood far more sticky and liable to clot. In 1984, we described
the enzyme-linked immunosorbent assay (ELISA) system which directly detected circulating aCL in patients with SLE
who revealed biological false positive STS for syphilis. In 1990, three groups independently reported the necessity of
a cofactor for the binding of autoimmune anticardiolipin antibodies (aCL) to the solid phase phospholipids. β2glycoprotein I (β2GPI) was identified as this cofactor. In 1994,the epitope for aCL was shown to develop when β2GPI
is adsorbed on polyoxygenated polystyrene plates. In 2000, we described antiprothrombin antibodies bind to
prothrombin exposed to immobilized phosphatidylserine and established a phosphatidylserine dependent monoclonal
antiprothrombin antibody. In 2004, a novel role of nicked β2GPI was identified in the negative feedback pathway of
extrinsic fibrinolysis. Nicked β2GPI was found to bind angiostatin 4.5 and to attenuate its antiangiogenic property. In
2004, it was shown that the p38 MAPK pathway mediates induction of the tissue factor (TF) gene in stimulated with
human monoclonal anti- β2GPI antibodies. Very recently, β2GPI was identified as a complement regulator. The
cross-link between complement activation and prothrombotic status in patients with APS will be discussed.
120 From genetics to functional insights into the pathogenesis of rheumatoid arthritis
K. Yamamoto1, A. Suzuki2, Y. Kochi2
1Department
of Allergy and Rheumatology, Graduate School of Medicine The University of Tokyo, Tokyo, Japan
for Autoimmune Diseases, Center for Integrative Medical Sciences RIKEN, Yokohama, Japan
The etiology and pathogenesis of rheumatoid arthritis (RA) are still unknown. Genome-wide association study
(GWAS) has been conducted to identify RA associated genes. However, genetic study does not necessarily provide
a direct conclusion. Therefore, information from GWAS should be used to understand the functional mechanisms of
RA. For example, HLA-DR4 has been reported as the strongest genetic factor in RA. However, the precise
mechanism of DR4 in RA pathogenesis has been unknown.
2Laboratory
We reported functional haplotypes of PADI4, encoding citrullinating enzyme peptidylargimine deiminase 4, are
associated with RA. Our results imply that the RA susceptible PADI4 haplotype increases production of citrullinated
peptides acting as auto-antigens. The function of PADI4 in inflammatory arthritis was further studied using knock-out
mice. The knock-out mice exhibited decreased inflammatory joint lesions in collagen-induced arthritis, suggesting that
PADI4 is actually involved in the pathogenesis of RA.
We also reported a polymorphism that up-regulates the expression of Fc receptor-like 3 (FCRL3) gene. This
polymorphism has recently been confirmed as predisposing for several different autoimmune diseases. FCRL3 is
preferentially expressed on B cells. We found FCRL3 potentially inhibits BCR-mediated signaling. These results
suggest that augmented inhibition of BCR-mediated signaling by FCRL3 alter the activation threshold and promote
tolerance breakdown in B cells.
We have further identified a polymorphism in the chemokine receptor CCR6 gene is associated with RA. CCR6 is a
surface marker for Th17 cells, suggesting that CCR6 is critically involved in Th17-driven autoimmunity in RA.
121 Aberrant Chromatin Modifications: The Epigenetics Mechanisms of Lupus
Epigenetic factors are important in the onset of human disease such as cancer, aging and autoimmunity. We aimed
to investigate the aberrant epigenetic regulation in lupus, and found that T cells from patients with active lupus have
genome-wide decrease in deoxymethylcytosin, and gene-specific hypomethylation. DNA hypomethylation induces
overexpression of CD11a (ITGAL), perforin (PRF1), CD70 (TNFSF7) and CD40LG (TNFSF5) genes, and causes
autoreactive monocyte/macrophage killing and excessive B cell stimulation. We also found global histone H3 and H4
hypoacetylation and H3K9 hypomethylation in active lupus CD4+ T cells compared with controls. We provide
evidence indicating that the transcription factor RFX1 down-regulation appears to be critical event leading to DNA
methylation and histone acetylation defects in T cells of patients with lupus. In addition, we demonstrated eleven
miRNAs to be significantly increased or decreased in SLE CD4 + T cells relative to controls. Among these, miR-126
was upregulated and the degree of overexpression was inversely correlated with DNMT1 protein levels. We
demonstrated that overexpressing miR-126 in healthy donor CD4+ T cells caused the demethylation and upregulation
of genes encoding CD11a and CD70, thereby causing T and B cell hyperactivity. Inhibiting miR-126 in SLE CD4+ T
cells had opposite effects.
In summary, aberrant chromatin modification plays a critical role in the development of lupus, and epigenetic-based
therapies appear to hold promise for the treatment of lupus.
Parallel Session: Parallel Session 7: Challenges in SLE
129 IMMUNOPATHOLOGICAL ROLES OF PATTERN-RECOGNITION RECEPTORS IN SYSTEMIC LUPUS
ERYTHEMATOSUS
C.K. Wong1, L.S. Tam2, S.L. Yu2, P.T.Y. Wong1, D.P. Chen1, E.K. Li2
1Department
of Chemical Pathology, The Chinese University of Hong Kong, Shatin, Hong Kong China
of Medicine and Therapeutics, The Chinese University of Hong Kong, Shatin, Hong Kong China
Background: Pattern recognition receptors (PRRs) including Toll-like receptors (TLR), NOD-like receptors (NLRs) and
receptor for advanced glycation end products (RAGE) play import roles for the innate immunity against exogenous
pathogens and endogenous damages. Methods: We investigated the expressions and functions of PRRs of PBMC in
SLE patients using flow cytometry and immunoassays. Results: The aberrant expression of TLR and TLR-mediated
release of inflammatory cytokines/chemokines was observed in SLE patients. Over-expression of intracytosolic
NOD2 in monocytes of na•ve SLE patients treated with immunosuppressant exhibited positive association with longer
disease duration. Immunosuppressive therapies could down-regulate the NOD2 expression in CD8+T lymphocytes,
monocytes, myeloid and plasmacytoid dendritic cells in SLE patients which subsequently reduce IL-10 production,
contributing towards the immunopathological mechanisms of SLE. Expression of full-length RAGE (flRAGE) was
significantly increased in the monocytes of SLE patients, correlating with the plasma RAGE ligand high mobility group
box (HMGB)1 levels. Plasma soluble (s)RAGE level negatively correlated with SLE disease activity. Plasma
endogenous secretory RAGE (esRAGE) level was significantly lower in SLE patients with flare while esRAGE/sRAGE
ratio negatively correlated with complement level. HMGB1 could moderately induce ex vivo inflammatory IL-6
production, resulting in transient activation of intracellular p38 MAPK, JNK and NF-κB in peripheral blood monocytes.
Moreover, TLR9 ligand together with HMGB1 exhibited a synergistic effect on IL-6 and IL-12 secretions. Conclusion:
The immunopathogenesis of SLE is related to the aberrant expression and function of PRRs and plasma sRAGE may
serve as a potential biomarker for disease activity and future therapeutic target in SLE.
2Department
131 PLASMACYTOID DENDRITIC CELLS IN SLE PATHOGENESIS
V. Chan1, S. Yan1, C.S. Lau1
1Medicine,
University of Hong Kong, Hong Kong, Hong Kong China
Plasmacytoid dendritic cells (pDCs) constitute a unique subset of cells which acquire many features of conventional
dendritic cells (cDCs) upon activation. Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease
affecting predominantly young females associated with significant morbidity and mortality. While cDCs likely
contribute to SLE pathogenesis by breaking immune tolerance to self-antigens, pDCs are thought to play a major
pathogenic role by virtue of its unique ability to produce large amount of type-I interferon upon nucleic acids
stimulation through the toll-like receptors (TLRs)-7/9. We have previously observed a significant increase in the
frequency of pDCs which show enhanced T cell stimulatory activity in SLE patients. Remarkably, bone marrowderived pDCs from patients also exhibit activated phenotypes with higher expression of co-stimulatory molecules and
induce a stronger T-cell proliferation. In the NZB/W F1 mouse model, we have similarly observed that bone
marrow*derived pDCs from symptomatic F1 mice exhibit heightened up-regulation of co-stimulatory molecules upon
TLR-7 stimulation when compared with pre-symptomatic animals. Functional abnormalities have also been found. In
the study of regulatory mechanisms of pDC abnormality in lupus, microRNAs (miRNAs) expression analysis reveals
that distinct miRNAs are differentially expressed by pDCs from lupus mice. On-going experiments are underway to
correlate the perturbed miRNAs induction and the aberrant pDC functions in SLE using miRNA mimics and inhibitors.
In summary, pDC functional abnormalities clearly contribute to SLE development and the study of regulatory miRNAs
in lupus pDCs may reveal new potential therapeutic targets for clinical intervention.
133 THE ALARMIN FUNCTION OF IL-33/ST2 AXIS CONTRIBUTES TO ENDOTHELIAL CELL INJURYMEDIATED PROINFLAMMATORY RESPONSE IN LUPUS NEPHRITIS
T. Yi1, Y. Shui-Lian1, H. Can-Hui1, H. Wen-Hui1, W. Zhuo-Long1
1Department
of Rheumatology, Guangzhou Medical University, Guangdong, China
Background: "Alarmins" are prototypic endogenous pro-inflammatory factors are released from necotic cells and
provoke local damage or systemic inflammation. Evidences are accumulating to support the inclusion of "Alarmins"
as targets of autoreactivity as well as inducers in the pathogenesis of Systemic Lupus Erythematosus (SLE).
Interleukin (IL)-33 is a novel member of the family of "Alarmins" because of its characteristics and functions in
mediating host immune responses. We sought to determine the role of IL-33/ST2 axis in lupus pathogenesis.
Objectives: (1) To determine whether IL-33 was present in renal glomerular endothelial cells; (2) To assess the
functional and intracellular signal transuction mechanisms regulating the link between IL-33/ST2-mediated innate
immunity and inflammation in CD4+ T cells-endothelial cells co-culture system of lupus patients.
Preliminary results: Immunofluorescence (IF) for IL-33 in the kidney were performed in both MRLlpr lupus mice and
C57BL/6J mice. On double staining for IL-33 and lectin, IL-33 was clearly seen in glomeruli and also in peritubular
areas. To determine whether the IL-33 staining in glomeruli and peritubular areas was in endothelium, double IF
staining for IL-33 and von Willebrand factor (vWF) was performed. IL-33 co-localized with vWF in glomeruli and in
peritubular areas. The increased levels of IL-33 mRNA transcripts were detected in kidney of MRLlpr lupus mice
compared with C57BL/6J mice. Expression of cell-surface ST2 was increased on the CD4+ T cells of lupus patients
when compared with healthy controls. Serum sST2 level was significantly higher in SLE patients with flare than those
without flare
134 CXCL16 EXPRESSION IN SYSTEMIC LUPUS ERYTHEMATOSUS AND THE INFLUENCING FACTORS
ABOUT LUPUS ACTIVITY
T. ZhiMing1, L. ZhuoFeng2, Z. Yu1, L. ZhiFen1, C. YuHang1, W. ZhiWen1, Z. ShiYong1
1Department
of rheumatology, The central people’s hospital of HuiZhou City, HuiZhou, China
of Pharmacy, Wenzhou Medical College, Wenzhou, China
Objective: To investigate the relationship between the change of serum CXC chemokine ligand 16(CXCL16)with
lupus activity in SLE patients and lupus mice.
2Department
Methods: Through clinical observation and animal experiment, 20 SLE patients,20 normal subjects were observed.
Height, weight, waist circumference, age, gender, blood glucose, C-peptide , levels of CXCL16 , ceatinine and Creactive protein(CRP) were obtained.
In Fas-/-lupus mice,CXCL16 -/-C57BL/6J mice?normol C57BL/6J mice (20 in each group), We Compare with the
control group and model group, observate them 4 week, collecte and determine including serum CXCL16, levels of
antibodies to double-strand DNA, oxidized phospholipids, C-reactive protein, as well as IL-6 and TNF-α. To analyze
the clinical datas, studentÕs unpaired t-test was used for the comparison between 2 groups. One-way ANOVA assay
and multiple stepwise regression were used for multiple groups.
Results: Serum level of CXCL16 is significantly increased in SLE patients compared with the healthy control subjects
(P<0.05). Serum levels of CXCL16 of Fas-/-lupus mice are higher than the control group(P<0.05). Serum levels of
CXCL16 of model group are higher than the control group(P<0.05). Multiple stepwise regression analysis indicated
that levels of CXCL16 are positive correlation with lupus activity index such as CRP and antibodies to ds-DNA.
Conclusions: Both of clinical observation and animal experiment indicate Serum CXCL16 levels are positive
associated with activity levels of SLE. But how to elucidate the pathophysiologcial mechanism of CXCL16 in SLE
requires further research.
135 DETERMINING THE FUNDAMENTAL ROLE OF TLR7 IN SYSTEMIC LUPUS ERYTHEMATOSUS
T. Celhar1, J. Dayalan1, R. Magalhaes1, H.Y. Lee1, S.H. Hwang2, J.E. Connolly1, E.K. Wakeland2, A.M. Fairhurst1
1Immunology,
Singapore Immunology Network (SIgN), Singapore, Singapore
UT Southwestern Medical Center, Dallas, USA
Systemic Lupus Erythematosus is a chronic autoimmune disease with complex etiology. It is characterized by the
presence of anti-nuclear autoantibodies (ANA). These form self-reactive immune complexes which deposit in tissues
where they initiate inflammation and the destruction of organs, such as the kidney. Recent data from multiple murine
models has demonstrated that Toll-like receptor (TLR)-7, is involved in both the breakdown in central tolerance and
the progression to severe disease. Previous work from multiple groups has demonstrated that the Yaa murine lupus
susceptibility locus drives the development of glomerulonephritis (GN) via an increase in expression of TLR7.
We have recently described a novel conditional low copy TLR7 transgenic model system (Tg7). These mice carry 2
additional copies of TLR7 and when present on a Sle1 autoimmune background (B6.Sle1Tg7) develop severe
disease (Hwang et al., J Immunol, 2012). Normalization of TLR7 expression solely in B cells restricts the ANA profile
and reduces severe severity. We have now examined the requirement of DC-dependent TLR7 expression for the
development of autoimmunity. We have used the CD11c-Cre recombinase system to normalize the TLR7 expression
solely within dendritic cells (DCs). We demonstrate that the elimination of the additional TLR7 copy in DCs prevents
the development of many autoimmune traits. In addition, we have characterized the infiltrating DCs in the kidney
which are likely to interact with autoantigens, such as components of neutrophil extracellular traps, which are known
to contain TLR7 ligands.
2Immunology,
Parallel Session: Parallel Session 8: Regulation and immunomodulation of autoimmune diseases
139 Vitamin D and systemic lupus erythematosus: an update
C.C. Mok1
1Medicine,
Tuen Mun Hospital, Hong Kong, Afghanistan
Vitamin D is a steroid hormone that exhibits a number of regulatory effects on growth, proliferation, apoptosis and
functions of various cell types of the human immune system on top of its well-known actions on calcium homeostasis
and bone metabolism. The immunomodulatory actions of vitamin D on the immune system are highly relevant to the
pathophysiology of systemic lupus erythematosus (SLE). Low vitamin D state is common in patients with SLE
because of a number of actors that include avoidance of sunshine, sun protection, renal dysfunction and the use of
medications such as glucocorticoids, anti-convulsants. anti-malarials and the calcineurin inhibitors, which either alter
the metabolism of vitamin D or down-regulate the functions of the vitamin D receptors. Hypovitaminosis D in SLE
correlates with disease activity scores, and is associated with low bone mineral density, fatigue and increased
cardiovascular risk. This talk updates the recent evidence on the interplay between vitamin D status and SLE
regarding disease onset, activity state and disease-related complications. Recommendations for vitamin D
supplementation in SLE patients will also be discussed.
140 Novel regulatory T cells controlling autoantibodies
K. Yamamoto1, T. Okamura1, S. Sumitomo1, K. Morita1, Y. Iwasaki1, K. Fujio1
1Department
of Allergy and Rheumatology, Graduate School of Medicine The University of Tokyo, Tokyo, Japan
Dysregulated activation of autoreactive B cells is critical for the development of autoimmune diseases. Autoantibodies
produced by such B cells can induce a variety of autoimmune diseases, including HashimotoÕs thyroiditis, systemic
lupus erythematosus (SLE) and others. The physiological mechanisms responsible for the maintenance of B cell
tolerance by T cells with regulatory activity remain to be investigated. Recently, we identified early growth response
gene 2 (Egr2)-controlled CD4+CD25-Foxp3- regulatory T cells (Treg) that express lymphocyte activation gene 3
(LAG3). LAG3+ Treg effectively suppress autoantibody production and autoimmunity. Adoptive transfer of LAG3 +
Treg, but not CD4+CD25+ Foxp3+ Treg, significantly suppressed the progression of nephritis and autoantibody
production in MRL/lpr lupus-prone mice. LAG3+ Treg strongly suppressed antibody production and the development
of germinal center B cells in a programmed death 1 (PD-1)- and Egr2-dependent manner. Interestingly, Fas/FasL costimulation was required for the full-expression of Egr2 in LAG3+ Treg. These findings indicate an unexpected role of
Fas/FasL interaction in the pathogeneses of autoimmune diseases. From these results, we believe that a novel
regulatory T cell subset (LAG3+ Treg) is one of essential elements in the regulation of the immune responses
involved in the maintenance of self-tolerance.
142 Regulatory B cells in autoimmune arthritis
L. Lu1
1Pathology,
The University of Hong Kong, Hong Kong, Hong Kong China
Regulatory B cells in autoimmune arthritis
Liwei Lu, Department of Pathology, The University of Hong Kong, Hong Kong
Abstract
TNF cytokines play a crucial role in immune regulation by modulating lymphocyte proliferation and apoptosis. Our
previous studies have shown that B cell activating factor (BAFF), a member of TNF family cytokines produced by
myeloid lineage cells such as macrophages and dendritic cells, was critically involved in promoting B cell survival and
autoantibody production in mice with collagen-induced arthritis (CIA). Recently, we identified a novel function of BAFF
in the induction of IL-10-producing regulatory B cells. BAFF-induced IL-10-producing B cells showed a distinct
CD1dhiCD5+ phenotype mainly derived from marginal-zone B cells, which possessed a potent function in inhibiting T
cell activation and cytokine production. BAFF was found to activate the transcription factor AP-1 for binding to IL-10
promoter. In collagen-immunized mice, adoptive transfer of BAFF-induced IL-10-producing B cells markedly reduced
the disease severity and joint damage of autoimmune arthritis via suppression of Th17 cell response. Taken together,
our findings have provided further insight in understanding the roles of BAFF and regulatory B cells in autoimmune
pathogenesis, which may facilitate the development of new therapeutic strategies for targeting autoimmune diseases.
143 THE REGULATION OF DENDRITIC CELL SURVIVAL AND FUNCTION BY TUMOUR NECROSIS FACTOR
RECEPTORS 1 AND 2
N.J. Maney1, A. Krippner-Heidenreich1, C.M.U. Hilkens1
1Musculoskeletal
Research Group, Institute of Cellular Medicine, Newcastle upon Tyne, United Kingdom
Tumour Necrosis Factor (TNF) is a key mediator of inflammatory diseases such as rheumatoid arthritis and plays a
central role in dendritic cell (DC) biology. Our aim was to dissect the individual contributions of the two TNF receptors
(TNFR1 and TNFR2) in regulating the survival and function of human inflammatory monocyte-derived (moDC) and
steady-state myeloid DC. A DNA binding assay was used to demonstrate that in moDC TNFR1, but not TNFR2,
activates the classical p65 NFκB pathway whereas both TNFR1 and TNFR2 activate the alternative p100/p52 NFκB
pathway, highlighting differences in signalling downstream of the receptors. In moDC, TNFR1-selective, but not
TNFR2-selective stimulation resulted in increased expression of DC maturation markers CD83 and CD86, and
enhanced T cell stimulatory capacity. Furthermore, moDC survival was prolonged by selective stimulation of either
TNFR1 or TNFR2 as shown by reduced intracellular levels of active caspase-3, indicating that innate signals can
promote DC survival in the absence of DC maturation. Accordingly, the p65 NFκB pathway was involved in the prosurvival effect of TNFR1 whereas the Bcl-2/Bcl-xL pathway was essential to survival mediated by both TNFR. In
contrast, in myeloid DC, maturation was mainly mediated through TNFR1, whereas TNFR2 was superior in protecting
DC from cell death. An antagonistic TNFR1-specific antibody was used to confirm that cell death protection via
TNFR2 was independent of TNFR1-mediated signalling. Understanding the immunoregulatory properties of signalling
through these two TNF receptors is important for the design of more targeted anti-TNF therapy.
Parallel Session: Parallel Session 9: Autoimmune diseases
146 Liver-resident NK cells in autoimmune skin-contact hepersensitivity
Z. Tian1, H. Peng1, X. Jiang1, Y. Chen1, D. Sojka2, H. Wei1, R. Sun1, W. Yokoyama2
1Institute
of Immunology School of Life Science, University of Science and Technology of China, Hefei, China
Division Department of Internal Medicine, Washington University School of Medicine, St. Louis, USA
Natural killer (NK) cells are multifunctional lymphocytes with divergent phenotypic properties in different organs.
Recent studies has uncovered that liver NK cells possess memory-like properties in contact hypersensitivity (CHS)
models, which challenges the traditional definitions of innate and adptive immunity. However, the phenotype and
origin of these ÒmemoryÓ NK cells cannot be distinguished from other NK cell subpopulations. Here, we defined the
transcriptional, phenotypic and functional features of liver NK cell subsets and their roles in mediating CHS. Liver NK
cells can be divided into two distinct subsets, CD49a +DX5- and CD49a-DX5+. Substantial transcriptional and
phenotypic differences existed between liver CD49a+DX5- NK cells and other NK cell subsets; the former possessed
memory potential and conferred hapten-specific CHS responses upon hapten challenge. Importantly, CD49a +DX5NK cells were liver-resident, present in the liver sinusoid blood but not the afferent and efferent blood of the liver,
whereas CD49a-DX5+ NK cells are highly migratory and incapable of mediating CHS responses. Moreover,
CD49a+DX5- subset appeared to originate from hepatic hematopoietic progenitor/stem cells but not from the bone
marrow, and maintained their phenotypes at steady state. Our findings of liver-resident NK cells shed new light on
understanding how NK cells contribute to skin-contact hepersensitivity.
2Rheumatology
147 HYPOCOMPLEMENTEMIC VASCULITIDES
R. Perricone1, E. Ballanti1
1Department
of Internal Medicine Unit of Rheumatology, University of Rome Tor Vergata, Rome, Italy
The vasculitides are a set of related disorders characterized by blood vessel inflammation leading to tissue or organ
injury. Specific vasculitic disorders are defined depending on the vessels affected (size, type, location). Many
vasculitides are caused by immune complexes and the activation of the complement system can be involved in the
pathogenesis. In particular, complement involvement was demonstrated in small vessel vasculitides, such as ANCAassociated vasculitides, cryoglobulinemia, Henoch-Sch*nlein purpura and IgA nephropathy. Evidences of
complement activation come from the detection of complement components in affected tissues and from the
observation of reduced plasma levels of complement intact proteins. However, the consumption of complement
factors is mainly evident in urticarial vasculitis (UV), a relatively rare clinico-pathologic entity characterized by
recurrent episodes of urticaria. In contrast to other forms of urticaria, the typical wheals of UV persist for more than 24
hours and hyperpigmentation residues after resolution. Among subjects affected, hypocomplementemic patients
show more severe multi-organ involvement than normo-complementemic patients. Anti-C1q autoantibodies have
been described in these patients. Circulating antigen-antibody complexes deposit in vessel and cause complement
activation through the classical pathway. Main systemic manifestations include constitutional symptoms (fever,
fatigue), arthralgia and arthritis, pleuritis and obstructive lung disease, glomerulonephritis and interstitial nephritis,
pericarditis and cardiac valve disease, neuropathy, gastrointestinal symptoms (abdominal pain, nausea, vomiting,
diarrhea) and ocular disorders (episcleritis, uveitis, conjunctivitis). Skin biopsy is mandatory for diagnosis and shows
histopathologic features of leukocytoclastic vasculitis. Treatment is oriented by disease severity and include
antihistamines, steroids, dapsone, hydroxychloroquine and immunosuppressant drugs.
148 AUTOIMMUNE HEPATITIS AND THE OVERLAP SYNDROMES
X. Ma1
1Division
of Gastroenterology and Hepatology, Renji Hospital, Shanghai, China
Autoimmune hepatitis (AIH) is a chronic inflammation of the liver caused by an abnormal autoimmune reaction
against hepatocytes. The pathogenesis of AIH involves a loss of tolerance to hepatic self antigens in a susceptible
host. The diagnosis is based on histological abnormalities, and characteristic clinical and biochemical findings, which
include abnormal levels of serum globulins, and the presence of one or more characteristic autoantibodies such as
ANA, SMA, and anti-SLA. Interface hepatitis with abundant plasma cells in the infiltrate is characteristic histological
features of AIH. The simplified criteria have high sensitivity and specificity for diagnosis of AIH. The revised original
criteria have the complementary role to avoid the false negative diagnosis in atypical AIH patients.
Immunosuppressive treatment can attenuate hepatic inflammation, reverse fibrosis, and eventually improve the
patient's prognosis and life quality. Corticosteroids, either alone or in combination with azathioprine, are the standard
treatment of choice for AIH. Most patients with AIH show a good response to immunosuppressive treatment, although
patients with late-stage or severe disease are less likely to achieve remission.The simultaneous or successive
coexistence of AIH with primary biliary cirrhosis (PBC) in the same patient has been described as PBC-AIH overlap
syndrome. we found that there is a group of PBC-AIH patients who undergo a complete response to corticosteroids
and that this variant may be identified using the Paris criteria, especially when lowering the IgG threshold to 1.3
xULN, in conjunction with the simplified IAIHG scoring system.
149 ASSOCIATION OF ANTI-RIBOSOMAL P PROTEIN WITH RENAL MANIFESTATIONS IN SLE PATIENTS
S. Anis1, S. Sarfaraz1, E. Ahmed2, R. Muzaffar1
1Molecular
Diagnostics and Immunology, Sindh Institute of Urology and Transplantation, Karachi, Pakistan
Sindh Institute of Urology and Transplantation, Karachi, Pakistan
Background
2Nephrology,
Lupus nephritis (LN) occurs in 40-75% of systemic lupus erythmatosus (SLE) patients. Various autoantibodies have
been identified in SLE. Anti-ribosomal P protein (anti-P) antibodies are one of the marker antibodies in SLE.
Association of these antibodies with psychiatric or neurological symptoms has been reported. However their
involvement in LN is not very well known. We analyzed the presence of anti-P in LN patients.
Subjects and Methods
It is a descriptive and cross sectional study in which sera of 133 ANA positive SLE patients were investigated for the
presence of anti-P antibodies (from January 2010 to September 2012). Other autoantibodies tested were anti-dsDNA,
anti-Sm, anti-SS-A, anti-SS-B, anti-histones, anti-RNP and anti-PCNA antibodies. Clinical features and
histopathological classification of LN were also analyzed in these patients.
Data was analyzed using SPSS software version.12
Results
Anti-P antibodies were found in 15(11.5%) patients along with one or more other autoantibodies. Isolated anti-P
antibodiesÕ positivity was not found. Nine (60%) patients were positive for anti-dsDNA and eight (53%) for anti-SS-A
antibodies. Anti-P was mostly associated with class IV and class V lupus nephritis. The most common symptom was
pedal edema followed by arthralgia and oral ulcers. Three patients had history of seizures. Psychosis was not found
in any of these patients.
Conclusion
Presence of anti-P antibodies may add to the severity in SLE patients with LN. Therefore careful monitoring of these
patients may help in better patient management. We did not find an association of anti-P antibodies with psychiatric
involvement in our LN patients.
150 APRIL LEVELS STRONGLY CORRELATE WITH IL-17 IN SYSTEMIC LUPUS ERYTHEMATOSUS
G. Ostli-Eilertsen1, J. Nossent2
1Rheumatology,
University of Tromso, Tromso, Norway
Royal Darwin Hospital, Darwin, Australia
Introduction: Activation of self-reactive B cells plays an important part in Systemic Lupus Erythematosus (SLE). A
proliferation-inducing ligand (APRIL) and B cell activating factor (BAFF) are among the most specific B-cell
stimulators, but they activate B cells through unlike immunological pathways. We investigated the reciprocal
association between serum APRIL (s-APRIL) and serum BAFF (s-BAFF) in relation to clinical and immunological
findings in SLE patients.
2Rheumatology,
Methods: Cross sectional case-control study in 100 SLE patients (87% female, age 49 years, disease duration 12
years). S-APRIL and s-BAFF levels were measured by sandwich ELISA, compared with 31 healthy controls and
correlated with autoantibody, cytokine and clinical findings through nonparametric and multivariate regression
analyses.
Results: Both median s-APRIL (478 vs. 0 pg/ml, p=0.01) and s-BAFF (1720 vs. 0.9 pg/ml, p<0.001) were higher in
SLE patients than controls. Increased s-BAFF was observed in 86% of patients, but S-APRIL was increased only in
17% (p<0.01). S-APRIL correlated with s-BAFF in controls (p=0.04), but not in SLE (p=0.8). Increased S-APRIL was
strongly and independently associated with IL-17 activation (p<0.001), while increased s-BAFF levels was associated
with anti-nucleosome antibody presence (p=0.001), disease activity and organ damage. These were not associated
with increased s-APRIL.
Conclusions: Although both s-BAFF and s-APRIL levels are increased in SLE patients, they each reflect a different
immunological pathway. The strong association between s-APRIL and IL-17 suggests that IL-17 contributes to B cell
activation through s-APRIL but not BAFF.
151 A NOVEL IMMUNE-SPECIFIC THERAPY FOR MS-LIKE DISEASE BY A SYNTHETIC PROTEIN
ENCOMPASSING MULTI-APLS
N. Kaushansky1, L. Cohen1, A. Ben-Nun1
1Immunology,
The Weizmann Institute of Science, Rehovot, Israel
Among all approaches proposed for MS therapy, an approach that neutralizes only the pathogenic T cells reacting
against myelin, while leaving the innocent immune cells intact, is the ultimate goal in the immune-specific therapy for
MS. However, the multiplicity of primary target antigens, along side the dynamic nature of autoimmunity in MS,
whereby the specificity of anti-myelin pathogenic autoreactivities may shift or expand in the same patient with disease
progression, impose major difficulties in devising immune-specific therapy to MS. Among possible reasons for
therapeutic failure of clinical trial with MBP-APL, is the complexity of anti-myelin autoreactivities. To overcome this
multiplicity and complexity of pathogenic autoreactivities in MS, we have put forward the concept of concomitant
multi-antigen targeting as, a conceivably more effective approach to immunotherapy of MS.
We investigated such Òmulti-epitope-targetingÓ approach in EAE associated with multiple pathogenic
autoimmunities (ÒcomplexÓ EAE) in (C57Bl/6JxSJL/J)F1 mice via a multi-APL agent designated mMEP-APL, a
protein product of a synthetic gene designed to encode only characterized APLs of the defined encephalitogenic
epitopes of five myelin encephalitogenic proteins (MBP, MOG, PLP, MOBP and OSP).
mMEP-APL was found highly effective in suppressing and treatment of classical and ÒcomplexÓ EAE. We show that
systemic administration of mMEP-APL was associated with downregulation of Th1/Th17 cytokine secretion and
upregulation of TGF-b secretion together with induction of regulatory CD4+FoxP3+ cells.
Our data emphasize that a ÔÔmulti-epitope-targetingÕÕ strategy is required for effective immune-specific therapy of
organ-specific autoimmune diseases associated with complex and dynamic pathogenic autoimmunity, such as MS.
152 ESTABLISHMENT OF EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS IN CYNOMOLGUS
MONKEYS, A MODEL FOR MULTIPLE SCLEROSIS
J. Ni1, J. Liu1, J. Deng1, G. Cui1, L. Le1, S. Wang1, H. Zhang1
1Pharmacology,
Prisys Biotechnologies, Shanghai, China
he experimental autoimmune encephalomyelitis (EAE) model in rodents is widely used, however, the immune system
of rodents is quite different from humans, plus the genetic homogeneity and microbiological cleanness of the inbred
strains make the model system significantly apart from human multiple sclerosis (MS). To more closely model MS, we
established the EAE model in Cynomolgus monkeys. Brain white matter, spinal cord and MOG peptides were tested
for induction of EAE in monkeys. The typical disease related neurological symptoms, such as anorexia, weight loss,
ataxia, neck spasticity, and blindness, were observed two weeks after the immunization. A set of disease scoring
criteria was adopted to semi-quantitatively measure the disease severity. MRI examinations discovered the multiple
hyper-intense areas in the brain of the EAE monkeys at disease active stages, similar to those found in MS patients.
The findings of gross autopsy indicated multiple hemorrhagic lesions in the white matter of the affected monkey
brains. Both disease incidence and severity of the EAE were antigen-dose dependent. Interestingly, the disease
progression induced by brain white matter and spinal cord presented mixed types of the EAE episodes, while the
typical relapsing-remitting disease, representing the major type of MS in patients, was induced by MOG peptide.
Our results demonstrated the successful establishment of the EAE models and, for the first time, evidenced the
relapse-remitting EAE induced by MOG peptide in Cynomolgus monkeys. The models will help the more efficient
translation of the new treatments from concept to the patients for multiple sclerosis (MS) therapeutics.
Parallel Session: Parallel Session 10: Ankylosing Spondylitis and Immunoregulation
156 The role proinflammatory cytokines and SOCS in ankylosing spondylitis
C.T. Chou1, C.H. Chen2
1Division
of Allergy Immunology and Rheumatology, Veterans General Hospital Taipei, Taipei, Taiwan
of Allergy-Immunology-Rheumatology, Buddhist Tzu Chi General Hospital Taipei Branch, Taipei, Taiwan
Object: The high CRP in AS indicates the poor function and back deformity. In this study, we will measure CRP, ESR
and different cytokines and then correlate these data with clinical variables (BASDAI, BASFI, BASGI). Besides, we
also investigate the role of SOCS1 and SOCS3 in ankylosing spondylitis.
2Division
Method: Within one year, we have enrolled 181 AS patients. We measured different cytokines TNFα, IL1 β, IL6,
IL10, IL17 by ELISA. Simultaneously, we also evaluated BASDAI, BASFI, BASGI in AS patients. The expression
levels of SOCS1 and SOCS3 in mononuclear cells were measured by real-time PCR.
Results: When patients had normal ESR (<28), serum IL17α was significantly increased in CRP?1.0 vs CRP<1.0
(P=0.041). However, serum IL6 was significantly elevated in the group with CRP>5.0 when compared to CRP<5.0
(P=0.034). To correlate the different cytokines with the clinical variables, we demonstrated CRP levels had a positive
correlation with ESR (P<0.001), IL6 (P=0.037), BASFI (P=0.020), and BASGI (P=0.005). Except for CRP, ESR only
correlated with BASFI (P=0.024). IL6 was well-correlated with CRP, IL10 (P=0.011), TNFα (P=0.01), BASDAI
(P=0.033) and BASFI (P=0.018). SOCS3 expression in PMBC, T cells, and monocytes showed significant higher in
the AS patients than control (P = 0.025, 0.03, and 0.009 respectively).
Conclusion: The high CRP levels are significantly associated with IL6. Factors to affect the poor functional outcome
(BASFI) in AS patients are CRP and IL6. SOCS3 rather than SOCS1 may play a role the AS patients.
157 Spondyloarthritis and the role of Microbiota: lessons from SKG mice
R. Thomas1, L. Rehaume1, S. Hasnain2, S. Mondot3, H. Benham1, M.A. McGuckin2, M. Morrison3
1Diamantina
Institute, University of Queensland, Brisbane, Australia
Medical Research Institute, Mater Hospitals, Brisbane, Australia
3Animal Food and Health Sciences, Commonwealth Scientific and Industrial Research Organisation, Brisbane,
Australia
Spondyloarthropathies (SpA) including inflammatory arthritic, skin and bowel diseases share genetic susceptibility,
interleukin (IL)-23-dependence and involvement of the microbiota. However, it is unclear how the host genetic
background influences gut microbiota, and the microbiotaÕs relationship to organ inflammation in SpA. BALB/c
ZAP70W163C-mutant (SKG) mice develop SpA, including progressive peripheral and axial disease, unilateral uveitis,
CrohnÕs-like ileitis and psoriasis-like skin inflammation, triggered by microbial beta-glucan (curdlan) injection. Their
phenotype thus recapitulates human SpA with ileal inflammatory bowel disease (IBD).We found that microbiota
content and response to beta-glucan varied in mice with impaired T cell signal strength due to the SKG ZAP70W163C
mutation. Beta-glucan triggered acute inflammation and spondyloarthritis regardless of host genetic background or
microbiota. However, reconstitution of germ free SKG mice with a limited microbiota attenuated spondyloarthritis
severity. In contrast, ileal IL-23 expression, ER stress, lymph node IL-17 production, goblet cell loss and ileitis
development were microbiota-dependent. IBD but not peripheral arthritis was suppressed by microbiota transfer upon
co-housing SKG with BALB/c mice, as well as by TLR4 deficiency. These studies show that the interaction of genetic
background and host microbiota leads to an IL-23-dependent loss of mucosal function triggering ileitis in response to
beta-glucan. Moreover, genetic susceptibility to SpA can be modified by alteration of microbiota prior to exposure to
an infectious trigger.
2Mater
158 Update pathogenesis of spondyloarthritis
J. Wei1
1Department
of Allergy Immunology and Rheumatology, Chung Shan Medical University Hospital, Taichung, Taiwan
Spondyloarthritis (SpA), is a chronic inflammatory disease involving spine, peripheral joints and peri-articular soft
tissues. Up to 45% of patients may have extra-articular manifestations, such as uveitis, psoriasis, bowel inflammation
and nephropathy. The etiology is unknown, but probably related to genetics and immune dysregulation that was
triggered by environmental factors, mainly infection and trauma.
About 90% of AS patients are HLA-B27 positive. The positive rate of HLA-B27 is about 5% in normal Caucasian and
Chinese population. Thus, the odds ratio for AS in HLA-B27 carriers is greater than 100. HLA-B60 is weakly
associated with AS, with an odds ratio of 3.6. Recently, IL23R and ERAP1 have been reported in genome-wide
association studies.
Several hypothesis including molecular mimicry theory, arthritogenic peptide theory, ER stress theory, and HLA-B27
free-heavy-chain hypothesis have been proposed to explain the immunopathogenesis of AS and SpA. Klebsiella
pneumoniae (KP) was one of the most famous microorganism that involving in the pathogenesis of AS.
Biomechanical stress is another possible mechanism of causing inflammation in AS. HLA-B27 molecule was found to
be unfolded in the endoplasmic reticulum (ER), causing overloading ER stress and unfolded protein response (UPR).
Innate immunity and mucosal inflammation are major players in the pathogenesis of spondyloarthritis. IL-23 and IL-17
in Th17 axis are of interest in recent studies in the pathogenesis of SpA. The mechanism of new bone formation in
SpA is possibly due to sequelae of inflammation and fatty degeneration after inflammation, or activation of osteoblast
and bone morphogenetic proteins (BMP), though DKK-1, sclerostin or wnt pathway.
Parallel Session: Parallel Session 10: Ankylosing Spondylitis and Immunoregulation
160 INTERLEUKIN 10 DEFICIENCY SUPPRESSES THE PRODUCTION OF REGULATORY T CELL (TREG) –
DERIVED NEUROPHILLIN-1 (NRP-1) AND PROMOTES CD4+IFNG+ TH1 AND TH17 IMMUNITY
S. Wang1, C.K. Edwards III1, B. Guo1, L. Jiao1, Y. Wei1
1National
Key Laboratory of Biotherapy and Cancer Research, West China Hospital of Sichuan University, Chengdu,
China
Regulatory T cells (Treg) of the FoxP3+CD4+CD25+ phenotype play key roles in controlling reactivity to self-antigens
and the onset of autoimmunity. It has recently been demonstrated that Neurophillin-1 (Nrp-1) distinguishes natural
and inducible regulatory Treg subsets in vivo (Haribhai et al., 2011). In our laboratory, we have recently been
investigating the relationship between autoimmunity, anti-tumor, and Treg responses since the precise mechanisms
as to how Tregs infiltrate tumor tissues remain unclear. In the present studies, B16/F10 tumors were inoculated
subcutaneously into the left flank of either IL-10+/+ or IL10-/- C57BL/6 female mice, and tumor volume and weight were
monitored and recorded over a 21 day period. By day +15, there was a significant (p<0.05) decrease in tumor volume
in IL10-/- B16/F10 bearing mice compared to IL10+/+ B16/F10 bearing mice, and these differences remained highly
significant (p<0.001) through d+21. These tumor volume decreases correlated with significant (p<0.001) increases in
CD4+IFNγ+ and FoxP3+CD25+ Treg obtained from the spleen and lymph nodes from the IL10-/- B16/F10 versus
IL10+/+ B16/F10 mice at d+9 and d+15, respectively. Additionally, by d+15, intratumoral CD8+ T cells were significantly
(p<0.001) up regulated in the IL10-/- B16/F10 versus IL10+/+ B16/F10 mice, and these increases correlated with
significant (p<0.05) increases of spleen-derived CD4+IL17+ Th17 cells at both d+9 and d+15. Finally, we assessed the
d+9 expression of CD4+Nrp-1+ Treg cells in the tumor-draining lymph nodes (TDLN) and spleens of the IL10-/B16/F10 versus IL10+/+ B16/F10 mice. There were highly significant (p<0.001) increases in CD4+Nrp-1+ Treg
obtained from the LN of IL10-/- B16/F10 compared to IL10+/+ B16/F10 mice. Interestingly, we did not observe these
differences in CD4+Nrp-1+ Treg obtained from the spleens of IL10-/- B16/F10. In conclusion, our data indicate that
tumor growth in IL10-/- mice induces the specific activation of intratumoral tumor-specific CD8+ T cells, and at the
same time, it also elevates the expression of Treg in both the TDLN and spleen with Nrp-1 down-regulation in the
spleen. Our findings establish that IL-10 deficiency promotes Treg and CD4+IFNγ+ Th1 and Th17 cell populations
which act to suppress tumor growth in vivo.
Poster Session: Session 1: Animal models
P1-001 ESTABLISHMENT OF COLLAGEN INDUCED ARTHRITIS IN CYNOMOLGUS MONKEYS, A MODEL FOR
RHEUMATOID ARTHRITIS
J. NI1, J. Liu1, J. Deng1, G. Cui1, L. Le1, S. Wang1, H. Zhang1
1Pharmacology,
Prisys Biotechnologies, Shanghai, China
Since pharmaceutical compounds of new technologies, such as proteins, oligonucleotides or aptamers, etc.
frequently cross-react with the corresponding drug targets in non-human primates (NHP), it is apparently a more
efficient approach to evaluate their potential efficacy and pharmaceutical profile in a proper monkey disease models.
Collagen-induced arthritis (CIA) in monkeys has been reported, however, it is insufficient for characteristically
modeling rheumatoid arthritis (RA) due to the hampering of lower incidence and inconsistent disease. To establish a
reliable RA model, we characterized the disease progression following immunization of female Cynomolgus with
bovine type II collagen. In our studies, the overall incidence of individual arthritis reached 80% and the average
incidence of proximal interphalangeal (PIP) joint arthritis reached near 70%, significantly higher than those reported
previously. The PIP joints swelled approximately by 25% from the basal level in average in correspondence to the
increase of arthritic scores starting from the third week after first immunization. The pathological changes in joint bone
and cartilage were well supportive for the RA disease examined by radiopathology and bone histopathology. The
biomarker analysis of C-reactive protein (CRP) and ALP levels in peripheral blood showed a close correlation to the
arthritic incidence and severity along with the disease progress. The CIA monkeys also presented anemic symptoms,
a solid evidence for the similarity to RA with anemia of chronic disease (ACD) in the patients.
Our results demonstrated the establishment of CIA in Cynomolgus monkeys, with the characteristics of high similarity
to RA in human.
P1-002 NEUROPROTECTIVE CHANGES IN DEGENERATION-RELATED GENE EXPRESSION IN THE
SUBSTANTIA NIGRA FOLLOWING ACUPUNCTURE IN AN MPTP MOUSE MODEL OF PARKINSONISM
H. Jung1, S. Yeo1, Y. Hong1, Y. Choi1, S. Lim1
1Basic
Korean Medical Science, Kyung HeeUniversity, Seoul, Korea
ParkinsonÕs disease (PD) is a neurodegenerative disorder characterized by the death of dopaminegenerating cells
in the substantia nigra (SN). Acupuncture stimulation at GB34 and LR3 results in an enhanced survival of
dopaminergic neurons in the SN in Parkinsonism animal models. The SN is known to be an important player in the
neurological control of movement, and previous behavioral tests have found that acupuncture stimulation improves
motor dysfunction by approximately 87.7%. The present study investigated changes in gene expression profiles in the
SN region using a relatively chronic Parkinsonism model and a whole transcript array to identify genes that may be
related to the inhibitory effects of acupuncture on changes in gene expression following 1-methyl-4-phenyl-1,2,3,6tetrahydropyridine (MPTP) intoxication. In an MPTP model, acupuncture stimulation at GB34 and LR3 attenuated the
decrease in tyrosine hydroxylase in the SN region; stimulation at non-acupoints did not suppress this decrease.
GeneChip gene array analysis revealed that 22 (10 annotated genes:Cdh1, Itih2, Mpzl2, Rdh9, Serping1, Slc6a13,
Slc6a20a, Slc6a4, Tph2, and Ucma) probes that were up-regulated in MPTP animals relative to controls were
exclusively down-regulated by stimulation at acupoints but not non-acupoints. In addition, 17 (two annotated genes:
4921530L21Rik and Gm13931) probes that were down -regulated in MPTP animals compared to controls were
exclusively upregulated by stimulation at acupoints but not at non-acupoints. These findings indicate that the 39
probes (12 annotated genes) affected by MPTP and acupuncture may be responsible for the inhibitory
effects of acupuncture on degeneration-related gene expression in the SN following damage induced by MPTP
intoxication.
Poster Session: Session 1: Antiphospholipid Syndrome
P1-003 ANTIPHOSPHOLIPID-ANTIBODIES AND ANTIPHOSPHOLIPID-SYNDROME: WHAT IS THE
IMPORTANCE OF IGA-ANTI-BETA-2-GPI IN RENAL DISEASES?
S. Anis1, E. Ahmed2, R. Muzaffar1
1Molecular
Diagnostics and Immunology, Sindh Institute of Urology and Transplantation, Karachi, Pakistan
Sindh Institute of Urology and Transplantation, Karachi, Pakistan
AIM: To evaluate the significance of IgA-anti-beta-2-glycoproteinI-antibodies (IgA-anti-β2GPI) in the diagnosis of
antiphospholipid syndrome (APS) in patients with renal diseases.
2Nephrology,
Subjects and Methods: This is a retrospective study in which we have analyzed the prevalence of anti-β2GPI
isotypes and clinical features of APS in 22 patients. In all these patients, the test was repeated after 12 weeks for
confirmation of APS.
The tests for anti-β2GPI screening and its isotypes (IgG, IgM and IgA) detection were done by enzyme linked
immunosorbent assay using commercially available kits (Binding Site, Birmingham, UK).
The data was analyzed using SPSS software version 12 (Chicago, IL, USA).
Results: In 22 APS patients, IgA-anti-β2GPI was the most prevalent antibody compared to IgM and IgG isotypes
(91% vs. 45.5% and 18% respectively). Eleven (50%) of these patients had isolated IgA. Systemic lupus
erythmatosus was present in 12 patients with mostly IgA isotypes (10; 83%). The IgA-antiβ2GPI titers were moderate
to high in 12 patients on initial testing and remained high in 10 patients on repeat testing.
Various manifestations of APS in these patients were recurrent thrombosis, livedo reticularis, digital infarction,
pregnancy complications, arterio-venous-fistula-formation (AVF) failures and renal APS. Interestingly IgA isotypes
were found in 100% of patients with pregnancy morbidity, AVF failures and renal APS while IgG-anti-β2GPI were
absent in these patients.
Conclusions: Our results show that IgA-anti-β2GPI has a significant role in diagnosing APS in our patient
population. A larger multicentric prospective study is required to elucidate the importance of IgA-anti-β2GPI testing to
diagnose APS.
P1-004 PREVALENCE OF CLASSICAL ANTIPHOSPHOLIPID ANTIBODIES IN THYROID DISEASES
M. Barzilay1, Y. Sherer2
1Medicine
F, Tel Aviv Sourasky Medical Center, Tel-Aviv, Israel
Management, Tel Aviv Sourasky Medical Center, Tel-Aviv, Israel
Background: Antiphospholipid syndrome (APS) is associated with thrombosis and pregnancy loss in the presence of
antiphospholipid antibodies (aPL), mainly anticardiolipin (aCL), anti-beta2-glycoprotein I (anti-β2GPI) and lupus
anticoagulant (LAC). aPL antibodies can also be found in other circumstances in which they are not associated with
clinical APS criteria, such as infections and other autoimmune diseases.
2Hospital
Objective: To evaluate the prevalence and levels of aCL, anti-β2GPI and CL-β2GPI complex antibodies in a cohort
of patients with thyroid diseases.
Methods: Sera from 68 patients with Graves' disease, 50 patients with Hashimoto thyroiditis, and 237 healthy
subjects were analyzed by the BioPlex 2200 for aCL (IgG, IgM), anti-β2GPI (IgG, IgM) and CL-β2GPI (IgM, IgG)
antibodies.
Results:
APL were found positive and at higher levels than healthy controls only in Graves disease, whereas no significant
difference was found among Hashimoto thyroiditis patients. IgM anti-CL-B2GPI, IgG anti-B2GPI and IgG anti-CLB2GPI were found at higher levels among Graves' disease compared with healthy subjects. In addition, IgM anti-CLB2GPI and IgG anti-B2GPI were more prevalent among Graves patients compared with controls.
Conclusions:
Sub-groups of patients having Graves disease and positive for aPL might be at risk for thromboembolic events as
should be further studied.
P1-005 SEVERE CUTANEOUS MANIFESTATIONS IN A PATIENT WITH ANTIPHOSPHOLIPID SYNDROME
I. Florea1, I. Vais2, C. Tutunaru2, N. Florea3, L. Predoi2
1dermatology,
emergency clinical hospital, craiova, Romania
emergency clinical hospital, Craiova, Romania
3Radiology, emergency clinical hospital, Craiova, Romania
Antiphospholipid syndrome is the most common cause of thrombophilia. It's an autoimmune disease characterized by
the association of multiple thrombosis in any vascular level (both arterial and venous). Our aim is to confirm the
diagnosis of antiphospholipid syndrome.
2Dermatology,
We will present the case of a 17 year-old male patient, hospitalized for the presence of multiple ulcers with a chronic
venous insufficiency history, with development of collateral circulation at the abdominal and chest wall level (jellyfish
head) and giant veins on the legs. The patient was uncooperative, initially claiming that the lesions are due to the dog
bites. He had undergone right nephrectomy two years before in the context of nephritic syndrome and the presence
of renal vein thrombosis.
Postoperative, he showed signs of thrombosis at the inferior cava vein and iliac veins, with the development of
collateral circulation. Suspecting a retroperitoneal process, he underwent extensive laboratory investigations. MRI
confirmed the existence of a retroperitoneal lesion process that reduces the size of cava vein in its lower section,
extended to the iliac veins axes. Biological samples showed positive anticardiolipin antibodies, positive lupus
anticoagulant, deficiency of protein C and antitrombine III. Because of these changes we are entitled take into
account antiphospholipid syndrome with severe skin manifestations and because of the patientÕs age we are
confronted with hereditary thrombophilia.
The patient was transferred to the vascular surgery service.
The particularity of the case lies in the fact that such pathology is encountered in women between the ages of 30 and
40 years.
Poster Session: Session 1: Autoimmunity - other
P1-006 COMPARATIVE ANALYSIS OF TWO LUMINEX-BASED SYSTEMS FOR DETECTION OF ANTINUCLEAR ANTIBODIES
M. Cheng1, C. Lee1, J. Lin1
1Laboratory
department, Changhua Christian Hospital, Changhua, Taiwan
Background: The Luminex-based system is now a popular tool for simultaneous detection of a panel of anti-nuclear
antibodies (ANA). However, equivocal results are not uncommon with uncertain reasons. The aim of this study is to
compare two Luminex-based systems for ANA testing.
Materials and Methods: A total of 132 clinical sera submitted for ANA tests were analyzed on both AtheNA MultiLyte system (semi-automated) and Bioplex 2200 system (fully automated). ANA including anti-SSA, SSB, Sm, RNP,
Scl70 and Jo1 were compared for concordance between systems after exclusion of equivocal results. Sera with any
disconcordant ANA were re-run and subsequently tested in duplicate using a reference method based on enzymelinked immunosorbent assay (ELISA) for accuracy assessment.
Results: The concordance rates (kappa coefficient) between two Luminex systems were 91% (0.81) for SSA
(n=125), 88.0% (0.64) for SSB (n=125), 96.0% (0.72) for Sm (n=130), 91% (0.66) for RNP (n=129), 96% (0.43) for
Jo1 (n=131) and 92% (0.51) for Scl70 (n=130). Disconcordance occurred in 22 sera (16.7%). Among them, the
duplicate reproducibility in terms of interpretation was 13.6% (3 sera) by AtheNA and 86.4% (19 sera) by BioPlex.
Besides, the analytical accuracy for AtheNA and BioPlex was found to be 27.3% and 72.7% for SSA (n=11), 30.8%
and 69.2% for SSB (n=13), 80.0% and 20.0% for Sm, 36.4% and 72.7% for RNP, 20.0% and 80.0% for Jo1, and
10.0% and 90.0% for Scl70, respectively.
Conclusion: The performance of BioPlex in ANA testing seems superior to AtheNa in terms of discrimination power,
duplicate reproducibility and accuracy.
P1-007 A CASE REPORT OF EVANS SYNDROME
D. Fetarayani1, G. Soegiarto1, M.P. Sedana1
1Internal
Medicine, Medical Faculty of Airlangga University- Dr. Soetomo General Hospital, Surabaya, Indonesia
INTRODUCTION : Evans syndrome (ES) is an autoimmune disorder characterized by the simultaneous or sequential
development of autoimmune hemolytic anemia (AIHA) and immune thrombocytopenia (ITP). ES is a rare condition,
because it is diagnosed in only 0.8% to 3.7% of all patients with either ITP or AIHA at onset.
CASE : A 30-year-old male had complaint of weakness, nausea, vomiting, decrease of appetite of seven days
duration. Palpitations and dyspnea on exertion since two days prior to admission. He had been diagnosed as
hemolytic anemia since seven months before, but not routinely visit the out-patient clinic. There were history of
recurrent epistaxis and gum bleeding. On examination, the patient was severely pale and slight icteric on sclera.
There were splenomegaly, ptechiae/purpura on both upper and lower extremities. Laboratory results revealed Hb 2.4
g/dL, HCT 6.4%, leucocyte 12,200/μL, thrombocyte 8,000/μL, reticulocyte 6.5%, ESR 30 mm/1 hour. Peripheral blood
smear revealed hypochromic with anisopoikilocytosis, normoblast, and thrombocytopenia. Serum bilirubin was 3.21
mg/dL with a direct level of 0.76 mg/dL. Direct Coombs test was positive. The other causes of thrombocytopenia and
hemolytic anemia were excluded. Serum IgM, IgG, and IgA were normal. Bone marrow aspiration revealed
normocellular marrow, Myeloid:Erythroid ratio was 3:1, with an increase of thrombopoetic system. The patient started
on a course of corticosteroids and other supportive treatment. Two weeks after admission, he began to stabilize. He
was discharged with Hb 8.96 g/dL, reticulocyte 3.8%, and thrombocyte 270,000/μL on methylprednisolon which was
then tappered.
Keywords : Evans syndrome, autoimmune hemolytic anemia, immune thrombocytopenia
P1-008 CHANGES OF T REGULATORY CELLS IN PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY
DISEASE
L. Jurgauskiene1, V. Sileikiene2, E. Danila2, R. Malickaite1
1Laboratory
of Immunology, Vilnius University, Vilnius, Lithuania
of Pulmonology and Allergology, Vilnius University, Vilnius, Lithuania
Background and aims Chronic obstructive pulmonary disease (COPD) is characterized by progressive airway
obstruction and airway inflammation. What cells and how they are causing this ongoing chronic inflammation in
airways are still unknown. Researchers suggest that T regulatory cells may play a role in COPD as it is thought that
COPD is an autoimmune disease.
2Center
Methods We analysed T regulatory cells CD3+CD4+CD25+CD127dim and CD3+CD4+CD25bright in peripheral blood
samples of patients (pts) with different COPD stages. 60 pts (mean age * 53 ± 15 yrs), where I gr. * 20 pts, healthy
people, II gr. * 20 pts with mild and moderate COPD (FEV1/FVC < 0.7, FEV1 80% and 50% FEV1 80%), III gr. * 20 pts
with severe and very severe COPD (FEV1/FVC < 0.7, 30% FEV1 < 50% and FEV1 < 30% or FEV1 < 50% analysed.
Peripheral blood samples were analyzed by flow cytometry (BD FACSCalibur) using four color monoclonal antibodies
to cell surface antigens.
Results Our study showed no significant differences in number of CD3 +CD4+ CD25+CD127dim T cells (1.1±0.7;
0.9±0.8; 0.8±0.6 respectively), but we noticed that there is a tendency that these cells are decreasing with severe of
COPD. While the number of CD3+CD4+CD25bright T cells was significant (p < 0.05) decreased in III group comparing
with other two groups (3.2±1.2; 3.4±1.3; 2.5±1.3 respectively).
Conclusions According to our assay results we can say that CD3 +CD4+CD25bright and CD3+CD4+ CD25+CD127dim T
cells are playing a role in COPD inflammation and we can speculate that COPD could be an autoimmune disease.
P1-009 MYOID CELLS PLAY A KEY ROLE IN MYASTHENIA GRAVIS AND HYPERPLASIA
I. Kamo1, H. Tomoyasu2, A. Kikuchi3
1Bio
Lab., System Instruments Co. Ltd., Sagamihara, Japan
Omori Red Cross Hospital, Omori, Japan
3Department of Physics and Mathematics, College of Science and Engineering Aoyama Gakuin University,
Sagamihara, Japan
Autoimmune disease myasthenia gravis is associated as much as ~80% with thymic hyperplasia where many B-cells
and IL-2 producers, but virtually no IL-4 or IFN-γ producers, are proliferating in an autonomous and/or cytokine
dependent fashion. Increased numbers of precursor myoid cells were also detected here with specific antibodies
against new 80-kDa and 100-kDa haemopoietic factors purified from rat myoid cells. They can distinguish hyperplasia
from thymoma. The myoid cell conditioned medium, containing IL-1, IL-6 and IL-7 in addition to the above factors,
induced antigen independent IL-2 production from thymocytes and stimulated the proliferation of lymphocytes and
monocytes. IL-2, the only lymphokine produced here, is not the B-cell proper. IL-2 and myoid cell factors are likely to
be overproduced according to the increases of their producers, thus forming a new B-cell stimulatory cytokine
network. We tested this possibility by constituting mimic hyperplastic thymic conditions: nude mouse splenocytes that
include functional B-cells and antigen presenting cells, but essentially lack mature Th1- and Th2-cells, were cultured
in the presence of IL-2 and myoid cell factors. When combined, they indeed induced antigen specific B-cell
responses in a synergistic fashion, whereas each factor alone poor responses. This cocktail also induced
immunoglobulin class switches from IgM to IgG subclasses. We believe that myoid cells play an important role in
forming lympho-proliferative cytokine network (hyperplasia) and present their antigens to the corresponding B-cells,
thus myoid-muscle antigen specific B-cells are selectively activated here (myasthenia gravis). The mechanism of
myoid cell development remained to be elucidated.
2Respiratory,
P1-010 PROTEOMIC PROFILING OF AUTOIMMUNITY REPERTOIRES WITHIN MULTIPLE SCLEROSIS
UTILIZING ANTIGEN ARRAYS AND HIGH-DENSITY PEPTIDE MICROARRAYS
B. Ayoglu1, A. Zandian1, A. Häggmark1, B. Forsström1, M. Khademi2, T. Olsson2, M. Uhlén1, J.M. Schwenk1, P.
Nilsson1
1SciLifeLab
Stockholm, KTH - Royal Institute of Technology, Solna, Sweden
of Clinical Neuroscience, Karolinska Institute, Solna, Sweden
Broad scaled screening and profiling of autoantibody repertoires for the discovery of new autoimmune targets
requires large antigen collections representing a significant part of the human protein coding genes. We have here
utilized both planar and bead based protein arrays containing large sets of protein fragments from the Human Protein
Atlas (www.proteinatlas.org) as well as ultra-high-density peptide microarrays for the profiling of plasma and CSF
samples within multiple sclerosis.
2Department
More than 11.000 protein fragments on planar microarrays, representing more than one third of the human protein
coding genes, were initially screened with 90 plasma samples and followed up by a verification phase with 380
samples on 380 targets on suspension bead arrays. The outcome was 51 potential autoimmunity targets
differentiating between subgroups of multiple sclerosis. 1 Follow up studies is initiated where these selected candidate
targets together with other suggested autoimmune targets are profiled with 4000 multiple sclerosis related samples.
Ultra-high density peptide microarrays were also utilized for screening for novel autoimmunity targets. The peptide
arrays were designed with either 2 million peptides with whole proteome coverage with 6 out of the 12 amino acids
overlap or 150.000 peptides representing a selected set of targets and then also enabling a higher resolution with 11
amino acids in overlap. The whole proteome arrays revealed a frequently repeated and specific part of a receptor
domain as a potential novel autoimmunity target in multiple sclerosis.
1
Ayoglu etal, Autoantibody profiling in multiple sclerosis using arrays of human protein fragments. Molecular &
Cellular Proteomics June 2013
P1-011 GAMMADELTA T CELLS INFILTRATION AND PROINFLAMMATORY CYTOKINES PRODUCTION IN
PATIENTS OF TAKAYASU ARTERITIS
L. Pan1, Y.M. Liu2, T. Wang1
1Rheumatology
and Immunology, An Zhen Hospital Affiliated to Capital Medical University, Beijing, China
Surgery, An Zhen Hospital Affiliated to Capital Medical University, Beijing, China
Background and aims: Series of lymphocytes are recruited and and may produce cytokines which involved in the
lesions of the aortic wall to participate in the pathogenesis of TA, but the mechanism is not clear.
2Cardiac
Mathods: Blood samples will be collected from 5 TA patients and 5 healthy controls; tissues of arota will be taken
from a TA patient by surgery. Cytometric Bead Array Flex Set was used to analyze the proinflammatory cytokines
production in serum and immunohistochemical staining to detect T cells infiltration in tissues of arota of TA patients.
Results: Our study found that CD8+ T cells and gammadelta T cells infiltration in aorta tissue of TA patient, CD8:
HLA - I and NKG2D: MICA antigens presented process were showed in aorta tissue of TA patient (Fig 1). Serum
inflammatory cytokine IL-8 and IL-9 levels were significantly higher in TA patients than that of normal control group
(P<0.05), IL-6 and TNF-α levels also tended to increased.
Conclusion: Gammadelta T cells activated by accepted antigen through CD8: HLA - I and NKG2D: MICA pathway,
and then infiltrate in the aortic wall and produce inflammatory cytokines such as IL-8, IL-9, IL-6 and TNF-α to
participate in the pathgenesis of TA.
P1-012 CHILDHOOD PRIMARY ANGIITIS OF THE CENTRAL NERVOUS SYSTEM: A REPORT OF TWO CASES
C. Tan1, C.B. Bernal1, S.V. Navarra1
1Rheumatology,
University of Santo Tomas Hospital, Manila, Philippines
Background and Objective: Primary angiitis of the central nervous system (PACNS) is a rare disease wherein the
pathogenesis is not well understood. It has an estimated annual incidence of 2.4 cases per 1 million person-years.
Middle aged men are more often affected with a median age of approximately 50 years. On literature review, Lindsley
et al (2005) reported that there are fewer than 50 cases in children that were published. This paper aims to present 2
pediatric cases of PACNS seen in UST Hospital.
Case 1: An 11 year old, female presented with sudden onset of drowsiness with left sided weakness. Laboratory
work-ups were unremarkable, ANA panel was negative, blood cultures revealed no growth, MRI revealed subacute
infarct in the right MCA distribution and MRA revealed stenosis of the anterior cerebral artery and right middle
cerebral artery.
Case 2: An 11 year old male presented with chronic insidious headache for four years. He was initially managed as a
case of migraine and vertigo, blood examinations were unremarkable, however on further work-ups, MRA revealed
irregularities on the left posterior cerebral artery consistent with vasculitis.
Conclusion: There are no controlled trials on the treatment of PACNS and therapy is based on principles
extrapolated from systemic vasculitis. Both patients were given glucocorticoids and cyclophosphamide which
apparently arrested progression, although residual neurologic deficit in one patient additionally required intensive
physiotherapy.
Funding: Lupus-Inspired Advocacy Project of Rheumatology Educational Trust Foundation Inc.
P1-013 BIOENERGETICS OF THE SPINAL CORD TISSUE IN EXPERIMENTAL ALLERGIC ENCEPHALITIS OF
RATS
M. Al Shamsi1, A. Shahin1, A.K. Souid2, E.P.K. Mensah-Brown3
1Microbiology
& Immunology, United Arab Emirates University, Al-Ain, United Arab Emirates
United Arab Emirates University, Al-Ain, United Arab Emirates
3Anatomy, United Arab Emirates University, Al-Ain, United Arab Emirates
Introduction: Experimental autoimmune encephalomyelitis (EAE). a rodent model of relapsing/remitting multiple
sclerosis (MS), is a CD4+ T cell-mediated inflammatory disease of the central nervous system (CNS). Demyelination
and axonal damage, characteristics of EAE have been attributed also to dysregulation in mitochondrial function.
Objective: To demonstrate that cellular respiration or bioenergetics of the CNS could predict prognosis of EAE.
Aims of study:To determine whether functional state of neuronal mitochondria after disease induction was linked to
severity of EAE.
Methods:Susceptible Dark Agouti (DA) and resistant Albino Oxford (AO) rats were monitored for induction, height of
and recovery from neurologic disease after immunization with myelin basic protein and complete FreundÕs adjuvant.
Sections of the lumbar region of the spinal cord were analyzed for cellular respiration by measuring oxygen and ATP
content using a phosphorescence analyzer. Caspase activity, histological signs of inflammation and cell damage
were also determined by HPLC, FACS, cytochrome C oxidase immunohistochemistry, light and electron microscopy.
Result: Our results revealed preserved respiration and ATP content of the CNS throughout the course of disease.
This was associated with intact and normal mitochondria. Transient caspase-3 activity observed and confirmed by
HPLC, FACS and caspase-3 immunohistochemistry in DA rats at the height of disease disappeared on recovery. In
addition, evidence of demyelination was unremarkable.
Conclusion:Our results suggest that recovery from EAE might be attributable to the preserved bioenergetics in this
initial wave of the disease.
2Paediatrics,
P1-014 MIXED CONNECTIVE TISSUE DISEASE WITH MANIFESTATION OF THROMBOANGIITIS OBLITERANS
D. Mulya1, N. Sukmana2
1Internal
Medicine, Gadjah Mada University, Yogyakarta, Indonesia
and Clinical Immunology, University of Indonesia, Jakarta, Indonesia
Introduction
Thromboangiitis obliterans (TAO) is a recuring progressive inflamation and thrombosis of small and medium arteries
and vein of the hand and feet. Risk factor associate with thromboangiitis obliteran is a heavy smoker.Upper extremity
digital ulceration due to ischemia also occur as a complication of Mised Connective Tissue Disease (MCTD) and
vasculitis. We report 53 years old man with MCTD and amputated finger diagnose as thromboangiitis obliteran.
Case Presentationblackish
A 53 years old man complained of swelling and blackish at end of his hand since two month ago. Two weeks before
admittance, tip of his finger amputated by itself. Patient also had diabetes and smoking since the age of twenty. On
examination amputated finger was found at the right hand from digiti I-V. Swelling was also found in his right palm.
Glucose level was 360 mg/dl. ANA test 1/1000. RNP/sm ++, CRP level was 41,2, ESR was 260, ACA IgG and IgM
negative. There was a plaque and stenosis 50-75% in his brachial artery and his left and right ulnar radialis with
minimal blood flow in his right hand. Patients was diagnose as MCTD and TAO. Patients was adviced to stop
smoking and treated with cyclophosphamid 500 mg every two weeks, methylprednisolon 60 mg tappering off,
cilostazol 50 mg bd, vitamin D, folic acid and insulin.
Conclusion
We report 53 years old man with TAO and MCTD. Autoimmune disease and comorbidities in patient brings challenge
in therapy and futher prognosis.
2Allergy
P1-015 SOCS3 METHYLATION IN SYNERGY WITH REG? OVEREXPRESSION TO ACCELERATE
INFLAMMATION-LINKED PANCREATIC CARCINOGENESIS
M. Xiang1, J. Wang2
1College
of Pharmacy, Tongji Medical College, Wuhan, China
of Pharmacology, Medical CollegeWuhan University of Science and Technology, Wuhan, China
Background and aims: We evaluated the effect of SOCS3 methylation in synergy with RegIII overexpression in the
inflammatory environment to accelerate pancreatic carcinogenesis.
2Dept
Methods: Real-time PCR and methylation-specific PCR were carried out in tumor material from 36 pancreatic cancer
(PaC) patients and 4 PaC cell lines BxPC-3, Mia Paca-2, PANC-1, SW1990 to assess RegIIImRNA expression and
the methylation status of the SOCS3 CpG island respectively. SOCS3 gene was transfected into exogenous RegIIIinduced PaC cell lines and silenced using siRNA in normal pancreatic epithelial cell line HPDE6C7 in vitro to assess
the status of SOCS3 interferes with exogenous RegIII in cell growth. Results: Clinical observations showed that
SOCS3 methylation and RegIII overexpression have independent correlation with the clinicopathological factors
including inflammation history, maximal tumor size, tumor differentiation and TNM stage. And RegIII mRNA
expression in the samples with SOCS3 methylation was significantly higher than those with SOCS3 unmethylation.
And we identified aberrant SOCS3 methylation in PaC cell lines associated with transcriptional downregulation.
Treatment of the demethylating agent 5-aza-2'-deoxycytidine restored SOCS3 expression of SW1990 cells.
Overexpression of SOCS3 in PaC cells inhibited the proliferation-promoting effect of exogenous RegIII. Conversely,
siRNA-mediated knockdown of SOCS3 enhanced RegIII-induced cell proliferation of HPDE6C7 cells.
Conclusions: SOCS3 methylation may synergize with RegIII overexpression to correlate with accelerated onset of
inflammation-linked pancreatic carcinogenesis, suggesting SOCS3 is a promising molecular target for the control of
RegIII-stimulated pancreatic carcinogenesis.
P1-016 ARTHRITIS ATTACK DUE TO MEAT CONSUMPTION
A. Kutlu1, S. Ozturk1, O. Taskapan2, Y. Onem3, M.Z. Kiralp4, L. Ozcakar5
1Allergy
and ?mmunology, GATA Haydarpa?a training Hospital, Istanbul, Turkey
and ?mmunology, Yeditepe University, Istanbul, Turkey
3Internal medicine, GATA Haydarpa?a training Hospital, Istanbul, Turkey
4physical medicine and rehabilitation, GATA Haydarpa?a training Hospital, Istanbul, Turkey
5physical medicine and rehabilitation, Hacettepe University, Ankara, Turkey
32-year-old woman was seen for a possible diagnosis of food allergy. She had a diagnosis of rheumatoid arthritis and
had been monitored for the last 8 years . On detailed questioning, she reported having several arthritis episodes
when she ate a diet rich in animal food (eggs, milk, and meat). The episodes started 6-10 hours after ingestion of
animal protein (most severe with meat) and lasted 2-3 days.
She was free of arthritic episodes when she stopped ingesting animal products. She did not notice any symptoms
when she followed a vegetarian diet composed of plant proteins (peas, lentils, beans, etc). Gouty arthritis was ruled
out based on clinical signs and several normal uric acid measurements. When the patient underwent skin testing and
oral diet challenge, she reported that she had ingested meat the night before the test in order to demonstrate her
complaint of arthritis. During physical examination on the day of testing, swelling, erythema, and tenderness were
detected on the left knee joint. Allergy skin testing was negative for all types of aforementioned food. Allergenspecific, cutaneous lymphocyte antigenpositiveT cells have been shown to play a role in delayed eczematous skin
reactions in atopic dermatitis.4 We believethat a similar type of organ-specific type IV immunologic reaction might
have taken place in our patientÕs joints. It has been mentioned that food-related problems of RA patients might
reflect an adverse additive effect of multiple hypersensitivity reactions mediated by immune complexes promoting
autoimmune reactions in the joints
2Allergy
P1-017 PULMONARY INVOLVEMENT IN POLYMYOSITIS AND DERMATOMYOSITIS
A. Zoto1, H. Hafizi2, T. Backa1, R. Osmenaj3, A. Harxhi4, Z. Ylli5
1Department
of Rheumatology, University Hospital Center “Mother Theresa”, Tirana, Albania
of Lung Diseases, University Hospital of Lung Diseases “Shefqet Ndroqi”, Tirana, Albania
3Department of Radiology, University Hospital Center “Mother Theresa”, Tirana, Albania
4Department of Infectious Disease, University Hospital Center “Mother Theresa”, Tirana, Albania
5Department of Immunology, University Hospital Center “Mother Theresa”, Tirana, Albania
Background and aims: Polymyositis and Dermatomyositis are systemic inflammatory diseases that affect skeletal
muscles and other internal organs such as lung. Pulmonary involvement in polymyositis and dermatomyositis
includes interstitial lung disease, aspiration pneumonia, respiratory muscle weakness. The aims of this study was the
identification of pulmonary involvement in patients with polymyositis and dermatomyositis, their assessment in
relation to alterations of immunological examinations
2Department
Methods: This is a cohort prospective study that analyzed 51 patients with polymyositis and dermatomyositis. The
laboratory tests requested for the patients such as anti-nuclear antibody, anti-histidyl t ribonucleic acid synthetase
antibody. Pulmonary high resolution computed tomography and pulmonary function test were performed for the
patients.
Results: Interstitial lung disease was found in 15 (29.4%) patients, aspiration pneumonia in 6 (11.8%) patients and
respiratory insufficiency in 1 (2%) patients. Restrictive ventilator insufficiency was observed in 13 (25.5%) patients.
Pulmonary injuries in the group of patients with abnormal immunological laboratory tests are found in 17 (63%) of
patients, while in the other group of patients who do not have positive anti-nuclear antibody and anti-histidyl t
ribonucleic acid synthetase antibody, pulmonary injuries are found in 5 (20.8%) of patients.
Conclusions: Pulmonary manifestations are common in polymyositis and dermatomyositis. These injuries are
anatomical and functional. Immunological alterations are important factors in pulmonary injuries.
Key words: myositis, lung disease, antinuclear antibody.
P1-018 ANTI-NMDA RECEPTOR ENCEPHALITIS: A CASE
N. Akgun1, D. Erdogan Ari1, C. Karip1, E. Gözke2, C. Agalar3
1Anesthesiology
and Reanimation, Fatih Sultan Mehmet Education and Research Hospital, Istanbul, Turkey
Fatih Sultan Mehmet Education and Research Hospital, Istanbul, Turkey
3Infection Diseases and Clinical Microbiology, Fatih Sultan Mehmet Education and Research Hospital, Istanbul,
Turkey
BACKGROUND: A case of autoimmune encephalitis related to anti-NMDA antibody is discussed. CASE REPORT:
Twenty-two years old female patient suffering from abdominal pain and nausea admitted to emergency service three
times within 45 days. Headach accompanied by apathy, impaired cooperation was present at third admission. Her
mother death was related to an encephalitis with unknown ethiology. Mild alteration in the intensity of left
parahippocampal girus was detected on cranial MRI. CSF analysis showed the presence of PMN leukocytes
(80/mm3). The patient was hospitalized for the treatment of encephalitis. The result of PCR testing for Herpes virus
was negative. Autoantibody scanning showed the presence of plasma antibodies against NMDA receptors.
Metilprednisolone 1000 mg/day was added to drug therapy. Following the deterioration of neurological status
(GCS:7), she was accepted to ICU where intravenous immunoglobulin 0.4 mg/kg/day was administered. PET-CT
showed no malignacy. Conscious but uncooperative patient received five sessions of plasmapheresis using Asai
Kasei plasma exchange kit followed by immunoadsorption using the same companyÕs immunoadsorption colon.
Following fifth session, immunoabsorption was discontinued due to the onset of severe fever. Two days later,
ooferectomy was performed at postmortem first hour in order to detect any undiagnosed overian teratoma.
Histopathological evaluation revealed no pathological findings. DISCUSSION: Anti-NMDA receptor antibody-related
encephalitis in young women may be associated with ovarian teratoma (1). It can also be encountered in the absence
of malignancy, in both gender, whatever the age of the patient (1). In a case series of 400 patients, exitus was
observed in all of the 15 patients accepted to ICU (2).REFERENCES: 1- Neurol Clin Pract. 2012 Sep;2(3):215-223,
2-QJM. 2011 Nov;104(11):921-31
2Neurology,
P1-019 CRYOPRESERVATION OF VITAMIN D3 [1,25(OH)2D3]-TREATED DENDRITIC CELLS : A CRUCIAL
STEP TOWARDS CLINICAL APPLICATION OF TOLEROGENIC DC VACCINATION
W.P. Lee1, B. Willekens2, Z.N. Berneman1, N. Cools1
1Vaxinfectio
Laboratory of Experimental Hematology, University of Antwerp, Antwerp, Belgium
of Neurology, Antwerp University Hospital, Edegem, Belgium
While emerging evidence indicates that dendritic cells (DC) play a central role in the initiation and progression of
multiple sclerosis (MS), their modulation with immunoregulatory agents provides prospect as immunosuppressive
therapy. The aim of this study is to investigate the capacity of tolerogenic DC (tolDC) to restore or modulate
pathogenic responses in MS. Our observations reveal that 1,25-dihydroxyvitamin D3 (10-5M) treatment of DC results
in a maturation-resistant phenotype and anti-inflammatory cytokine profile as compared to conventional IL-4-treatedDC. Importantly, we demonstrate the ability of vitD3-treated-DC loaded with myelin-derived peptides to induce stable
antigen-specific hyporesponsiveness in myelin-reactive T-cells derived from healthy volunteers and MS-patients.
Indeed, T-cells were still responsive to an unrelated antigen, such as the cytomegalovirus pp65 peptide, indicative of
antigen-specific T-cell hyporesponsiveness. Moreover, induced hyporesponsiveness was stable, since these T-cells
did not reactivate upon stimulation with fully mature conventional DC. Finally, we developed a cryopreservation
protocol which yielded 78% recovery and 75% viability of immature vitD 3-treated-DC following a freeze-thaw cycle.
Furthermore, no differences regarding the expression of typical DC membrane-markers and T-cell stimulatory
function could be observed before and after cryopreservation.
2Division
In conclusion, we demonstrate, to our knowledge for the first time, the feasibility of cryopreservation of tolDC. In this
perspective, our results contribute to potential large scale production and preservation of tolDC and further underline
their possible clinical applicability in order to correct the immunological imbalance in MS.
P1-020 DIAGNOSIS CHALLENGE IN A SEVERE CASE OF ACRODERMATITIS CONTINUA OF HALLOPEAU
I. Florea1, I. Vais1, C. Tutunaru1, L. Predoi1, N. Florea2
1dermatology,
emergency clinical hospital, craiova, Romania
emergency clinical hospital, craiova, Romania
Acrodermatitis continua of Hallopeau is a rare variant of pustular psoriasis with chronic evolution that tends to be
resistant to topical and systemic treatment.
2Radiology,
We present a case of a 63 year-old male, who checked in repeatedly for erythematous and pustular quasigeneralized eruption with severe involvement of the fingers and toes. The patches are sharply demarcated from
healthy skin with multiple intraepidermic pustules that break easily leaving behind a bright red, glazed, slightly
atrophic epiderma that in evolution is covered by crusts. In addition, the actual episode is accompanied by functional
limitation of the hands and feet due to extension of the disease. Also the nail examination revealed severe
onychodystrophy, onycholysis and anonychia. The onset of the disease was in January 2011 with small palmoplantar
pustules either isolated either grouped forming plycyclic lakes of pus.
The tests revealed hyperleukocytosis, hyperglycemia, increased ESR and elevated liver enzymes and the
bacteriologic examination was negative.
The histopathological examination confirmed the diagnosis of acrodermatitis continua of Hallopeau and the x-ray
showed diffuse demineralization of the distal phalanx, arthropathy of the interphalangeal joints and diffuse
osteoporosis due to postinfectious left calcaneal geostoza.
Because the corticotherapy has proven effective only in the short-term we started systemic therapy with retinoids and
local applications with calcipotriol and betamethasone propionate. Evolution was favorable with complete resolution
of lesions.
Acrodermatitis continua of Hallopeau is a localized form of pustular psoriasis.
Retinoids are the treatment of choice with the best long-term risk benefit.
Type 2 diabetes is associated comorbidity.
P1-021 PATTERN OF LUPUS NEPHRITIS IN THE MILITARY HOSPITAL:A HISTOLOGICAL AND
IMMUNOFLUORESCENCE STUDY
N. Al sheikh1
1Immunology,
Institute of Endemic diseases, khartoum, Sudan
Introduction: Lupus nephritis (LN) is the most common and serious manifestation of systemic lupus erythematousus
(SLE). There is a wide variation in the natural history of systemic lupus erythematosus (SLE) among different ethnic
and geographical groups. Studies in Arabs are few and those in North Africans and especially in the Sudanese
population donÕt exist. This study aims to characterize the pattern of Lupus nephritis among Sudanese patients.
Objectives: To demonstrate the clinical, laboratory features of the disease. The study was conducted at the Renal
Dialysis unit at The Military Hospital, Sudan. From April 2009 * May 2010.Atotal of 34 patients were studied. Results:
Of the 34 patients there were 33 females and one male between 10 -75 years with a median age of 24 years. The
histological pattern of lupus nephritis using World Health Organization classification, class I (n = 2, 5.9%); class II (n =
7, 20.6%); class III (n = 9, 26.5%); class IV (n = 8, 23.5%); class V (n = 3, 8.8%); class VI (n = 5, 14%). There was no
significant correlation between immunoglobulins & complement components performance and the WHO classes (p
value > 0.05 for all classes).The most common significant features at the time of biopsy were ≥ 3.5g of 24 hours urine
protein detected in (94.1%)of the patients and edema in (73.5%) of them. Conclusion: I conclude that focal
segmental glomerulonephritis (class III) is the most common histopathological pattern of Lupus nephritis. The
Sudanese pattern resembles that reported in some Arab countries such as Kuwait and KSA and differs from those
reported in African countries and other parts of the world.
Poster Session: Session 1: Autoimmunity, the heart and Artherosclerosis
P1-022 CLINICAL DETERMINANTS AND SIGNIFICANCE OF IMMUNOGENIC OXIDIZED LOW-DENSITY
LIPOPROTEIN/BETA2-GLYCOPROTEIN I (OXLDL/B2GPI) COMPLEXES IN ATHEROTHROMBOTIC
CARDIOVASCULAR DISEASES
L.R. Lopez1, K. Guyer2, E. Matsuura3, C.B. Rockman4, J.S. Berger5
1Clinical
Affairs, Corgenix Inc., Broomfield, USA
Indiana University South Bend., South Bend, USA
3Cell Chemistry, Okayama University Graduate School of Medicine Dentistry and Pharmaceutical Sciences,
Okayama, Japan
4Vascular Surgery, New York University School of Medicine, New York, USA
5Cardiology and Hematology, New York University School of Medicine, New York, USA
Background and Aims: OxLDL/β2GPI complexes have been implicated in the initiation and progression of
atherosclerosis. The clinical determinants and significance of oxLDL/β2GPI complexes were evaluated in arterial and
venous disease. Methods: 146 study subjects were recruited from a vascular surgery outpatient clinic: 61 had arterial
disease (21 carotid artery disease, 17 peripheral artery disease, and 23 abdominal aortic aneurysm), 32 had venous
disease, and 53 were healthy controls. Serum was obtained to measure oxLDL/β2GPI by ELISA. Results were
expressed in U/mL. Results: Mean oxLDL/β2GPI levels were significantly elevated in arterial (0.69±0.50 U/mL,
p=0.004) and venous groups (0.54±0.37 U/mL, p=0.025) compared to controls (0.39±0.33 U/mL). There was a
significant association of oxLDL/β2GPI with the following demographic and clinical variables: sex (female 0.26 vs
male 0.55 U/mL, p=0.005); age (r=0.299, p=0.002); hypertension (0.54 vs 0.27 U/mL, p=0.024) and family Hx of
previous clotting episode (0.95 vs 0.37 U/mL, p=0.035). Subjects with oxLDL/β2GPI values above the median (0.25
U/mL) were significantly more likely to have arterial disease (OR 2.5, 1.15-5.32, p=0.019) and venous disease (OR
2.8, 1.11-7.33, p=0.026). After excluding subject on statins, the odds for arterial disease was 4.5 (1.54-13.14,
p=0.004) and venous disease was 4.1 (1.38-11.99, p=0.008). Multivariate regression models with oxLDL/β2GPI as a
dependent variable indicated male gender (t=2.34, p=0.021), high cholesterol (t=2.58, p=0.011) and the CAD
phenotype (t=2.32, p=0.023) as significant predictors of oxLDL/β2GPI. Conclusions: OxLDL/β2GPI complexes are
elevated in patients with atherothrombotic vascular diseases. Male gender, hypercholesterolemia and the CAD
phenotype were significant independent predictors of oxLDL/β2GPI.
2Chemistry,
Poster Session: Session 1: Diagnostics: new chips, quick diagnostic tests
P1-024 AVAILABILITY OF THE MOST COMPLETE PANEL OF RECOMBINANT SM AND U1-SNRNP TARGETS
FOR IMPROVED SLE MULTIPARAMETER ASSAYS
I. Asen1
1Proteinchemistry,
DIARECT AG, Freiburg, Germany
Background and aims: Most frequently SmB and SmD polypeptides are targets of the anti-Sm specific autoimmune
response. Since SmB/B« and U1-snRNP specific proteins share a cross-reactive epitope motif, SmD is regarded the
most SLE-specific Sm antigen. A specific feature of SmD1, SmD3, and SmB/B' proteins is the symmetrical
dimethylation of arginine residues by specific protein arginine methyltransferases, which is important for regulating
the snRNP assembly. Symmetrically dimethylated SmD1 and SmD3 targets together with recombinant SmE/F/G
antigen may further improve SLE diagnostics when incorporated in novel microarrays.
Methods: Recombinant, full-length Sm and U1-snRNP proteins were expressed in and purified from baculovirusinfected insect cells. Clinically well-defined sera from patients with SLE were analyzed by various multiparameter
assay techniques such as dot blots, line assays and protein microarrays in standard microtiter plates.
Results: Symmetrical dimethylation of arginine residues in recombinant SmD1 and SmD3 is critical for their reactivity.
Preliminary clinical studies with well-characterized patient sera indicate applicability of the symmetrically dimethylated
targets as well as the recombinant SmE/F/G antigen in various assay formats.
Conclusion: Availability of the most complete panel of recombinant Sm targets represents an advancement for
established and novel assay formats. Together with recombinant U1-snRNP proteins they may significantly improve
existing multiparameter diagnostics of Systemic Lupus Erythematosus.
P1-025 COMPARATIVE ANALYSIS OF TWO LUMINEX-BASED SYSTEMS FOR DETECTION OF DOUBLESTRANDED DEOXYRIBONUCLEIC ACID ANTIBODIES
J. Lin1, C. Lee1, M. Cheng1
1Laboratory
Medicine, Changhua Christian Hospital, Changhua, Taiwan
Background: Testing for double-stranded deoxyribonucleic acid antibodies (anti-dsDNA) helps to predict and control
autoimmune diseases such as systemic lupus erythematosus flare-ups. The Luminex-based system is a convenient
platform that can simultaneously detect anti-nuclear antibodies (ANA) and anti-dsDNA. The aim of this study is to
compare two Luminex-based systems for anti-dsDNA testing in clinical sera.
Methods: One hundred and twenty-five clinical sera submitted for ANA or anti-dsDNA testing by the semi-automated
AtheNA Multi-Lite system were recruited. An aliquot of each sample was subsequently tested by the fully automated
BioPlex 2200 system. Anti-dsDNA results of both systems were compared for concordance after exclusion of
equivocal or indeterminate results. The recent fluorescent ANA (homogeneous) pattern or quantitative analysis of
anti-dsDNA by the enzyme-linked immunosorbent assay (ELISA) was regarded as reference for accuracy
assessment when discrepancy occurred.
Results: Twelve (9.6%) and fourteen (11.2%) sera were positive by AtheNA only and BioPlex only, respectively. Only
two sera were indeterminate by BioPlex and were excluded. The concordance rate (n=123) between systems was
95.1% (kappa coefficient, 0.74). Six discrepant sera, i.e., two positive by AtheNA and four positive by BioPlex were
detected. Accuracy evaluation was conducted in five of them having available refere information and revealed a false
negative rate of 80% and 20% by AtheNA and BioPlex, respectively.
Conclusions: Anti-dsDNA analyzed by AtheNA and BioPlex is comparable. AtheNA seems inferior in analytical
accuracy by expressing a higher false negative rate than BioPlex. Whether semi-automated analytical processes
contribute to such technical error needs to be investigated.
P1-026 DETECTION OF DENSE FINE SPECKLED PATTERN ON HEP-2 CELLS AND ANTI-DFS70 ANTIBODIES
BY ELISA-CLINICAL SIGNIFICANCE IN ASSOCIATION WITH SPECIFIC AUTOANTIBODIES
E.J. Oh1, H.Y. Lee1, Y. Kim1, Y.J. Park1, K. Han1
1Laboratory
Medicine, College of Medicine The Catholic University of Korea Seoul St.Mary's hospital, Seoul, Korea
Background and aims: While autoantibodies producing dense fine speckled pattern on HEp2 cells (HEp2-DFS)
have been reported to be more prevalent in healthy individuals than in systemic rheumatic diseases (SRD),
association with other specific antibodies or clinical significance of anti-DFS70 antibodies still remain unclear.
Methods: From ANA requested sequential 10,528 sera, HEp2-DFS(+) 181 (1.7%) sera were additionally tested for
anti-DFS70 antibodies by ELISA kit (ELISA-DFS70) and for specific autoantibodies (spANA; autoantibodies to Sm,
RNP, SSA, SSB, Scl-70, Jo-1, CENP, dsDNA, Ribosomal P, Chromatin). Fifty control sera which had HEp2-DFS()/spANA(+) were also tested for ELISA-DFS70.
Results: 181 HEp2-DFS(+) sera were from 64 SRD (22 SLE, 20 RA, 8 SjogrenÕs syndrome, 14 others) and 117
non-SRD patients. 109 (60.2%) sera were ELISA-DFS70(+) and 38 (21.0%) sera had spANAs. There was no spANA
in association with HEp2-DFS(+). ELISA-DFS70 was negative in all 50 control sera. Of 109 anti-DFS70 (+) sera, 26
(23.9%) sera also had spANA and 31 sera (28.4%) were from patients with SRD including 11 SLE patients. However,
of 83 HEp2-DFS(+)/ELISA-DFS70(+)/spANA(-), 18 (21.7%) sera were from SRD patients including only one SLE
patient. In 22 HEp2-DFS(+) SLE patients, 11 (50.0%) patients were ELISA-DFS70(+) and two SLE patients had
isolated ELISA-DFS70(+) without specific autoantibodies. SpANAs to dsDNA, Chromatin and SS-A60 were
significantly associated with HEp2-DFS(+)/ELISA-DFS70(-)/spANA(+).
Conclusion: In HEp2-DFS(+) sera, the additional ELISA-DFS70 and spANA tests could improve the efficiency of
diagnosing the SRD. Inclusion of ELISA-DFS70 in ANA test algorithm should be considered.
Poster Session: Session 1: Etiology and pathogenesis
P1-027 INVESTIGATING THE ROLE OF IL-20 IN THE PATHOGENESIS OF BEHÇET’S DISEASE
G. Keskin1, D. Keskin2, A. Inal3, B. Mavi2
1Immunology,
Ankara Üniversity Faculty of Medicine, Ankara, Turkey
DYB Training and Research Hospital, Ankara, Turkey
3Immunology, Gata, Ankara, Turkey
Background and aims: Beh*etÕs disease (BD) is a multisystem inflammatory disease. In BD, increased release of
proinflammatory cytokines may play a role in inflammatory stages of the disease. During the disease activity,
elevations in acute phase reactant levels and increased proinflammatory cytokines, such as IL-8, IL-6 and IL-22 have
been described. IL-20 is a proinflammatory cytokine and it has been reported to be involved in the development of
autoimmune diseases, such as lupus nephritis, psoriasis and RA. We aimed to search the relation among levels of
serum IL-20 with activity of BD.
2Immunology,
Methods: 64 patients with BD and 16 healthy controls were enrolled in this study. Thirty-eight patients were in active
stage (mean age; 34.9 ± 5.4 years, median disease duration 7 years) and 26 patients were in inactive stage (mean
age; 29.5 ± 6.9 years, median disease duration; 8 years). Serum IL-20 levels were determined by ELISA.
Results: The median serum IL-20 levels were 26,7 pg/ml in healthy controls, 232,6 pg/ml in active BD patients and
47,4 pg/ml in inactive BD patients. Serum IL-20 levels were significantly high in active BD patients compared with in
inactive BD patients (p<0.001) and healthy controls (p<0.001). In active patients with BD, there are statistically
significant correlation between serum IL-20 and serum CRP levels (r=0.674, p <0.001), ESR (r=0.602, p <0.001),
uveitis (r=0.557 p=0.01), oral ulcer (r=0.612 p=0.01) and arthritis (r=0.562, p=0.01).
Conclusions : IL-20 was associated with disease activity in BD. So, it may be involved in its pathogenesis.
P1-028 IMMUNOGENICITY OF PEROXYNITRITE-MODIFIED HSA AND ITS IMPLICATIONS IN CANCER
P. Ahmad1, K. Dixit1, Moinuddin1, R. Mahmood2, A. Ali1
1Biochemistry
F/o Medicine, Aligarh Muslim University, Aligarh, India
F/o Life Science, Aligarh Muslim University, Aligarh, India
Background: Peroxynitrite (ONOO?) is a potent oxidizing and nitrating agent. It can oxidize thiols and/or nitrate DNA,
proteins and lipids. 3-nitrotyrosine is an established biomarker of peroxynitrite damage to proteins. Being most
abundant plasma protein, human serum albumin (HSA) is quite vulnerable to oxidizing and nitrating agents in vivo
and hence, if go unchecked, could result in conformational alteration/structural modification, leading to changed
biological properties like reduced antioxidant function. Post-translational modifications of proteins by free radicals
may play a key role in increasing the onset of different cancers.
2Biochemistry
Materials and Methods: Commercially available HSA was modified with peroxynitrite and the structural
modifications were analyzed by different physicochemical and immunological studies. Cancer sera having high titre
anti-HSA autoantibodies were subjected to binding studies (enzyme immunoassay & gel retardation assay) with
native & peroxynitrite-modified HSA.
Results: The results of different physicochemical techniques showed alterations in secondary and tertiary structures
of HSA molecule and formation of 3-nitrotyrosine upon peroxynitrite exposure. Peroxynitrite-modified HSA was found
to be extremely potent antigen inducing high titre antibodies in experimental animal while native HSA resulted in
moderate antigenic response. We also observed preferential binding of cancer auto-antibodies with peroxynitritemodified HSA in comparison to native protein.
Conclusion: In conclusion, our study shows that peroxynitrite potentially change the secondary and tertiary structure
of HSA, which in turn leads to the generation of neo-epitopes in HSA molecule, thus making it highly immunogenic.
The binding studies of anti-HSA auto-antibodies of cancer patients suggest that alteration in the structure of HSA may
reduce the antioxidant property of the protein which in turn let different free radicals to easily damage the
biomolecules of the system and hence affects the onset/progression to cancer.
P1-029 MOLECULAR PATHOGENS ARE INVOLVED IN THE PATHOGENESIS OF AUTOIMMUNE DISEASES
AND CANCER
Z. Zou1
1Cancer
Pathology & Immunology, Hillside Laboratory, Beijing, China
Autoimmune dermatitis herpetiformis is infectious. The author transferred this disease to himself by touching the
diseased fluid of a patient and then his own mucous membranes. Ten months later, dermatitis herpetiformis occurred
on the skin of abdomen under his iron belt buckle. The disease appears on his arms under the sunlight in summer,
and on his thigh when copper and iron keys touched the skin in his pocket, indicating that disease risk factors trigger
and accelerate disease progression. It showed that an infectious agent is involved in the pathogenesis of dermatitis
herpetiformis. An increasing number of autoimmune diseases were transferred to animals and individuals. Cancer
was transferred by both intact tumor cells and cell-free filtrates. Niu MC, DeCarvalho S and colleagues transferred
cancer with tumor RNA in animals. Among disease molecules, only tumor RNA can transfer the disease to new host.
Nuclease degrades exogenous nucleic acids in blood plasma. Oxidants reduce nuclease activity and promote the
pathogenesis of autoimmune diseases and cancer. This raised a possibility that exogenous DNA and RNA molecules
named molecular pathogens are involved in pathogenesis of these diseases. These nucleic acids have low toxicity
and infectivity, thus, oxidants are required to reduce nuclease activity as prerequisite. Three key evidences support
the idea that a special category of exogenous nucleic acids is involved in the pathogenesis of these diseases. 1:
Increasing autoimmune diseases are transferred from the patients to other individuals and animals. 2: Oxidants
deactivate nuclease in disease development. 3: Cancer was transferred by tumor RNA.
Poster Session: Session 1: Experimental models
P1-030 THE EFFECTS OF CAFFEINE INGESTION BEFORE HALF BATH IN HOT WATER ON SERUM LEPTIN
LEVEL IN HUMANS.
T.W. Kim1, J.B. Lee2, Y.O. Shin1, Y.K. Min2, H.M. Yang2, H.S. Seo3
1Health
Care, Soonchunhyang University, Asan, Korea
College of Medicine Soonchunhyang University, Cheonan, Korea
3Exercise Prescription, Konyang University, Nonsan, Korea
We assessed the effects of ingesting caffeine before half bath in hot water on serum leptin and sweating responses,
which are both physiological responses associated with energy expenditure. The subjects were nine male university
students. This study used a within-subject, random, crossover design. Tests were performed twice at the same time
(2*5 p.m.) at a 1-week interval following 3 mgákg-1 caffeine ingestion or not. Half bath in hot water (42 ± 0.5¡C for 30
minutes) in a thermoneutral climate chamber (25 ± 0.5¡C, 60 ± 3% relative humidity, < 1 m/second air velocity).
Leptin, free fatty acids, waist size, mean whole body output volume and mean active sweat gland density increased
significantly after a single PHL session (P < 0.05). Leptin and all other parameters were significantly higher than
those after a single PHL session and caffeine ingestion before PHL (P < 0.05). A single PHL session not only
increased leptin secretion, lipolysis, and the sweating response but also increased energy expenditure via changes in
sympathetic nerve activity. The results suggest that ingesting caffeine before PHL is more energy efficient than that of
a single PHL session.
2Physiology,
P1-031 GALACTOSYLATED LIPOSOME AS A POTENT TARGETED CARRIER FOR ANTIGEN PRESENTING
CELLS
S.F. Lin1, P.L. Jiang2, H.J. Lin2, W.Y. Tsai1, M.Y. Chien1, S.W. Wang3, D.Z. Liu4
1Biomedical
Materials and Tissue Engineering, Taipei Medical University, Taipei, Taiwan
Institute of Biomedical Engineering College of Engineering College of Medicine, National Taiwan
University, Taipei, Taiwan
3School of Pharmacy College of Medicine, National Taiwan University, Taipei, Taiwan
4Center for General Education, Hsuan Chuang University, Hsinchu City, Taiwan
Background and aims:
2Graduate
Antigen presenting cells (APC), including dendritic cells (DC) and macrophages, are indispensable in the regulation
of the delicate balance between immunity and tolerance. A key role for DCs and macrophages has been suggested in
part by successful treatment for autoimmune diseases. In recent years some promising strategies has arisen over the
delivery of drugs, therapeutic genes, or immune-modulating molecules to APC for therapeutic purposes. Thus, our
aim was to develop a targeted carrier for more efficient delivery to APC.
Method and Results:
In the present study, we developed a galactosylated liposome carrier. The galactosyllipid was incorporated into a
liposome bilayer to form a galactosylated liposome carrier. We then evaluated the cellular uptake of galactosylated
liposomes by mouse bone marrow-derived dendritic cell and bone marrow-derived macrophage in vitro. Cells were
cultured with galactosylated liposome for 6 and 18 h. FACS analysis showed that galactosylated liposome carrier had
a higher uptake rate. Furthermore, mice were administrated with galactosylated liposomes via intravenous and
intranasal routes. The same results were observed in vivo.
Conclusion:
These data indicated that galactosylated liposomes are considered to be a predominant carrier for targeting antigen
presenting cells with drugs, therapeutic genes, or immune-modulating molecules to enable treatment of autoimmune
diseases.
Poster Session: Session 1: Gastrointestinal autoimmunity
P1-032 PANCREATIC MAJOR ZYMOGEN MEMBRANE GLYCOPROTEIN2 ( MZGP2) IGG AND IGA
AUTOANTIBODIES ARE SPECIFIC MARKERS FOR THE DETECTION OF CROHN'S DISEASE
G.L. Norman1, Z. Shums1, P. Pavlidis2, A. Forbes3, D.P. Bogdanos2
1R
& D, INOVA Diagnostics Inc., San Diego, USA
of Medicine, Kings College Hospital, London, United Kingdom
3Gastroenterology and Clinical Nutrition, University College Hospital, London, United Kingdom
Introduction: ASCA (anti-Saccharomyces cerevisiae antibody) and ANCA (antineutrophil cytoplasmic antibody)
assays are used to aid the non-invasive differentiation of Crohn's Disease (CD) and ulcerative colitis (UC). ASCA has
sensitivity of 50-65% for CD and ANCA a sensitivity of 45-70% for UC. Up to 30% of CD patients have antibodies to
structures in the exocrine pancreas. These antibodies are mainly directed against the major zymogen granule
membrane glycoprotein2 (MZGP2).
2School
Aim: Evaluate the performance of QUANTA Lite® MZGP2 IgG and IgA ELISA assays (RUO, INOVA Diagnostics) in
IBD patients and controls.
Methods: Sera from a well-characterized IBD cohort from University College Hospital, London, (125 CD and 123 UC)
were tested on Quanta Lite® MZGP2 IgG and IgA ELISAs. Controls included 103 healthy and 112 disease controls.
Results: MZGP2 IgG was detected in 33/125 (26%) and MZGP2 IgA in 17/125 (14%) of the CD patients. MZGP2
IgG and IgA were positive in 4/123 (3.2%) and 5/123 (4%) of the UC patients respectively. MZGP2 IgG and IgA were
detected in 7/215 (3.2%) and 6/215 (2.8%) of the healthy and disease controls, respectively. Adding MZGP2 IgG/IgA
to ASCA testing increased overall sensitivity for Crohn's disease to 72%, uniquely detecting 5/125 (4%) CD patients.
Conclusion: The QUANTA Lite® MZGP2 IgG and IgA ELISAs contributed to the differential diagnosis of CD in IBD
with a sensitivity of 26% including the unique identification of 4/82 (4.9%) CD patients. Adding MZGP2 to the
traditional panel of serological markers may help to accurately detect more CD patients using non-invasive methods.
P1-033 CHRONIC GASTRITIS AS THE GASTROINTESTINAL MANIFESTATION IN SYSTEMIC VASCULITIS
A. Kurniawan1, N. Lugito1, M. Tjiang1, I. Wijaya1, T. Yanto1, R. Setiawan1
1Internal
medicine, University of Pelita Harapan, Jakarta, Indonesia
Introduction
Vasculitis can cause local or diffuse pathologic changes in the gastrointestinal tract, resulting in nonspesific paralytic
ileus, mesenteric ischemia, submucosal edema and hemorrhage, or bowel perforation or stricture. Sytemic vasculitis
is known to affect the gastrointestinal tract but the nature of the complication is poorly charaterized.
Case report
We reported a 48 year old man came with multiple ulcer in mouth and right leg. He also felt fever, erythema in his left
eye, epigastric pain, black ter like feces, and decrease his body weight. He did not complaint about edem in his leg or
others place. From physical examination revealed redness in his left eye with loss his sight, muliple ulcer in buccal,
epigastric pain, ulcer in his leg with 3-4 cm in diameter with no pus. From the laboratoy results showed normocytic
normochromic anemia, leukocytosis with netrophilia, increased D-dimer, hypoalbuminemia, ANA positive with
speckled pattern; complement C3/C4 was 76,4/140. Skin biopsy of leg ulcer showed vasculitis. Gastroscopy result
showed erosive gastritis and colonoscopy result showed multiple ulcer in colon. Result of biopsy of gastric showed
the presence of vasculitis which patohological anatomic result revealed signs of erythrocyte extravacation. He was
given methyl prednisolon at immunosuppresant dose, oral anticoagulant with prophylactic dose, proton pump inhibitor
and acyclovir. The ulcers were resolved after one month follow up.
Key Words: Systemic vasculitis, multiple ulcer, non active chronic gastritis
P1-034 PLUMMER VINSON SYNDROME-A CASE REPORT
D. Madi1, S. Yadalpati1
1Medicine,
MANIPAL UNIVERSITY, mangalore, India
Plummer Vinson syndrome represents the classical association of post cricoid dysphagia, upper oesophageal webs
and iron deficiency anemia . Plummer Vinson Syndrome is a rare cause of dysphagia. Proposed etiologic factors are
iron, nutritional deficiencies and autoimmunity.
A 46 year old female presented to our hospital, with complaints of easy fatiguability and dysphagia of one year
duration. Clinical examination revealed pallor and platynychia.
Her investigations showed haemoglobin of 5.6gm%(11-15 gm%), Serum Ferritin - 1.55 ng/ml and peripheral smear microcytic, hypochromic anemia. Barium swallow (Figure 1) and UGI scopy was done which showed a mucosal web
causing luminal compromise.Endoscopic dilation of the web was done. She was transfused with 2 units of packed
cells.Iron supplements were given.
A diagnosis of Plummer Vinson syndrome should be considered in any person presenting with iron deficiency anemia
and dysphagia because prognosis of Plummer-Vinson syndrome is excellent and it is a reversible cause of
dysphagia.
P1-035 CALPROTECTIN AND ANCA IN NEUTROPHIL-MEDIATED IBD
I. Lochman1, A. Lochmanová2, J. Martinek3, H. Tomaskova4, I. Bártková5, P. Svoboda6, V. Smajstrla7
1Immunology,
Spadia Lab Ltd, Ostrava, Czech Republic
of Biomedical Sciences, Faculty of Medicine University of Ostrava, Ostrava, Czech Republic
3Immunology and Allergy, Institute of Public Health, Ostrava, Czech Republic
4Department of epidemiology and Public Health, Faculty of Medicine University of Ostrava, Ostrava, Czech Republic
55Department of Pediatrics, Municipal Hospital, Ostrava
66Department of Clinical Studies, Faculty of Medicine, University of Ostrava
77Bormed, Ltd., Ostrava, Czech Republic
Determination of faecal calprotectin and ANCA are used in the diagnosis and laboratory monitoring of inflammatory
bowel diseases - IBD (Crohn's disease, ulcerative colitis and neoplasms). Both analytes are associated with an
inflammatory process which involves neutrophils, but the information obtained from these tests is not completely
identical.
2Department
We analyzed the results of faecal calprotectin using Quantum blue (Buehlmann) and ELISA (R-Biopharm) kits and
ANCA determination using IFA CIBD-Profile 1 kits (Euroimmun) containing chips with ethanol-fixed granulocytes and
IgG conjugate in our labs during 2011 and 2012. Overall, 1088 samples from 823 patients have been analyzed for
calprotectin and ANCA and 4723 samples only for ANCA using IFA CIBD-Profile 1 kits.
Detailed analysis of results confirmed that the levels of both analytes correlate with the intensity of neutrophilmediated inflammation. We demonstrate it on several case reports. The vast majority of total ANCA was designated
as A-ANCA, possibly GS-ANA after typing, but it was difficult or impossible to distinguish these antibodies in the
presence of other antinuclear antibodies in some cases.
However targets against which these antibodies are formed, are mostly NETs epitopes (neutrophil extracellular traps)
that is difficult, if not impossible, to define generally. Eventhough IFA on ethanol-fixed granulocytes is generally
recommended for ANCA screening, it has not been uniquely accepted legal diagnostic scheme for ANCA screening
and their subsequent typing. Especially for diseases other than ANCA associated vasculitis (AAV) can be obtained
frequently confused results between IFA and other immunoassays that use defined neutrophil cytoplasmic
substrates.
P1-036 THE TETRASPANIN CD151 AS A NOVEL REGULATOR OF T CELL MOTILITY IN HEALTH AND
INFLAMMATORY BOWEL DISEASE
E. Toister-Zelman1, E. Bakos1, S. Cohen1, E.H.U.D. Zigmond1, V. Grabovsky1, A.D.I. Sagiv1, G.I.L.I. Hart1, N.
Kaushansky1, A.V.I. Ben-Nun1, A. Sonnenberg1, R. Alon1, I.D.I.T. Shachar1
1Department
of Immunology, The Weizmann Institute of Science, Rehovot, Israel
The continuous recirculation of mature lymphocytes and their entry into the peripheral
lymph nodes (LNs) are crucial for the development of an effective immune response.
Picomolar (pM) levels of CCL2 were previously shown to inhibit T cell migration to LNs
resulting in reduced inflammation. However, the molecular mechanisms regulated by low
dose CCL2 remained unknown. In this study, we demonstrate that the tetraspanin CD151 is a target
of CCL2 in T cells. pM levels of CCL2 downregulated CD151 expression and dissociated
CD151/VLA-4 and CD151/LFA-1 complexes in T lymphocytes, resulting in their attenuated
migration. Ablation of CD151 expressed on immune cells reduced inflammation in a DSS
induced colitis model of CD151-/- chimeric mice. Moreover, blocking of CD151, using CD151
truncated peptide fragment, reduced cell migration in vitro and in vivo, and inhibited the
development of DSS-induced colitis. Thus, CD151 is a novel orchestrator of T cell motility,
and its targeting by inhibitory peptides results in beneficial suppression of inflammation.
Poster Session: Session 1: Genetics and autoimmunity
P1-037 ASSOCIATION OF INTERLEUKIN-10 -1082A/G POLYMORPHISM IN BULGARIAN PATIENTS WITH AS
AND RA
M. Ivanova1, I. Manolova2, A. Avramova3, N. Stoilov1, R. Stoilov1, R. Rashkov1, S. Stanilova3
1Clinic
of Rheumatology, University Hospital "Sv. Iv. Rilski", Sofia, Bulgaria
of Health Care, Medical Faculty Trakia University, Stara Zagora, Bulgaria
3Department of Molecular Biology Immunology and Medical Genetics, Medical Faculty Trakia University, Stara
Zagora, Bulgaria
Background: IL-10 is an important regulatory cytokine with a significant role in autoimmune diseases. The promoter
polymorphism in IL-10 gene (IL10), located at -1082 A/G (rs1800896) has an impact on IL-10 production. Studies on
the relationship of IL-10 rs1800896 polymorphism with RA and AS are inconclusive.
2Department
Objective: To investigate whether IL10 rs1800896 enables development of AS and RA in Bulgarian population.
Patients and methods: Genotyping for -1082A/G polymorphism in IL10 was performed by allele specific PCR. 58
patients with AS and 56 with RA were compared with 106 and 88 matched healthy subjects, respectively.
Results: No association between the rs1800896 polymorphism and RA risk was established under the additive
model (allele A vs allele G; OR=0.99), the codominant model (AA vs. GG; OR=1.061), the dominant model (AA+AG
vs. GG; OR=1.13) and the recessive model (AA vs. AG+GG; OR=1.08). An association between AS and rs1800896
polymorphism in the same genetic models (additive: OR=1.374, 95%CI=0.83ü2.27, p=0.19; codominant: OR=1.607,
95%CI=0.519ü5.095, p=0.51; dominant: OR=1.201, 95%CI =0.42ü3.5, p=0.89; recessive: OR=0.59,
95%CI=0.29ü1.19, p=0.113) was found. These results suppose that AA genotypes could be predisposing, whether
GG genotypes might be protective for susceptibility to AS. Moreover, significant associations were observed in AS for
age at disease onset (age at onset <29; OR=2.769, 95%CI =0.836ü9.382, p=0.06) and functional outcome (BASFI
score>4; OR=3.956, 95%CI =1.137ü14.245, p=0.014).
Conclusion: Promoter polymorphism -1082A/G in IL-10 gene has no significant effect on RA development, but could
play a role in AS, mostly determining age of disease onset and disease severity in Bulgarian patients.
P1-038 THE RELATIONSHIP BETWEEN SYSTEMIC LUPUS ERYTHEMATOSUS AND CLEC16A, A PUTATIVE
NOVEL C-TYPE LECTIN-LIKE PROTEIN
R. Tam1, S. Yan1, W.C.S. Lau1, S.F.V. Chan1
1Medicine,
The University of Hong Kong, Hong Kong, Hong Kong China
Several independent genome wide association studies have shown associations of single nucleotide polymorphisms
within the C-type lectin domain family 16 member A gene, CLEC16A, with various autoimmune diseases, including
systemic lupus erythematosus (SLE). However, the expression and function of this novel C-type lectin domaincontaining protein is still largely unknown. This study aimed to relate its expression level in peripheral blood
mononuclear cells (PBMCs) with SLE disease activity. Two expressed isoforms, isoforms 1 and 2, of CLEC16A were
cloned and sequenced from PBMCs from healthy Chinese. Surprisingly, both sequences of isoforms 1 and 2 bore
differences when compared to the reference sequences derived from Caucasians. In isoform 1, exon 5 was
consistently absent in Chinese PBMCs. However, in isoform 2, the stretch of 48-bp in-frame deletion within exon 11
in the reference sequence was found present in Chinese PBMCs. The mRNA expression levels of both isoforms were
found significantly lower in SLE patients than in normal individuals. Yet, their expression levels did not correlate with
disease activity as measured by SLE disease activity index. The molecular and cellular functions of CLEC16A are
currently under investigation, which should shed light on the importance of this protein in SLE pathogenesis.
P1-039 ASSOCIATION OF +3179 A/G IGF-1R POLYMORPHISM WITH RHEUMATIC DISEASES (RA AND AS)
S. Stanilova1, M. Ivanova2, I. Karakolev1, R. Stoilov2, R. Rashkov2, I. Manolova3
1Department
of Molecular Biology Immunology and Medical Genetics, Medical Faculty Trakia University, Stara
Zagora, Bulgaria
2Clinic of Rheumatology, University Hospital "Sv. Iv. Rilski", Sofia, Bulgaria
3Department of Health Care, Medical Faculty Trakia University, Stara Zagora, Bulgaria
Background: The insulin-like growth factor (IGF) system plays a prominent role in the regulation of immunity and
inflammation. Inappropriate balance of the IGF-1 signalling has been reported in autoimmune disorders. A few
previous studies have investigated the modulation of the IGF signalling pathway in the rheumatic diseases, but their
findings are controversial. Recently, single nucleotide polymorphism (SNP) located in exon 16 (+3179G/A;
rs2229765) of IGF-1R gene was found associated with serum levels of IGF-I and human longevity.
Aim: To compare +3179G/A IGF-1R genotype distribution in rheumatoid arthritis (RA) and ankylosing spondylitis
(AS).
Patients and methods: Genotyping for +3179G/A polymorphism was performed by restriction fragment length
polymorphisms (RFLP)-PCR assay in 70 patients with RA, 55 with AS and corresponding age and sex matched
healthy donors.
Results: We found an opposite effect of the investigated IGF-1R polymorphism (+3179G/A; rs2229765) on disease
susceptibility to RA and AS. Significant differences in allele and genotype frequencies between RA and healthy
controls were observed: higher frequency of homozygous genotype AA (22.9% vs14.1%; OR= 2.33, p=0.034) and
allele *ÒAÓ (47.9% vs37.5%; OR= 1.53, p=0.032) in cases versus controls. Although the genotype frequencies in AS
in comparison with healthy controls were not significantly differing, the lowest frequency of AA genotype was
observed in AS (10.9% vs 20,9%; OR= 0,47; p=0.142).
Conclusions: Allele A and genotype AA of +3179G/A IGF-1R polymorphism are associated with RA genetic
predisposition, but it seems as a protective factor for AS. Our data confirm the involvement of the IGF-1
signal pathway in RA.
P1-040 SUSCEPTIBILITY OF TNFAIP3 GENE POLYMORPHISMS TO RHEUMATOID ARTHRITIS AND
SYSTEMIC LUPUS ERYTHEMATOSUS IN KOREAN POPULATION
S.K. Kim1, J.Y. Choe1, S.H. Park1, H. Lee1, K.Y. Park2, J.H. Kim2
1Rheumatology,
Daegu Catholic university Hospital, Daegu, Korea
and Autoimmunity Research Center, Daegu Catholic university Hospital, Daegu, Korea
Objective: We investigated polymorphisms of TNFAIP3 gene to genetic susceptibility of systemic lupus
erythematosus (SLE) and rheumatoid arthritis (RA) in Korean population.
2Arthritis
Methods: Study participants included 422 patients with RA, 133 patients with SLE, and 422 healthy controls in this
case-control study. Genotyping for rs5029941(C>T), rs2230926(T>G), rs5029930(C>A), rs5029937(G>T),
rs5029939(G>C) in TNFAIP3 gene polymorphisms were performed. Status of RA-related autoantibodies including
rheumatoid factor and anti-cyclic citrullinated peptide (anti-CCP) in RA and presentations of arthritis or nephritis in
SLE were assessed.
Results: Significantly different frequencies of minor alleles in two TNFAIP3 polymorphisms in SLE patients were
noted, in comparison to healthy controls (OR 2.13, 95% CI 1.25-3.65, p = 0.02 for rs5029937; OR 2.17, 95% CI 1.273.72, p = 0.01 for rs5029939). In addition, SLE patients showed different frequencies of haplotypes compared to
healthy controls (p < 0.001). However, lack association between RA susceptibility and TNFAIP3 polymorphisms was
identified (p = 0.28). Interaction between RA-related autoantibody status and TNFAIP3 polymorphisms was not
associated with RA susceptibility. Interestingly, appearance of arthritis in SLE seemed to be marginally related with
TNFAIP3 polymorphisms (p = 0.04).
Conclusion: Our study suggests that TNFAIP3 gene polymorphism may be related with differential susceptibility to
SLE and RA in Korean population. It implicates that TNFAIP3 gene polymorphism to RA susceptibility might be
dependent on individual ethnics.
P1-041 AUTOSOMAL DOMINANT HYPERIMMUNOGLOBULIN E SYNDROME IN A 9 YEAR OLD BOY
A. Lubis1, R. Akbar1, Z. Hikmah1, A. Endaryanto1, A. Harsono1
1Child
health, Airlangga University, Surabaya, Indonesia
Background: The autosomal dominant hyperimmunoglobulin E syndrome is a rare primary immunodeficiency that
difficult to diagnose, because this disease has multisystem disorder.
Aims: To present a case of autosomal dominant hyperimmunoglobulin E syndrome in a 9 year old boy
Case report: A 9 year old boy with immunologic manifestation: skin cold abscesses, eczematous, excoriated,
lichenification, reccurent cutaneous infection, bilateral benign chronic suppurative middle otitis, bronchopneumonia,
and septicemia with blood culture result was Staphylococcus Aureus and skin culture result were Staphylococcus
Aureus and Pseudomonas Aeruginosa. Total immunoglobulin E were elevated (18,000 IU/ml and 50,000 IU/ml) and
eosinophils count 3,000 cells/ml. Non immunologic manifestation: coarse facial features, prominent forehead and
chin, broad nose, retained primary teeth, delayed bone age and also cardiac involvement. Based on United States
national health institute scoring system with clinical and laboratory test for individuals in kindreds with
hyperimmunoglobulin E syndrome the score is 53 and suggestive of autosomal dominant hyperimmunoglobulin E
syndrome. The treatment were supportive, antibiotics based on culture result and intravenous gammaglobulin were
given
Conclucions: Autosomal dominant hyperimmunoglobulin syndrome is a rare primary immunodeficiency that has
multisystem disorder with immunologic and non immunologic manifestation. Clinicians must depend on recognizing
this multisystem disorder and clinical features which develop over years, thereby making the diagnosis of
hyperimmunoglobulin E syndrome is difficult.
Keywords: Autosomal dominant hyperimmunoglobulin E syndrome, multisystem disorder
P1-042 SHARED AND DISTINCT GENETIC RISK FACTORS OF AUTOIMMUNE DISEASES
D. Chang1, A. Keinan1
1Biological
Statistics and Computational Biology, Cornell University, Ithaca, USA
The shared pathogenesis between autoimmune diseases has been explored through comorbidity studies. Genomewide association studies (GWAS), which compare genetic data between cases and controls, provide support of many
genetic risk factors shared between diseases. Studies have specifically explored this shared pathogenesis via GWAS
summary statistics alone; such as testing the correlation of association signals between pairs of autoimmune
diseases across many SNPs. Here we introduce a new method that rigorously explores the shared genetic risk
between autoimmune diseases and between them and diseases of other classes. Our method applies principal
component analysis (PCA) to a matrix of gene-level significance scores across many GWAS datasets, while
correcting for confounders such as genotyping arrays and sample size. We applied our method to 30 datasets, of
which 14 are autoimmune diseases. Other disease categories include psychiatric, neurological and cancers. We
report several findings: 1) different studies of the same diseases generally lie closer together. 2) Inflammatory bowel
diseases cluster together away from other autoimmune diseases. 3) Some psychiatric disorders cluster with
autoimmune diseases. 4) Genes that underlie the observed structure are significantly enriched for immune related
pathways such as IL1-mediated signaling. Prompted by our findings, we carried out a GWAS with combined cases of
bipolar and type-1 diabetes (T1D). We found an association near the gene PTPRM, which presents a new
association to T1D. In summary, our method offers novel findings regarding the shared pathogenetics of autoimmune
diseases.
P1-043 A POLYMORPHISM (RS6457617) BETWEEN HLA-DQB1 AND HLA-DQA2 WAS ASSOCIATED WITH
RHEUMATOID ARTHRITIS: A REPLICATION STUDY IN A CHINESE HAN POPULATION
H. Li1, Y.H. Hu2, X.M. Su1, Y.J. Zhu1, T. Zhang1, Y. Liu3, Y.T. Wang3, T. Yang3, M.H. Li3, Q.L. Luo1, Y. Cheng1, Q.
Zou3
1Department
of oncology, Chengdu Military General Hospital, Chengdu, China
of traditional Chinese medicine, Chengdu Military General Hospital, Chengdu, China
3Department of immunology, Chengdu Medical College, Chengdu, China
Background and aims: Previous genome-wide association study by WTCCC identified rheumatoid arthritis (RA)
susceptibility loci in British. Here, we recruited 1894 subjects to investigate whether the three SNPs with strong RA
association signal in British was associated with RA in Han Chinese. Methods: We analyzed the three SNPs in Han
Chinese population based on the HCB data of HapMap. The rs6679677 does not exist in Han Chinese. Other two
SNPs (rs6457617 and rs11761231) were performed case-control study in the test cohort of 380 subjects firstly and
the positive result was further replicated in another validation cohort of 1514 subjects. Results: In the test cohort, only
rs6457617 was significantly associated with RA. The individuals bearing the homozygous genotype CC had 0.39-fold
risk than these bearing the wild-type genotype TT (P = 0.004, OR 0.39, [95%CI 0.21-0.74]). And the protective effect
of allele C was confirmed in combined analysis with test and validation cohort(Pgenotye CC/TT = 9.4*10-12, OR 0.35, [95%
CI 0.26-0.47], Table). Conculsions: The rs6457617, which locates between the HLA-DQB1 and HLA-DQA2 encoding
β chain of MHC-II protein, was also associated with RA in Han Chinese.
Table. Validation of rs6457617 in test cohort, validation cohort and combined analysis
2Department
rs6457617
Test cohort
validation cohort
combined samples
subjects no.(%)
codominant genetic model
controls
cases
OR (95% CI)
P
TT
68 (37.0)
97 (52.4)
1 (ref)
-
CT
82 (44.6)
69 (37.3)
0.59 (0.38-0.92)
0.02
CC
34 (18.5)
19 (10.3)
0.39 (0.21-0.74)
0.004
TT
263 (35.7)
353 (48.2)
1 (ref)
-
CT
340 (46.1)
319 (43.5)
0.70 (0.56-0.87)
1.0×10-3
CC
134 (18.2)
61 (8.3)
0.34 (0.24-0.48)
5.9×10-10
TT
331 (35.9)
450 (49.0)
1 (ref)
-
CT
422 (45.8)
388 (42.3)
0.68 (0.56-0.82)
1.1×10-4
CC
168 (18.3)
80 (8.7)
0.35 (0.26-0.47)
9.4×10-12
geno- type
Data shown in n (%). OR = odds ratio; 95% CI = 95% confidence interval; genotype analyses in codominant genetic
model for association were used by Logistic Regression with SPSS software (version 13.0, SPSS Inc.).
P1-044 THE TRAF1/C5 (RS3761847) AND THE RISK OF RHEUMATOID ARTHRITIS: A CASE-CONTROL STUDY
INDIVIDUALLY MATHCED ON SEX
H. Li1, Y. Hu2, T. Zhang1, Y. Liu3, X. Su1, Y. Zhu1, Y. Wang3, T. Yang3, M. Li3, Q. Luo1, Y. Cheng1, Q. Zou3
1Department
of Oncology, Chengdu Military General Hospital, Chengdu, China
of traditional Chinese medicine, Chengdu Military General Hospital, Chengdu, China
3Department of Immunology, Chengdu Medical College, Chengdu, China
Background and aims: A SNP (rs3761847) in TRAF1-C5 locus on chromosome 9 was found to be an increased risk
for anti-CCP-positive rheumatoid arthritis (RA) by the genome-wide study in the North American Rheumatoid Arthritis
Consortium (NARAC) and the Swedish Epidemiological Investigation of Rheumatoid Arthritis (EIRA)[1]. Here, we
investigated whether the susceptibility loci in Caucasian was associated with RA in Han Chinese population.
Methods: 190 RA patients and 190 healthy controls among Chongqing city residents were evaluated in this study.
Since RA was the sole disease for which the sex-differentiated analysis generated a strong signal due to different
genetic effects in males and females, a single healthy control was matched with each case on the basis of sex, age
and ethnicity to exclude age and sex bias. rs3761847 was genotyped in 380 subjects by Genotype-specific PCR.
Results: Allele and genotype frequencies for the rs3761847 were in HWE in both the patient and the control
populations. The minor G allele frequencies for RA case group did not differ significantly from those in the control
group (43.7% versus 56.3%, P = 0.31; resulting OR = 0.86; 95% CI [0.65, 1.15], shown in the Table). The difference
in the frequency of the GG genotype between patients and controls failed to reach statistical significance (15.8%
versus 21.3%, P = 0.24; OR = 0.69; 95% CI [0.38, 1.28]). Conclusions: No evidence of the association between
rs3761847 and RA was observed in our individually matched case-control study.
2Department
Acknowledgments: This work was supported by grants from the National Natural Science Foundation of China (No.
81072455), The Basic Research Program of Sichuan Province of China (No. 2012JY0031), Research Fund of
Chengdu Military General Hospital (No.2011YG-B02 and No. 2013YG-A004), and Key Project of Chengdu Military
Region during the 12th five-year plan period (No.A12003).
Table: association anlysis of rs3761847
P
allele/genotypes controls no.(%) cases no. (%) OR
95%CI
A
198 (52.7)
213 (56.3)
1(reference) G
178 (47.3)
165 (43.7)
0.86
0.65-1.15 0.31
AA
50 (26.6)
54 (28.6)
1(reference) AG
98 (52.1)
105 (55.6)
0.99
0.62-1.59 0.97
GG
40 (21.3)
30 (15.8)
0.69
0.38-1.28 0.24
Data shown in n (%).OR = odds ratio; 95% CI = 95% confidence interval; Allele analysis for association were used by
Pearson Chi-Square test and genotype analyses for association were used by Logistic Regression with SPSS
software (version 13.0, SPSS Inc.).
1. Plenge RM, Seielstad M, Padyukov L, Lee AT, Remmers EF, Ding B, Liew A, Khalili H, Chandrasekaran A, Davies
LR et al: TRAF1-C5 as a risk locus for rheumatoid arthritis--a genomewide study. N Engl J Med 2007, 357(12):11991209.
P1-045 ASSOCIATION OF IL-1F7 GENES WITH SUSCEPTIBILITY TO RHEUMATOID ARTHRITIS IN HAN
CHINESE POPULATION
X. Zhang1, C. Li1, R. Mu1, Y. Zuo1, Y. Du1, X. Wu1, X. Lu2, R. Li1, J. He1, X. Liu1, F. Yin3, J. Guo1, Z. Li1
1The
department of Rheumatology and Immunology, People's Hospital Peking University health science center,
Beijing, China
2Department of Rheumatology, Xuanwu Hospital, Beijing, China
3The department of Rheumatology and Immunology, The First Affillated Hospital Baotou Medical College, Baotou,
China
Background IL-37 is a natural inhibitor of inflammatory and immune responses and IL1F7 polymorphisms have been
found to correlated with inflammatory diseases. This study was undertaken to investigate the role for genetic variants
in IL-37 in RA susceptibility.
Methods Genotypes for five selected single-nucleotide polymorphisms (SNPs) in IL1F7 genes (rs 2723186,
rs3811046, rs4241122, rs4364030, rs4392270) were determined by TaqMan R Allelic Discrimination in Han Chinese
population (727 RA cases and 610 healthy controls). Association analyses were performed on the whole data set and
on RA subsets based on the status of anti-cyclic citrullinated peptide antibody. Association statistics were calculated
by age and sex adjusted logistic regression.
Results No significant differences of distribution of alleles and genotypes were observed between case and control
groups (P>0.05). The haplotype analysis showed that IL-1F7 genes were not significantly associated with
susceptibility to RA.
Conclusions No significant correlation between RA susceptibility among the Han Chinese population and IL-1F7
genes is observed.
Table 1 Analysis of 5 SNPs in the IL-1F7 region in a Chinese rheumatoid arthritis population.
SNP
Allele
Cases
Minor/MajorN
rs2723186
rs3811046
rs4241122
rs4364030
rs4392270
A/G
G/T
G/A
G/C
A/G
687
690
340
346
349
2/1
197/1177
278/1102
136/680
341/692
91/607
11
12
22
n (%)
n (%)
n (%)
503
171
13
(73.2)
(24.9)
(1.9)
440
222
28
(63.8)
(32.2)
(4.0)
215
114
11
(63.2)
(33.5)
(3.2)
94
163
89
(27.2)
(47.1)
(25.7)
262
83
4
(75.1)
(23.8)
(1.1)
1, major (common) allele; 2, minor (rare) allele.
* After
Controls
Bonferronl correction, P value is not considered significant.
MAF
0.14
0.20
0.20
0.49
0.13
N
581
603
402
430
407
2/1
179/983
244/1206
202/804
436/860
110/814
11
12
n (%)
n (%)
408
167
(70.3)
(28.7)
407
178
(67.5)
(29.5)
211
180
(52.8)
(44.8)
101
222
(23.5)
(51.6)
302
100
(74.2)
(24.6)
Poster Session: Session 1: Hormones: vitamin D, prolactin
P1-046 EFFECT OF VITAMIN D ON THE NUMBER AND FUNCTION OF THELPER 17 CELLS (CD4+IL-17A+) IN
SLE PATIENTS WITH HIPOVITAMIN D
C.S. Wahono1, D. Soelistyoningsih2, K. Handono3, H. Kalim1, A.T. Endarti4
1Internal
Medicine Faculty of Medicine, Brawijaya University, Malang, Indonesia
of Biomedical Science Faculty of Medicine, Brawijaya University, Malang, Indonesia
3Clinical Pathology Faculty of Medicine, Brawijaya University, Malang, Indonesia
4Parasitology Faculty of Medicine, Brawijaya University, Malang, Indonesia
Objective: To determine the effect of vitamin D [1,25 (OH) 2D3] in the number and function of Th17 cells (CD4+IL17A+) in SLE patients with hipovitamin D.
2Laboratory
Methods: CD4+ T lymphocytes from PBMCs of 5 SLE patients with hipovitamin D were cultured using RPMI 1640,
PMA, ionomycin, Golgiplug, 10% heat-inactivated FCS, penicillin (100 IU/mL), and streptomycin (100 IU/mL). After
stimulation with 10 ng/mL IL-6, 5 ng/mL TGF-β1, 10 µg/mL anti-IFN-γ, and 10 µg/mL anti-IL-4, and treated with
various doses of 1,25 (OH) 2D3 ( P0: 0 M, P1:10-9M, P2: 10* 8 M, P3: 10-7 M), we analysed the the number and
function of Th17 cells. The number (percentage) of Th17 cells was measured by flowcytometry, while Th17 cell
function is measured by the levels of IL-17A expression using ELISA method.
Results: Compared with P0 (17:07 ± 2.99%), the mean percentage of Th17 cells in P1 (8:39 ± 3:29%; p = 0.043) and
P2 (7:17 ± 3.81%; p = 0.038) were significantly lower. Similarly, the levels of IL-17A, in P1 (25.90 ± 11.90 pg/ml; p =
0.024) and P2 (15.75 ± 1.22 pg/ml; p = 0.047) were significantly lower than P0 (59.18 ± 26.95 pg/ml).
Conclusion: Administration of vitamin D [1,25 (OH) 2D3] can reduce the number (percentage) of Th17 cells and IL17A levels in CD4+ Tcells culture of SLE patients with hipovitamin D.
Keywords: vitamin D, SLE, Th17, IL-17A
Poster Session: Session 1: Infection and autoimmunity
P1-047 HERPES ZOSTER INFECTION AMONG FILIPINOS WITH SYSTEMIC LUPUS ERYTHEMATOSUS AT A
SINGLE TERTIARY CARE CENTER
L.D. Zamora1, S.N. Leynes1, S.V. Navarra1
1Medicine
Section of Rheumatology, University of Santo Tomas Hospital, Manila, Philippines
BACKGROUND AND OBJECTIVE: Herpes-zoster (HZ) has higher occurrence in systemic lupus erythematosus
(SLE) compared to the general population. We aimed to identify clinical factors associated with HZ infections among
Filipino SLE patients.
METHODS: We reviewed the medical records ofSLE patients seen at the University of Santo Tomas (UST) Hospital
from 2008-2012 who were diagnosed with HZ, and described the following: site and dermatomal distribution of HZ,
SLE disease characteristics, immunosuppressive use, response to anti-viral therapy and HZ recurrences.
RESULTS: The medical records of 320 SLE patients were reviewed for the occurrence of HZ. Of these, 52 (16.3%)
developed HZ, with 4 patients having more than one episode of HZ. There were 48 (92%) females, the mean age
was 34.7 years (range 10-65), and mean SLE disease duration at HZ infection was 50.4 months (range 2-204). The
HZ lesions were all localized involving up to 3 dermatomes; none had disseminated disease. All cases were
responsive to antiviral therapy. Average prednisone dose was 17.3 mg/day (range 0-60), with 80 % of patients also
on hydroxychloroquine within the month preceding HZ infection. Recent use of cyclophosphamide or
methylprednisolone pulse therapy was recorded in 16(31%) and 5(9.6%) patients, respectively, four patients each
were receiving either azathioprine or mycophenolate mofetil.
CONCLUSION: HZ occurrence in this SLE cohort was 16.3%. Although 31% were on immunosuppressive
medications, HZ outcomes were generally favourable.
FUNDING: Lupus-Inspired Advocacy (LUISA) Project of Rheumatology Educational Trust Fund Inc. (RETFI)
P1-048 PROSTATIC ABSCESS AS A PRESENTING FEATURE OF PARADOXICAL TB-IRIS
C.H. Mak1, B. Achappa1, A. Shenoy1
1Medicine,
Kasturba Medical College Mangalore (Manipal University), MANGALORE DAKSHINA KANNADA DI, India
Paradoxical tuberculosis * immune reconstitution syndrome (pTB-IRIS) is a well recognized cause of clinical
deterioration in HIV tuberculosis (HIV-TB) co-infected individuals following initiation of antiretroviral therapy (ART).
pTB-IRIS is often reported as fever, lymphadenopathy & worsening of tuberculosis (TB) at a pre-existing site of
infection.
A 39 year old male, known case of retroviral disease was diagnosed with toxoplasmosis and TB meningitis. He was
started on antitubercular therapy (ATT) and steroids. Two weeks later, patient improved symptomatically and so ART
was initiated. He came back with complaints of fever with chills. Patient was adherent to treatment. His CD4 count
was 93 cells/mm3. Clinical examination revealed bilateral cervical lymphadenopathy. Investigations were sent to
evaluate the cause of fever but all were negative. Ultrasonography of abdomen & pelvis showed a heterogenous
hypoechoic lesion in the prostate with bulky right seminal vesicle, suggestive of an abscess. A transrectal ultrasoundguided aspiration of the prostate was carried out, which revealed acid fast bacilli. A complete drainage of the
tuberculous prostatic abscess was performed and patient was treated with steroids whilst continuing ART and ATT.
The patient improved symptomatically.
Although there was a strong clinical suspicion of pTB- IRIS, this case initially eluded the diagnosis and came across
as pyrexia of unknown origin because the site of worsening of TB if any, was not easily recognizable. Patient did not
report any complaints that could have suggest prostate involvement. This is probably the first reported case of pTBIRIS reported in literature which manifested as a prostatic abscess.
P1-049 Challenges in the management of concomitant TB arthritis and AVN in a patient with adverse drug
reaction to anti-Koch's medication-a case report
K. Tee1, A.D. Magbitang1, M.L. Tee1
1Section
of rheumatology, Philippine General Hospital, Manila, Philippines
BACKGROUND: Non-traumatic avascular necrosis(AVN) of the bone commonly presents as monoarthritis. While
corticosteroid still remains as one of the leading cause, several other factors have been associated with its
development. This includes systemic lupus erythematosus(SLE). A large proportion of patients with AVN benefits
from surgical procedures like core decompression. The presence of infection complicates its management.
A review of literatures showed only one case report of monoarticular tuberculous osteonecrosis diagnosed by
aspiration cytology.
Since tuberculosis and osteonecrosis are destructive but curable disease, early diagnosis and treatment is essential.
OBJECTIVE: To present a case of osteonecrosis of the hip and knee complicated by tuberculous arthritis of the knee
in a patient with SLE.
CASE: A 27 year old female who was diagnosed with lupus nephritis underwent 3 days methylprednisolone pulse
therapy. Lupus nephritis improved and SLE activity is clinically inactive for 2 years.
She developed insidious onset of monoarthritis of the left knee lasting for 6 months, with subsequent right hip pain of
1 week duration. She underwent arthrocenthesis of the left knee. Synovial fluid PCR was consistent with tuberculosis.
MRI of the left knee showed osteonecrosis and arthtritis. Radiograph of the right hip showed osteonecrosis. The
patient was given anti-Koch's regimen and iloprost infusion with significant clinical improvement.
CONCLUSION: This is the first reported case of a lupus patient with concomitant polyarticular osteonecrosis
complicated by monoarticular tuberculous arthritis. Medical treatment is effective for symptomatic management, but a
multidisciplinary approach is suggested for optimal outcome.
P1-050 HEPATITIS B-ASSOCIATED POLYARTERITIS NODOSA PRESENTING AS MONONEURITIS
MULTIPLEX: A CASE SERIES
M. Lazo1, J.P. Consignado1, S.T.V. Navarra1
1Section
of Rheumatology, St. Luke's Medical Center, Quezon City, Philippines
Background: Polyarteritis nodosa (PAN) is anANCA-negative systemic necrotizing vasculitis that typically affects
medium-sized muscular arteries, withoccasional involvement of small muscular arteries. Infectious etiologies suchas
Hepatitis B and C viruses and HIV are important in the pathogenesis of some ofthese cases. Hepatitis B-associated
PAN (HBV-PAN) has the same clinicalfeatures as non-HBV-associated PAN and typically occurs within four months
after
the
onset
of
HBV
infection.
Cases: We presenttwo cases of HBV-PAN seen over a 6-year period in our institution. The firstcase was a 38 year-old
male who presented with bilateral foot drop andhyperesthesia, weight loss, anorexia and fever, with EMG-NCV finding
of severedistal, asymmetrical, neuronal axonal loss type of sensorimotor polyneuropathy.The second case is a 43 yearold male with severe paresthesia and progressiveweakness of the distal upper and lower extremities. EMG-NCV
showed severe,diffuse, acute distal asymmetric sensorimotor axonal polyneuropathyaffecting the lower more thanthe
upper extremities. Both patients had chronic active Hepatitis B infectionby serologic testing. Sural nerve biopsy was
done in the 2 patients, results ofwhich were consistent with Polyarteritis Nodosa. Short-term steroid therapy,anti-viral
agents and serial plasma exchange provided improved motor strengthand eventually led to good outcome in these
patients.
Conclusion: HBV-PAN is an important consideration in patients presenting with vasculitic neuropathy, especially in
countries where prevalence of Hepatitis B infection is high. Corticosteroids, in adjunct with plasma exchange and antiviral therapy, are currently the cornerstone of treatment of HBV-PAN.
P1-051 HIGHER RESPONSE TO FEC PEPTIDE POOL STIMULATION IN RA PATIENTS UNDERGOING
GOLIMUMAB THERAPY
A. Khanniche1, P.H.D. Jiang1
1School
of medecine, Shanghai JiaoTong University, Shanghai, China
In order to investigate the susceptibility of RA patients to infections, we studied the functionality of virus specific
memory T cell upon stimulation with FLU-EBV-CMV peptide pools. RA patients included in this part are under TNF-a
inhibitor therapy (Golimumab) as we expect they may have more impaired memory T cells than patients without antiTNF- a therapy.
Methods: Upon isolation of PBMC from peripheral blood, we proceeded HLA-A2 phenotyping, intracellular staining
and flow cytometry.
Results:
Our results shown that patients under Golimumab demonstrated high sensitivity to stimulation with the FLU-EBVCMV peptide pool. Higher frequencies and diverse responses were observed in this group. For the first time, we
reported higher frequencies of CD8+ T cells specific for HLA-A2 restricted GILGFVFTL peptide in RA patients as
compared to the healthy control group.
Conclusion: This enhanced profile may be indicator for protection against infections and need further investigation.
Fig 1: Representative flow cytometry showing the response of CD8+ T cells to secrete IFN-g following
stimulation with FLU-EBV-CMV peptide pool in a RA patient.
Table1: Quality of the CD8+ T cells response to FEC stimulation
RA
HC
N
%
N
%
FLU-A2
10
43.47
5
38.46
FLU-B8
1
4.34
0
0
FLU-B27
1
4.34
0
0
EBV-B8
2
69
0
0
NR
9
39.13
8
61.53
All samples
23
100
13
100
Fig 2: CD8+ response to FLU-A2+ peptide stimulation in RA patients as compared to HC: (a) only HLA-A2+
samples were included, (b) only responders to FLU-A2+ stimulation are include
Poster Session: Session 1: Infection: the impact of parasites in the prevention of autoimmunity
P1-052 A CASE OF TETRAPLEGIA WITH SENSORY SPARING: SERIAL STUDIES
D. Kim1, B. Kim1, B. Park1
1Physical
Medicine & Rehabilitation, Korea university Ansan Hospital, Ansan-si, Korea
Introduction
Guillain-Barre syndrome (GBS) is an autoimmune disorder known as acute inflammatory demyelinating
polyradiculoneuropathy. However, the variants known as acute motor axonal neuropathy (AMAN) and acute motorsensory axonal neuropathy (AMSAN) also are well recognized.
Objective
.
Aims of study
.
Case report
A 28-year-old male presented with symmetric weakness of bilateral upper and lower extremities. He had an episode
of fever and diarrhea approximately 1 week before symptom development. Muscle strength on the Medical Research
Council (MRC) scale was grade 1 for the upper and lower limb muscles bilaterally. The sensory examination was
normal. Deep tendon reflexes were absent in the bilateral upper and lower limbs. In laboratory investigations,
erythrocyte sedimentation rate mildly increased and serum antibody to Campylobacter jejuni was positive. On day 2,
motor nerve conduction study (NCS) demonstrated no compound muscle action potentials (CMAPs) in the bilateral
median, ulnar, peroneal, and tibial nerves, while sensory nerve conduction studies were completely normal. He
underwent intravenous immunoglobulin treatment for 5 days was given. One week after onset, partial recovery was
observed in muscle strength. From twenty days after symptom onset, serial follow-up motor NCS revealed
progressively increased distal CMAP amplitude and definite partial conduction block in the forearm segment of most
of nerves. Needle EMG showed significant improvements in motor unit recruitment. Two months after onset, he was
discharged with mild weakness (MRC grade 4).
Methods
.
Result
.
Conclusion
Based on the clinical, laboratory, electrophysiological findings, subgroups of GBS should be defined. Viral infection
may precede acute motor axonal neuropathy causing rapid progressive ascending weakness with no sensory nerve
involvements.
Poster Session: Session 1: Liver and Autoimmunity
P1-053 A PATIENT WITH AUTOIMMUNE HEPATITIS AND AUTOIMMUNE PANCREATITIS; A CASE REPORT
OF AN UNCOMMON AUTOIMMUNE DISEASE ASSOCIATION
D. Dajcman1, N. Potocnik Dajcman2
1Gastroenterology
and gastrointestinal endoscopy, University Clinical Center, Maribor, Slovenia
Ambulatory unit, Children and adolescent psychiatry, Maribor, Slovenia
We report a case of an twenty years old Slovenian caucasian male with chronic asymptomatic pancreatic
hyperenzymemia, serum increased IgG4 level and with clinicaly and laboratory changes characteristic for
autoimmune hepatitis (AIH). Jaudince, characteristic elevation of liver enzymes, anti smooth muscle antibodies and
antinuclear antibodies were present. Since pancreatic hyperenzymemia is not frequently seen in patients with clinical
manifestation of AH, we investigated hepatic and pancreatic histopathlogy. Abdominal ultrasound, abdominal
computed tomography and magnetic resonance cholangiopancreatography showed no abnormalities in the
extrahepatic bile ducts and pancreatic duct, but there were diffuse tissue changes present in both organs on all
scans. An ultrasound-guided liver biopsy showed changes associated with autoimmune hepatitis: interface hepatitis,
lobular hepatitis and marked plasma cell infiltration. Endoscopic ultrasound (EUS)-guided fine needle aspiration
(FNA) of pancreatic tissue was performed for the evaluation of autoimmune pancreatitis (AIP); AIP was confirmed
based on the presence of lymphoplasmacytic sclerosing pancreatitis and immunoglobulin IgG4 positive plasma cells
in the infiltrate. For the definitive diagnosis, we used the Fukuoka 2010 International Consensus of Diagnostic Criteria
(ICDC) for autoimmune pancreatitis and existing international criteria for diagnosis of AIH. The patient was put on
steroid with satisfactory response on liver enzymes, jaundice and pancreatic hyperenzymemia. For maintenance
therapy and preventing relaps of both diseases we succsessfuly convert steroids to azathioprin therapy in maximal
dose daily. We concluded that there might exist an overlap syndrome between Type 1 AIH and AP which was in
Slovenia detected for the first time.
2Private
P1-054 FXR ACTIVATION ATTENUATES IMMUNE-MEDIATED LIVER INJURY VIA MYELOID-DERIVED
SUPPRESSOR CELLS
H. Zhang1, Z. Bian1, Q. Miao1, P. Invernizzi2, M. Gershwin3, X. Ma1
1Division
of Gastroenterology and Hepatology Ren Ji Hospital School of Medicine Shanghai Jiao Tong University,
Shanghai Institute of Digestive Disease, Shanghai, China
2Clinical Immunology, IRCCS Istituto Clinico Humanitas, Milan, Italy
3Division of Rheumatology Allergy and Clinical Immunology, University of California at Davis, Davis, USA
Background and Aims: FXR is a highly expressed hepatic nuclear receptor that exerts an important role in immune
regulation. We postulated that the cellular events that follow FXR activation include modulation of myeloid-derived
suppressor cells (MDSCs), a heterogeneous population with a remarkable ability to regulate innate immunity.
Methods: To address this issue, we induced hepatitis using both Con A and a-GalCer to monitor the natural history
of liver pathology in these two murine models of hepatitis with and without FXR activation, including use of animals
depleted of MDSCs and study of mechanisms of action using flow cytometry and adoptive cell transfer. We also
monitored the interactions of FXR activation and expression of PIR-B both in vivo and in vitro using luciferase
reporter and CHIP assays. Finally, we studied the potential of FXR activation to alter hepatic MDSCs homing.
Results: We report herein that FXR activation reduces the inflammatory injury induced either by a-GalCer or Con A;
such treatment expands CD11b+Ly6C+ MDSCs. The protective effect of FXR activation is dependent on expansion of
MDSCs, particularly liver CD11b+Ly6Chigh cells. Indeed, FXR activation enhances the suppressor function of MDSCs
through upregulation of PIR-B by binding the PIR-B promoter. FXR activation drives the accumulation of MDSCs to
liver through upregulation of S100A8. Conclusions: FXR activation facilitates homing and function of MDSCs, which
function as a critical negative feedback loop in immune-mediated liver injury. These novel mechanisms emphasize
the importance of defining liver lymphoid subpopulations.
Poster Session: Session 1: Mechanism of autoimmunity
P1-055 THE COMPLEX AND IRONIC RELATIONSHIP BETWEEN PD-1 AND TIM-3
Y. Lee1, K. Hyung1, Y. Moon1, M. Lee1, I. Lee1, B. Go1, K. Hwang1
1College
of Pharmacy, Chung-Ang University, Seoul, Korea
The immune responses of T cells are central in cell-mediated immunity and its activation is tightly regulated using costimulatory surface molecules CD28, CTLA-4, PD-1, Tim-3. Programmed cell death-1 (PD-1) and T cell
immunoglobulin mucin-3 (Tim-3) have been established as a negative regulatory molecule and plays a critical role in
immune tolerance.
PD-1+ Tim-3+ population is more dysfunctional than the PD-1+Tim-3-, PD-1-Tim-3- population and these phenotypes
are abundant in patients with cancer and chronic diseases.
Population classified by expression of PD-1 and Tim-3
There is solid evidence on the negative properties of these two molecules, however the mechanism behind the
synergistic amplification of the negative signal when these molecules expressed simultaneously, and the
interconnections between the signals of these two molecules have not been elucidated yet.
In this research, the expression of PD-1 and Tim-3 show inverse relationship in vitro culture system and in vivo tumor
model using wild-type, PD-1 Tg and PD-1 KO mice. In vitro culture, PD-1 expression of the PD-1 Tg, WT, PD-1 KO
was highly expressed in order, on the other hand, Tim-3 was expressed lower in that order. In vivo tumor model,
activated T cells persist in the tumor environment were more in PD-1 Tg mice, therefore tumor sizes after 30days
were smaller in PD-1 Tg compared to WT.
In conclusion, larger PD-1 but lower Tim-3 expression in PD-1 Tg suggests inverse relationship of these two
molecules and further study about these phenomenons are needed.
P1-056 MATERNAL METHYL-DONOR SUPPLEMENTATION INCREASES REGULATORY T-CELL MEDIATED
REDOX REMODELING
R. Yung1, S. Garg1
1Internal
Medicine, University of Michigan, Ann Arbor, USA
Very little is known about the role pre-natal environment plays in the pathogenesis of autoimmunity. Regulatory T
cells (Treg) maintain peripheral tolerance by inhibiting effector T (Teff) cells. Imbalance Treg function plays a unique
role in the development of autoimmune diseases. Previously, we exposed dams to a diet supplemented with methyl
donors (MS) or not (control) during mating and pregnancy, and found the MS diet suppresses the development of
atherosclerosis in F1 mice. Here, we studied the effects of these prenatal diets on Treg function in F1 mice. Dams
were fed either MS or control diets prior to mating, and during pregnancy and lactation. At 4 weeks of age, F1 mice
from both groups were either analyzed or placed on the standard NIH-31 chow and allowed to age. Prenatal MS diet
exposure is associated with lower Treg numbers in F1 mice, and this decrease is sustained into late life. Both FoxP3
mRNA and protein were decreased in mice on the MS diet, compared to the control diet. The difference in Treg
numbers is not due to differences in food consumption or weight gain between dams exposed to either prenatal diet.
Lower Treg cell numbers are associated with hyper-methylation of the FoxP3 promoter and enhancer regions.
Despite their lower numbers, Tregs from F1-MS mice produced more IL-10 and showed higher redox-dependent Teff
suppression, compared to Treg from F1-control mice. Together, these results show that a methyl-donor-rich prenatal
diet lowers total Treg numbers, though these cells maintain an anti-inflammatory phenotype.
P1-057 VARIETY OF ANTIBODIES TO THYREOPEROXIDASE IN THE AUTOIMMUNE DISEASES STUDY
A.V. Zubkov1, N.S. Kuzmina1
1Immunology,
Research Institute vaccines and serum of Mechnikov, Moscow, Russia
Background. The recent research of the thyroid autoimmune diseases concentrated on the detecting of the variety of
autoantibodies (Ab), directed against epitopes of the main thyreo autoantigenes: thyreoperoxidase (TPO) and
thyreotrop hormone receptor. The existing data strongly state that these epitopes are different for the patients with
different types of thyroid diseases.
Aim. The main aim of this work is the research Ab heterogeneity in case of GravesÕ and Hashimoto diseases using
monoclonal Ab (mAb) to TPO.
Materials and Methods. Ab heterogeneity has been researched by the competitive method of enzyme-linked
immunosorbent assay using mAb conjugates.
Results. The study of the human thyreoperoxidase Ab epitopes heterogeneity, extracted from the serum of the
patients with Graves' disease and Hashimoto disease has been conducted using mAb to TPO. It was shown, that 11
out of 23 used mAb to epitopes 1, 70, 82, 88, 2, 3, 10, 63, 45, 77 and 79 participate in the competition for the binding
centers of TPO and Ab, extracted from the Graves' diseased and Hashimoto diseased serum. The maxima of the
binding inhibition has been marked for the Ab, directed at the linear epitope 70 (TPO 710-720 ?.r.), Graves' disease *
50,1 ± 8,7 %, Hashimoto disease * 53,9 ± 9,2 %.
Conclusion. The linear epitopes of TPO, that cause the more 50 % of Ab to be produced by the patients with
different thyroid dysfunctions, have been detected. The future researches of the mAb heterogeneity will provide early
and effective diagnosis of these cases.
P1-058 HUMAN CD21LOWCD38LOW B CELLS ESCAPING NEGATIVE SELECTION REPRESENT A FOUNDER
OF AUTOREACTIVE B CELL POPULATION
S. Ruzickova1, M. Sedlackova2, H. Havelkova3, T. Kalina4, M. Vlkova5
1Laboratory
of Immunopathology and Immunotherapy, Institute of Biotechnology AS CR v. v. i., Prague 4, Czech
Republic
2Department of Rheumatology and Rehabilitation, Thomayer Hospital, Prague 4, Czech Republic
3Laboratory of Immunopathology and Immunotherapy, Institute of Biotechnology AS CR v. v. i., Brno, Czech Republic
4Department of Pediatric Hematology and Oncology, 2nd Faculty of Medicine Charles University, Prague 5, Czech
Republic
5Department of Clinical Immunology and Allergology, St Anne's Faculty Hospital, Brno, Czech Republic
In our previous study new CD19+CD21lowCD38lowIgM+CD27-CD24+/- B cell subpopulations have been observed in
patients with common variable immunodeficiency and controls (CO). The presence of B cells expressing VH4-34
gene known as coding for anti-dsDNA autoantibodies has been confirmed only in patients with CVID. The aim was to
search for such B cells in patients with rheumatoid arthritis (RA, N=2) and systemic lupus erythematosus (SLE, N=2),
and to analyse their molecular features using single-cell Ig specific RT-PCR.
All analysed B cells overexpressed IgM transcripts having mutational frequency in both CD24 + and CD24- RA/SLE B
cells decreased as compared to COs (1.74% and 1.47% vs. 2.35% and 1.96%). Most interestingly, the VH repertoire
was in RA/SLE B cells remarkably restricted to only 10 VH genes. In addition, the significant predominance of VH434 gene was found in patients« B cells of both subsets whereas in COs this gene was completely absent (0% vs.
43%; p=0.0092). The most surprising observation was extremely high degree of clonal relation of patients« B cells
while in COs this was not found (76% vs. 0%; p<0.0001). Moreover, over 60% of clonally related RA/SLE B cell used
VH 4-34 gene segment and their CDR3 regions were remarkably shortened (5-11 amino acids).
Observed data, namely prevalence of VH4-34 gene segment coding for autoantibodies might suggest the
susceptibility of these cells to escape negative selection and to serve as a source of autoreactive B cells under
autoimmune condition.
This work was supported by the grant NT/11414-5 from IGA MZ CR.
Poster Session: Session 1: Novel therapies and new biologics
P1-059 HUMORAL AND CELLULAR IMMUNE RESPONSES TO DIFFERENT FASCIOLA GIGANTICA ANTIGENS
I. Aly1, R. Zalat1, M. Younis1
1Immunoparasitology,
tbri, cairo, Egypt
In an attempt to develop a suitable vaccine against F. gigantica infection, two antigens were isolated and purified
from excretory/secretory (E/S) products, cystein protease and fatty acid binding proteins (CP and FABP) of the
parasite by immunoaffinity chromatography. Parasitological and immunological parameters were standardized. Noninfected (gp A) infected (gp B), immunized with CP (gp C), immunized with CP and infected (gp D), immunized with
FABP (group E) and immunized with FABP and infected (gp F). The mean worm burdens and bile egg count after
challenge were reduced significantly by 37.7% and 55.5%, respectively in rabbits vaccinated with CP. In contrast, low
significant reduction in worm burdens and bile egg count were observed in rabbits immunized with FABP after
challenge (23.5% and 35.7%, respectively). On the other hand, immunization of rabbits with CP or FABP induced a
significant expression of humoral antibodies and cytokines with higher level in case of CP than FABP. Among Ig
isotypes, IgG1 and IgG4 were most dominant in rabbits immunized with CP or FABP while, recording a low IgG2
level. On the other hand, among all cytokines IL-10 was most dominant in rabbits immunized with CP or FABP PI
suggesting also Th2 response, which could be another mechanism of immune response in rabbits infected with F.
gigantica. In conclusion, F. gigantica CP is a relevant candidate for vaccination against fascioliasis, while the level of
protection used by FABP may not appear sufficient enough to protect these ruminants against the deleterious effects
of Fasciola infection.
P1-060 EFFECT OF THAI HERBAL EXTRACTS CONTAINING ANTI-PSORIATIC ACTIVITY ON GENE
EXPRESSION OF EGR-1, IL-6 AND IL-8
C. Ronpirin1, T. Tencomnao2
1Preclinical
science, Thammasat University, Pathumthani, Thailand
for Excellence in Omics - Nano Medical Technology Development Project Department of Clinical Chemistry,
Chulalongkorn University, Bangkok, Thailand
The exact causes of psoriasis, a relatively common, chronic, inflammatory and hyperproliferative skin disease, are
not well understood, thus making it very difficult for therapy. The objective of this study was to investigate the
molecular effect of each of these following Thai herbal extracts, Alpinia galanga L. (rhizome), Curcuma longa L.
(rhizome) and Annona squamosa L. (leaf), containing anti-psoriatic activity on the mRNA expression of Egr-1, IL-6
and IL-8, the biomarkers believed to be upregulated in psoriatic skin, thus considering as one of essential
determinants of psoriasis. Using HaCaT keratinocyte cell line as a model, the mRNA expression was evaluated by
quantitative real time PCR. We found that none of the ethanolic extracts in this study could modulate the expression
of all analyzed genes. Nevertheless, the extract of Annona squamosa L. at 0.25IC50 could significantly suppress the
expression of IL-8 mRNA (P<0.05). The extract of Alpinia galanga L. at IC50 could significantly reduce the expression
of IL-6 mRNA (P<0.05). Therefore, the result suggests that the ethanolic extracts of Annona squamosa L. (leaf) and
Alpinia galanga L. (rhizome), contain certain constituents capable of downregulating these proinflammatory cytokines.
This information is useful for further studies, which may provide insight into understanding therapeutic options in the
future.
2Center
P1-061 ANTI-INFLAMMATORY EFFECT OF HOUTTUYNAI CORDATA THUNB. LEAF EXTRACTS ON HUMAN
KERATINOCYTE CELLS
C. Ronpirin1, T. Tencomnao2
1Preclinical
science Faculty of Medicine, Thammasat University, Pathumthani, Thailand
for Excellence in Omics - Nano Medical Technology Development Project Department of Clinical Chemistry,
Chulalongkorn University, Bangkok, Thailand
Psoriasis is a chronic, non-contagious autoimmune disease that affects the skin and joints. Currently, scientists have
attempted to find a therapeutic approach with highest efficacy. In particularly, attention has been focused on the
applications of medicinal herbs for treatment and prevention of psoriasis. In this study, we aimed at evaluating antiinflammatory and anti-psoriatic activities of Houttuynai cordata Thunb. leaves extracted with 2 different methods. The
fist technique was exhaustive extraction using petroleum ether, dichloromethane and ethanol. The second one was
maceration with ethanol. Based on MTT assay, we found that Houttuynai cordata Thunb. leaf dichloromethane
extract (100, 50, 25, 12.5 and 6.25 µg/mL) and petroleum ether extract (100, 50, 25 and 12.5 µg/mL) exhibited
cytotoxic effect on cultured HaCaT cells. According to our ELISA assay, Houttuynai cordata Thunb. leaf
dichloromethane extract (100 and 6.25 µg/mL) led to apoptosis of HaCaT. With regard to anti-inflammatory activity
using ELISA for pro-inflammatory cytokines, interleukin-6 and interleukin-8, Houttuynai cordata Thunb. leaf petroleum
ether extract (12.5 and 6.25 µg/mL) could suppress the production of interleukin-6, while other Houttuynai cordata
Thunb. leaf extracts did not affect the expression of pro-inflammatory cytokines in HaCaT cells. Different extraction
methods certainly resulted in extracts with distinct biological activities. It should be worth mentioning that other
techniques should be employed for further studies. Nevertheless, the result of this current investigation is important
with respective to new findings that may be useful for psoriatic patients in the future.
2Center
P1-062 GAS-THERAPY IN RHEUMATOID ARTHRITIS (RA) TREATMENT: WHEN WEST MEETS EAST.
M. Gajewski1, S. Maslinski2, P. Rzodkiewicz1, A. Paradowska-Gorycka1, E. Wojtecka-Lukasik1
1Department
of Biochemistry and Molecular Biology, Institute of Rheumatology, Warsaw, Poland
of General and Experimental Pathology, Medical University of Warsaw, Warsaw, Poland
It is possible to regulate the activity of connective tissue (CT) by acupuncture by acting on to ãacupointsÓ (sites of
greatest CT accumulation). High concentrations of acetylcholine - neutrotransmitter of cholinergic system were found
within the bondary of most acupuncture points. It is suggested that autonomic nervous system, vagus nerve can be
influenced by acupuncture. This treatment increases the efficiency of transport of gasotransmitters: such as carbon
monoxide, nitric oxide and hydrogen sulfide which are produced endogenously, play a roles neurotransmitters and
inhibits energy production, conversely to exogenously oxygen enhanced this reaction. The famous ãQi energyÓ
would be nothing else than a state of molecular equilibrium ( redox potential) that may shift towards either activation
(oxygen * Yang) or inhibition (gasotransmitters * Yin).
2Department
Redox state can regulate inflammatory cells differentiation. In synovial fluid neutrophils differentiation toward to
highly-specialized ãmicrophagesÓ was observed. These cells are responsible for major injury in the joint and can
disrupt the capacity of endothelial progenitor cells to differentiate into mature endothelial cells (bloking of
neovascularization of ãpannusÓ). Moreover, neutrophiles might be responsible for controlling migration and
activation of lymphocytes and monocytes. Targeting neutrophils seems to be very promising therapeutic metod.
Taken together, understanding of the complex interactions between CT and vagus nerve modulation
(ãacupunctureÓ), gases neurotransmitters and redox status of inflammatory cells (ãQi energyÓ) provide valuable
opportunities for the development of novel therapeutic strategies for RA. Once again western medicine reveals
knowledge that has been known in an eastern medicine ages before.
P1-063 INDUCTION OF APOPTOSIS BY HEP88 MONOCLONAL ANTIBODY IN HEPATOCELLULAR
CARCINOMA
K. Aree1, T. Mitupatum2, S. Puthong3, P. Rojpibulstit2
1Division
of Microbiology and Immunology Department of Preclinical Science, Faculty of Medicine Thammasat
University, Pathumthani, Thailand
2Division of Biochemistry Department of Preclinical Science, Faculty of Medicine Thammasat University,
Pathumthani, Thailand
3The Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok, Thailand
Hepatocellular carcinoma is a leading cause of cancer-related death worldwide. Hep88 mAb, an antibody against
tumor-associated antigen in hepatocellular carcinoma cell line established from a Thai patient, has been previously
shown to induce cancer cell death. These dying cells exhibited morphological characteristics of apoptosis-like
programmed cell death (PCD), including cytoplasmic vacuolization and dilatation of mitochondria and endoplasmic
reticulum. In the present study, effects of Hep88 mAb on cell cycle progression and apoptosis induction in human
hepatocellular carcinoma cell line, HepG2, were examined. Growth-inhibitory activity of Hep88 mAb on HepG2 cell
line was associated with G2/M cell cycle arrest. In addition, Hep88 mAb induced a significant increase in the sub-G1
peak, indicating an apoptotic cell population. The apoptosis-inducing effects of Hep88 mAb were further investigated
using Annexin V-FITC/propidium iodide staining and flow cytometry. Early apoptotic cells (Annexin V single positive
cells) were increased by Hep88 mAb treatment in a dose-and time-dependent manner. Although it has been
suggested that apoptosis-like PCD is involved in anti-tumor activity of Hep88 mAb, the results from this study
demonstrated that Hep88 mAb induced Hep G2 cell line to undergo cell cycle arrest and subsequent apoptosis.
Therefore, further investigation of the cellular and molecular mechanisms underlying anti-cancer activities of Hep88
mAb as well as the potential uses as an anti-neoplastic agent against hepatocellular carcinoma is required.
P1-064 IMMUNOSUPPRESSIVE ACTIVITY OF A NOVEL WATER-SOLUBLE BENZOTHIAZOLE DERIVATIVE,
BD926, ON T CELL PROLIFERATION IN VITRO AND IN VIVO
Y. Liu1, Q. Zou1
1Department
of Immunology, Chengdu Medical College, Chengdu, China
Aberrant T cell responses mediate a variety of autoimmune diseases, including rheumatoid arthritis, systemic lupus
erythematosus, multiple sclerosis and ulcerative colitis. Immunosuppressants are widely used for the treatment of T
cell-mediated autoimmune diseases. However, because of the toxicity and tolerance of these drugs, it is important to
keep looking for novel immunosuppressants. Here, a series of novel benzothiazole derivatives were synthesized and
screened for the immunosuppressive activity. A water-soluble benzothiazole derivative, BD926 had the most effective
activity among these compounds. Treatment with BD926 significantly inhibited mouse and human T cell proliferation
stimulated either by anti-CD3/anti-CD28 mAbs or by alloantigen in a dose-dependent manner measured by flow
cytometry in vitro. We did not observe obvious cytotoxicity of BD926 against human resting na•ve T cells, IL-4
treated-activated T cells and fibroblast-like synoviocyte determined by CCK-8 assay. Furthermore, BD926 did not
inhibit IL-2, IL-4 and IL-10 secretion, but inhibited IFN-g, IL-6 and IL-17 production measured by ELISA and induced
cell cycle arrest at G0/G1 phase in activated T cells determined by flow cytometry. Interestingly, AKT and p70S6K
phosphorylation was not inhibited by BD926 in IL-2-induced T cells assessed by western blot. Finally, administration
of BD926 mitigated T cell-mediated delayed-type hypersensitivity in mice in a dose-dependent manner in vivo.
Collectively, these data suggest that BD926 inhibits T cell proliferation and may be used as a lead compound for the
design and development of new immunosuppressant for the intervention of autoimmune diseases.
P1-065 LONG-TERM EFFECTIVENESS OF AUTOLOGUS WHOLE BLOOD INJECTIONS TO PATIENTS WITH
CHRONIC SPONTANEOUS URTICARIA WITH POSITIVE AUTOLOGOUS SERUM SKIN TEST
L. Na1
1Dermatology,
Southwest Hospital Third Military Medical University, Chongqing, China
Background and Aims: To evaluate the clinical effects of autologus whole blood injections (AWBI) on patients with
chronic spontaneous urticaria (CSU) with positive autologous serum skin test (ASST).
Methods: On the basis of evaluation of clinical history, 160 patients with positive ASST but negative skin prick test
(SPT) to common allergens were randomly subgrouped into AWBI-treatment group and placebo-control group by
intramuscular injection once a week for twelve times. At the meantime, gradually tapered oral loratadine. Both of the
two groups were administered and the dosages were off after disappearance of symptoms of urticaria. The urticaria
activity score (UAS), Dermatology Life Quality Index (DLQI) and the total required antihistaminics at the end of the
third, sixth, ninth and twelfth month after initial treatment were evaluated.
Results: Compared with that in control group, the scores of UAS and DLQI in AWBI-treatment group were gradually
decreased at the third(70% vs. 40%, 83% vs. 50%), sixth (88% vs. 57%, 93% vs. 60%), ninth(90% vs. 60%, 95% vs.
62%)and twelfth month (94% vs. 68%, 96% vs. 65%) respectively after initial treatment. Furthermore, the total doses
of required antihistaminics were also significantly reduced compared with that in control group at different time points
(9.06 vs 15.28 pills, 3.71 vs 13.21 pills, 1.83 vs 11.31 pills, 1.35 vs 10.98 pills).
Conclusions: Combination with oral antihistamine and AWB injections could not only improve urticarial symptoms
and the disease activity, but also could remarkably reduce the total doses of antihistaminics which are essential to
control clinical symptoms.
P1-066 NOVEL SMALL MOLECULE INHIBITORS OF FCGRIIA FOR AUTOIMMUNE DISEASE
D. Cox1, M. Brennan1, M. King1, M. Adamo1
1Molecular
& Cellular Therapeutics, Royal College of Surgeons in Ireland, Dublin, Ireland
Background. FcgR is a family of receptors that specifically bind IgG and its most important member FcgRIIa plays a
key role in phagocytosis. In autoimmune diseases there is excessive formation of immune complexes that then binds
FcgRIIa and plays an important role in pathogenesis. In animal models of rheumatoid arthritis activation of monocyte
FcgRIIa leads to over production of TNFa, which drives the inflammation of the joints. There is also evidence of a role
for FcgRIIa in autoimmune thrombocytopenia (ITP) where antibodies to FcgRIIa can prevent the thrombocytopenia.
Currently there are no FcgRIIa antagonists available for clinical use although pooled IgG is known to mediate its
effects in part by inhibition of FcgRIIa.
Aims. We describe the development of novel small molecule inhibitors of FcgRIIa.
Methods. MOE was used for virtual high-throughput screening (VS) of ZINC database. Platelet adhesion to IgG was
used as the high-throughput screen (HTS) since FcgRIIa is their only Fc receptor.
Results. Using the crystal structure of FcgRIIa we developed a pharmacophore for IgG binding. The hits identified
from the VS screen were tested for activity in the HTS and a chemical template identified. Over 100 derivatives were
synthesized and screened in the HTS. The most potent derivatives have IC 50 values of 1 mM in the HTS. Similar
activity was also found in functional assays such as heat-agglutinated IgG-induced platelet aggregation.
Conclusions. These compounds have the potential to be lead compounds for a new generation of drugs for
autoimmune disease such as rheumatoid arthritis and ITP.
P1-067 THE CYTOKINE MIDKINE AND ITS RECEPTOR RPTPZ REGULATE B CELL SURVIVAL IN A PATHWAY
INDUCED BY CD74
S. Cohen1, O.Y. Shoshana1, E. Zelman-Toister1, N. Maharshak1, I. Binsky-Ehrenreich1, I. Hazan-Halevy2, Y.
Herishanu2, L. Shvidel3, M. Haran3, L. Leng4, R. Bucala4, S. Harroch5, I. Shachar1
1Immunolgy,
Weizmann Institute of Science, Rehovot, Israel
Tel Aviv Sourasky Medical Center, Tel Aviv, Israel
3Hematology, Kaplan Medical Center, Rehovot, Israel
4Medicine, Yale University School of Medicine, New Haven, USA
5Neuroscience, Institut Pasteur, Paris, France
Lasting B cell persistence depends on survival signals that are transduced by cell surface receptors. In this study, we
describe a novel biological mechanism essential for survival and homeostasis of normal peripheral mature B cells
and chronic lymphocytic leukemia cells, regulated by the heparin-binding cytokine, midkine (MK), and its
proteoglycan receptor, the receptor-type tyrosine phosphatase ζ (RPTPζ). We demonstrate that MK initiates a
signaling cascade leading to B cell survival by binding to RPTPζ. In mice lacking PTPRZ, the proportion and number
of the mature B cell population are reduced. Our results emphasize a unique and critical function for MK signaling in
the previously described MIF/CD74-induced survival pathway. Stimulation of CD74 with MIF leads to c-Met activation,
resulting in elevation of MK expression in both normal mouse splenic B and chronic lymphocytic leukemia cells. Our
results indicate that MK and RPTPζ are important regulators of the B cell repertoire. These findings could pave the
way toward understanding the mechanisms shaping B cell survival and suggest novel therapeutic strategies based
on the blockade of the MK/RPTPζ-dependent survival pathway.
2Hematology,
Poster Session: Session 2: Old &amp; new autoimmune diseases
P2-001 MYCOPHENOLIC ACID FOR HYPER-IGE SYNDROME
A. Widhani1, N. Sukmana1
1Internal
Medicine Department, Faculty Medicine Universitas Indonesia, Jakarta, Indonesia
Background. Hyper-IgE syndrome (HIES) is a multisystem disorders related to eczema/atopic like dermatitis,
recurrent pulmonary and skin infections, eosinophilia, and increased in serum IgE level. Patients with HIES have
increased risk of autoimmune diseases and malignancies. The management of HIES is difficult. Rituximab and
omalizumab had been investigated but no significant clinical improvement observed.
Aims. Here we reported eight cases of HIES treated with mycophenolic acid to study its efficacy.
Methods. We examined a case series of eight patients. Clinical manifestations, clinical improvement, and IgE level
were studied.
Results. From 2011-2012 there were eight cases (4 males and 4 females) of HIES treated with mycophenolic acid.
All cases had dermatological manifestation (6 cases with erythroderma and 2 cases with atopic like dermatitis) with
half cases had history of drug allergy. Pulmonary infiltrate was found in 3 cases. There were renal involvement in 2
cases (nephrotic syndrome) and gastrointestinal manifestations in 2 cases (chronic illeitis and multiple esophageal
ulcer). Patients treated with steroid and mycophenolic acid had IgE more than 2000 ng/mL, normal CD4 cell count,
and no sign of malignancy or severe infection. Seven cases showed clinical improvement after one month treatment
and one case needed increasing dose. In all cases, after three months treatment, the IgE level decreased.
Conclusion. Mycophenolic acid proved to be safe and effective in treating some autoimmune diseases, can
decrease IgE level and improve clinical manifestations in HIES. A clinical trial with more subjects is needed to prove
its efficacy and safety in HIES.
P2-002 HYPER-IGE SYNDROME PRESENTING AS MULTIPLE DRUG ALLERGY, CHRONIC DIARRHEA, AND
HYDROPNEUMOTHORAX- A CASE STUDY
A. Widhani1, N. Sukmana1, C.M. Rumende2, W.K. Budianti3, I. Wuryantoro4
1Allergy
and Clinical Immunology Division Internal Medicine Department, Faculty of Medicine University of Indonesia,
Jakarta, Indonesia
2Pulmonology Division Internal Medicine Department, Faculty of Medicine University of Indonesia, Jakarta, Indonesia
3Dermato-Venerology Department, Faculty of Medicine University of Indonesia, Jakarta, Indonesia
4Thoracic Cardiovascular Surgery, Faculty of Medicine University of Indonesia, Jakarta, Indonesia
Background. Hyper-IgE syndrome (HIES) is multisystem disorder which is related to increase in the serum IgE,
dermatitis, eosinophilia, and recurrent pulmonary or skin infections. Patients with HIES have increased risk of
autoimmune diseases.
Aims. We aimed to describe a case of HIES with rare manifestations: multiple drug allergy, chronic diarrhea, and
hydropneumothorax and evaluate outcome of treatment with mycophenolic acid.
Methods. We report a case of 42 years old man with chronic diarrhea and pneumonia been treated with some
antibiotics for 4 months without any improvement. Diagnostic procedures were done and revealed HIES.
Results. The biopsy from intestinal mucosa showed active chronic illeitis, with negative congo red staining. The
antinuclear antibody titer was more than 1/1000 with coarse speckled pattern. Anti-dsDNA, anti-Sm, anti-SSA, antiSSB, and anti-Rib-P protein were negative. CD4 cell count was normal. During hospitalization he developed
spontaneous left hydropneumothorax. The pleural fluid was exudates and adenosine deaminase was increased. The
patient was given antituberculosis drugs, but no improvement. The lung remained unexpanded. Later he developed
maculopapular rash. The anti-TB drugs were then stopped. The laboratory results revealed increased serum IgE and
eosinophilia. The patient was diagnosed with HIES. He was given intravenous metyhlprednisolone and mycophenolic
acid. After the immunosuppressive drugs were started, the rash, diarrhea, and left hydropneumothorax were
resolved. The serum IgE level decreased two months after therapy
Conclusions. This case illustrates rare manifestations of HIES. It also shows that mycophenolic acid can improve
symptoms and decrease IgE level in HIES.
P2-003 A CASE OF IGG4-RELATED XANTHOMATOUS HYPOPHYSITIS
E. Csajbok1, S. Magony1, K. Sepp1, Z. Valkusz1, P. Barzó2, L. Tiszlavicz3
11st
Department of Internal Medicine Endocrine Unit, University of Szeged, Szeged, Hungary
of Neurosurgery, University of Szeged, Szeged, Hungary
3Department of Pathology, University of Szeged, Szeged, Hungary
Background: Hypophysitis is an inflammatory disease of the pituitary that may mimic tumors. Primary hypophysitis
has been classified as lymphocytic (LH), granulomatous (GH), and xanthomatous (XH). It has been recently
proposed to be an IgG4-related autoimmune disease (serum IgG4 concentration >135mg/dl).
2Department
Case description: We report on a case of a XH initially diagnosed as pituitary adenoma. A 23-year-old men
suffered from typical cluster type headache. Two years after the first symptoms, diabetes insipidus occured. All the
anterior pituitary hormone levels were normal thought the testosterone level was low. Sella MRI scan depicted a 17
mm inhomogenous mass. After the transphenoidal surgery the pituitary tissue was showed by accumulation of foamy
cells and xanthomatous epithelioid cells. After stopping the preoperative hydrocortisone therapy severe cluster type
headache returned. The endrocrine work up revealed hypadrenia (morning cortisol: 96 nm/l, ACTH:3.38 pm/l),
hypothyroidism (ft4:10.5 pm/l), hypogonadism (testosterone: 3.44 nm/l) with FSH:3,3 mIU/l and LH:2,8 mIU/l.
Hydrocortisone, levothyroxine and testosterone were stepwise reintroduced. During the follow-up we could stop
hydrocortisone, levothyroxine, whereas he has permanently required desmopressin and testosterone substitution.
The cluster type headache attacks dissappeared with glucocorticoid administration. The serum IgG4 concentration
was 815 mg/l suggesting the hypophysitis to be IgG4- related.
Conclusion: We describe an unusual case of IgG4 related xanthomatous hypophysitis causing cluster type
headache permanenty requiring ddAVP (desmopressine) and testosterone supplementation however, without need
for maintenance medication with hydrocortisone and levothyroxin. In periods of headaches the patient requires
glucocorticoids supporting the possible autoimmune origin of the disease.
Poster Session: Session 2: Pregnancy in autoimmune disease
P2-004 PREGNANCY-INDUCED CHANGES IN REGULATORY NETWORK OF NKT AND T REGULATORY
CELLS (TREGS) IN AUTOIMMUNE THYREOID DISEASE
I. Mrakovcic-Sutic1, T. Bogovic-Crncic2, S. Grbac-Ivankovic2, A. Bulog3, V. Micovic3, Z. Baricev-Novakovic4, I. Sutic5,
V. Pavisic5
1Department
2Department
of Physiology and Immunology, Medical Faculty Univercity of Rijeka, Rijeka, Croatia
of Nuclear Medicine Clinical Hospital Centre Rijeka Croatia, Medical Faculty Univercity of Rijeka, Rijeka,
Croatia
3Department of Public Health Medical Faculty University of Rijeka Croatia, Medical Faculty Univercity of Rijeka,
Rijeka, Croatia
4Department of Family Medicine Medical Faculty University of Rijeka Croatia, Medical Faculty Univercity of Rijeka,
Rijeka, Croatia
5Medical Faculty University of Rijeka Croatia, Medical Faculty Univercity of Rijeka, Rijeka, Croatia
Background and aims: autoimmune thyroid disease (AITD) represent a group of disorders of organ function that
occur as a result of immune responses directed against its own thyroid follicular cells in the presence of circulating
antibodies specific for thyroid antigens, following with activation of cellular immune response, releasing of cytokines,
lymphocyte infiltration and cell destruction. AITD includes two main clinical entities: Graves' disease and Hashimoto
thyroiditis. They affect often women of reproductive age. During healthy pregnancy predominant is Th2 over Th1
immune response, which explains the improvement of autoimmune disease during pregnancy, while after delivery
because of the changes in Th1/Th2 ratios often leads to deterioration of AITD. Regulatory network of NKT and Tregs
seems to play an important role in mediating maternal tolerance to fetus.
Methods: We investigated the role of NKT and Tregs in presence of ATD in pregnancy and in postpartum period in
women with hormonal status determination and with positive titer of thyroid antibodies and autoantibodies and
compared them with healthy pregnant and not pregnant women, using flow cytometry analysis.
Results: cells of innate immunity: NKT and Tregs are increased in healthy pregnancies, in pregnancies with hypo-and
hyperthyroidism, and postpartum in examine groups. NKT cells are decreased in pregnant women with positive
antibodies indicating a lost of NKT cellsÕ protective effect in pregnancy with positive antibodies.
Conclusion: Pregnancy and early postpartum period have a great effect on interaction of innate immunity and the
function of the thyroid gland. During AITD changes are more pronounced, especially postpartum.
P2-005 DIFFICULT LUPUS NEPHRITIS IN PREGNANCY
M. Lazo1, J.J. Lichauco1
1Section
of Rheumatology, St. Luke's Medical Center, Quezon City, Philippines
Introduction: Systemic lupus erythematosus (SLE) is an autoimmune disease that occurs frequently in women of
childbearing age. Although fertility is not compromised, pregnancies among SLE patients are commonly associated
with complications. We describe a case of a 30 year old Filipino female who had an unplanned pregnancy during
active lupus nephritis (Type IV). She was treated with monthly Methylprednisolone pulse therapy but eventually
delivered via caesarian section (C- section) due to intrauterine growth retardation (IUGR) at 35 weeks AOG.
Case report: A 30 year-old female (G3P3) was diagnosed with Type IV Lupus Nephritis in 2005. She received
induction treatment with IV Cyclophosphamide for 6 months followed by Mycophenolate mofetil at 2 gms/day for
maintenance treatment. She continued to have active disease and subsequently received 2 doses of Rituximab 1 gm
IV with 500 mg/m2 IV Cyclophosphamide. She achieved clinical remission for 3 months but had an unplanned
pregnancy during recurrent active kidney disease. Proteinuria with active urinary sediments were observed despite IV
pulse methylprednisolone therapy. At 35 weeks AOG, abdominal ultrasound showed asymmetric IUGR requiring Csection.
Discussion and Conclusion: Pregnancy in women with lupus nephritis has an increased risk of fetal loss and lupus
flares. Risk factors associated with poor pregnancy outcomes include a serum creatinine of >2.8mg/dl, active
nephritis, hypertension during the first trimester, and presence of anti-phospholipid antibodies. Since treatment
options for active lupus during pregnancy are limited, this case emphasizes the importance of planned pregnancies
during lupus disease quiescence to provide the best maternal and fetal outcome.
P2-006 TREATMENT OF INSULIN ALLERGY DURING PREGNANCY WITH INSULIN ANALOG
S. Hawisa1, K. Hawisa2
1Microbiology
and Immunology, School of Medical Sciences, Tripoli, Libya
and gynaecology, Zawia teaching hospital School of Medical Sciences, Zawia, Libya
Allergy to insulin is rare due to the introduction of human insulin and its analogues, and currently reported in less than
1-2% of diabetic patients. Although, immunologic responses to insulin in the form of IgG and IgE antibodies are
common clinical problems related to these immunologic reactions are uncommon. Usually, the allergic reactions are
limited to the site of insulin injection and systemic, potentially life-threatening reactions such as anaphylaxis have
been reported infrequently. However, treatment of diabetics with insulin allergy during pregnancy could be a real
challenge for the treating physician in countries with the lake of experience with the use of new insulin alogs.
Treatment options for insulin allergy include symptomatic therapy with antihistamines, use of an alternative insulin
preparation, use of oral hypoglycaemic agents, insulin desensitization using small doses of insulin subcutaneously,
and pancreatic transplantation for severe resistant cases. However, management in pregnant women is more difficult
as treatment options are limited. A 29-year-old pregnant woman with type 1 diabetes developed generalized
erythematous skin rash with marked pruritus following insulin administration. She has a strong family history of
diabetes; her mother, all her sisters and her brother are diabetics, her mother has allergy to insulin (protamine). Due
to insulin allergy, the patientÕs diabetes was controlled by switching to insulin Lispro with no adverse effects. Her
antenatal follow up was regular but needed frequent visits; she had a successful pregnancy outcome with no
congenital abnormalities in the baby.
2Obstetrics
Poster Session: Session 2: Prognostic markers
P2-007 VASOACTIVE INTESTINAL PEPTIDE RECEPTORS IN CHRONIC FATIGUE SYNDROME.
K. Fuller1, E.W. Brenu1, T.K. Huth1, S.L. Hardcastle1, S. Johnston1, D.R. Staines2, S. Marshall-Gradisnik1
1National
Center for Neuroimmunology and Emerging Diseases, Griffith University, Southport, Australia
Coast Public Health Unit, Queensland Health, Robina, Australia
Background and Aims: Vasoactive neuropeptides such as vasoactive intestinal peptide (VIP) and pituitary
adenylate cyclase activating polypeptide (PACAP) are known to play a critical role in immune regulation. These
activate VIP and PACAP receptors including VPAC1, VPAC2 and PAC1, which stimulate adenylyl cyclase activity.
Chronic Fatigue Syndrome (CFS) is a complex disorder which is known to possess a significant immune impairment
component. To date, only one study has shown an increase in VPAC2 on CD4 +T cells in CFS patients. Hence, the
study aims to assess the expression of VPAC1, VPAC2 and PAC1 on B-cells in CFS patients.
2Gold
Methods: Peripheral blood samples were collected from patients fulfilling the 1994 Centers for Disease Control and
Prevention, and the 2011 International Consensus Criteria case definitions for CFS, and from non-fatigued control
participants. Isolated peripheral blood mononuclear cells, were stimulated with 1µg of lipopolysaccharide, or PMA,
and cultured for 48 hours. Samples were then stained with the appropriate monoclonal antibodies and analysed on
the flow cytometer to determine the levels of VIP and PACAP receptors on CD19 + B-cells.
Results: CFS patients displayed differential levels in the expression of VIP and PACAP receptors on the surfaces of
CD19+ B-cells following stimulation.
Conclusion: It has been previously shown that VPAC2 receptors are elevated on the surfaces of CD4 + T-cells in
CFS patients. Hence, this study has demonstrated a role of VIP and PACAP receptors in modulating B cells and
other immune processes.
Poster Session: Session 2: Rheumatoid Arthritis
P2-008 ASSOCIATION OF MIR-181A AND IL-2 WITH THE DEVELOPMENT OF ANKYLOSING SPONDYLITIS
C. Chou1, J. Zhan2, T. Tseng2, C. Huang1, J. Wei1, J. Wang1, R. Wong2
1Institute
of medicine, Chung Shan Medical University, Taichung, Taiwan
of Public Health, Chung Shan Medical University, Taichung, Taiwan
Background and aims:The dysfunction of central tolerance can cause the development of autoimmune disease, and
microRNA (miR)-181a plays a critical role in central tolerance. Higher miR-181a expression could induce the T cell
receptor (TCR) signaling, which might induce higher expression of activated T cell. In addition, higher miR-181a
expression also could affect the interlukin (IL)-2 expression which influenced negative selection of T cell. Therefore,
we designed a hospital-based case-control study to evaluate the effects of miR-181a and IL-2 in the development of
ankylosing spondylitis (AS).
Methods:Whole blood miR-181a expression was detected by real-time quantitative polymerase chain reaction among
128 AS patients and 128 age and gender-matched controls. Serum IL-2 expression was measured using enzymelinked immunosorbent assay.
Results:Our results observed that AS patients had the significantly increased IL-2 expression than healthy controls (P
< 0.01). A significantly positive correlation of miR-181a and IL-2 was observed in AS patients (r = 0.32, P < 0.01).
Further, subjects with both of higher miR181a expression and higher IL-2 expression had 2.87-fold increased risk for
AS development (95% confidence interval = 1.46-5.62) compared to those with both of lower miR-181a expression
and lower IL-2 expression. In addition, AS patients with higher miR-181a expression had the increased ESR level
than those with lower miR-181a expression (P = 0.03). AS patients with higher IL-2 expression also had the
significantly higher risk for peripheral arthritis than those with lower IL-2 expression (OR = 3.95, 95% CI = 1.0114.74).
Conclusions:In conclusion, expressions of miR-181a and IL-2 might be correlated with AS development and clinical
characteristics.
2Department
P2-009 SYNOVIAL APOPTOSIS IS INVOLVED IN THE REGULATION OF AUTOIMMUNE ARTHRITIS BY GRAPE
SEED PROANTHOCYANIDIN
J. Kim1, H. Jeong2, H. Lee3, G. Choi1, S. Kim1
1Division
of Rheumatology Department of Internal Medicine, Keimyung University Dongsan Medical Center, Daegu,
Korea
2Department of Internal Medicine, Keimyung University Dongsan Medical Center, Daegu, Korea
3Microbiology, Keimyung University School of Medicine, Daegu, Korea
Background and aims: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by the synovial
inflammation, proliferation and bone destruction. Grape seed proanthocyanidin extract (GSPE), which is the
antioxidant derived from grape seeds, has been reported to have a variety of potent biologic functions. This study
was aimed to determine the effect of GSPE on arthritic inflammation and to reveal the mechanism by which GSPE
regulates immune response.
Methods: Mice with collagen-induced arthritis (CIA) were treated with 100mg/kg GSPE or saline via intraperitoneal
injections five times weekly for 2 weeks. Clinical assessment of arthritis and histologic analysis were performed.
Western blot was used for evaluating the expression of proapoptotic proteins.
Results: GSPE treatment ameliorated the arthritis symptoms of CIA. GSPE-treated mice showed a reduction in the
serum levels of pro-inflammatory cytokines including IL-6, TNF-α, and IL-17. The joints of the GSPE-treated mice
exhibited decreased levels of inflammatory cell infiltration, synovial proliferation, and articular cartilage destruction.
Radiographic evaluation of hind paws also revealed the reduction of the degree of bone resorption in the GSPEtreated mice. The expression of the proapoptotic proteins such as caspase 3 and p53 was increased in the synovium
of the GSPE-treated mice.
Conclusions: Our findings demonstrate that GSPE has an anti-inflammatory effect on murine autoimmune arthritis
by promoting apoptosis of synovial cells. These results suggest that GSPE might be a candidate therapeutic agent for
RA.
P2-010 INCREASED LYMPHOCYTE INFILTRATION IN RHEUMATOID ARTHRITIS IS CORRELATED WITH AN
INCREASE IN LINEAGE-CD4-CD127+ CELLS
S. Kim1, Y.E. Lee1, W.J. Jung1, G.G. Song2, M.Y. Kim1
1Bioinformatics
and Life Science, Soongsil University, Seoul, Korea
Korea University Guro Hospital, Seoul, Korea
The current study explored the relationship between lineage-CD4-CD127+ cells and lymphocyte infiltration in
rheumatoid arthritis. A lineage-CD4-CD127+ cell is positive for CD117 and ROR-gamma and known as a lymphoid
tissue inducer cell, which is critical for secondary and tertiary lymphoid tissue development and maintenance of CD4
T cell-dependent immune responses. Rheumatoid arthritis is chronic systemic inflammation disease that destroys
joint cartilages, bones and tissues. Some rheumatoid arthritis patients show that lymphocytes infiltrate to the
damaged joint and form lymphoid tissue-like tertiary lymphoid tissue structures in synovial tissue. The factors
involved in the formation of normal lymphoid tissues are expressed in abnormal lymphoid tissues of rheumatoid
arthritis synovial membrane such as CXCL13, CCL19, CCL21, CCR7, CXCR5, lymphotoxin-α/β, and IL-7.
2Rheumatology,
In this study, we compared percentages of immune cell populations of rheumatoid arthritis with osteoarthritis and
found that there are more macrophages in rheumatoid arthritis and more monocytes in osteoarthritis. In addition, we
found that the increased lymphocyte infiltration is correlated with an increase in lineage -CD4-CD127+ cells and
upregulation of OX40-ligand and TRANCE expression.
P2-011 SDF-1 INDUCES OSTEOCLASTOGENESIS IN RHEUMATOID ARTHRITIS BY UPREGULATING OF
RANKL EXPRESSION IN SYNOVIAL FIBROBLASTS AND CD4+ T CELLS
H.R. Kim1, S.H. Lee1, H. Kim2, K. Kim3, B.O.M.I. Kim4
1Rheumatology,
Konkuk University Hospital, Seoul, Korea
Soochunhyang University Hospital, Seoul, Korea
3Rheumatology, Catholic University of Korea, Seoul, Korea
4Rheumatology, Konkuk University of Korea, Seoul, Korea
Background and aims Stromal cell-derived factor(SDF)-1 is involved in bone destructive process in rheumatoid
arthritis(RA) and bony metastasis in malignancy, through inducing angiogenesis, producing matrix-degrading
enzymes and increasing survival and migration of osteoclasts. This study aimed to determine the mechanism of SDF1 on osteoclastogenesis in RA.
2Rheumatology,
Methods Synovial fibroblasts and CD4+ T cells were isolated from synovial tissues and peripheral blood of RA
patients. The expression of SDF-1 and RANKL was evaluated using confocal microscopy. After synovial fibroblasts
and CD4+ T cells were treated with rhSDF-1, the expression of RANKL mRNA was determined using real-time PCR.
Osteoclastogenesis was analyzed in culture of monocytes with SDF-1. Osteoclastogenesis was also determined after
monocytes were co-cultured with rhSDF-1-stimulated RA synovial fibroblasts and CD4+ T cells.
Results The expression of RANKL mRNA in RA synovial fibroblasts and CD4+ T cells was increased after SDF-1
stimulation. When RA synovial fibroblasts and CD4+ T cells were cultured with neutralizing antibody of TNF-a, the
SDF-1-induced RANKL expression decreased. When CD14+ monocytes were cultured with SDF-1 in the absence of
RANKL, the monocytes were differentiated into TRAP+ osteoclasts. Also, the monocytes were differentiated into
TRAP+ osteoclasts when they were co-cultured with SDF-1-prestimulated RA synovial fibroblasts or CD4+T cells.
Conclusion SDF-1 induced osteoclastogenesis by up-regulating RANKL expression in RA synovial fibroblasts and
CD4+T cells and this is mediated by TNF-a. The axis of SDF-1 and RANKL is a potential therapeutic target for bony
destructive process in RA.
P2-012 ANTI-ARTHRITIS EFFECTS OF (E)-2,4-BIS(P-HYDROXYPHENYL)-2-BUTENAL THROUGH INHIBITION
OF IKK-BETA? ACTIVITY
S. Nah1, S. Jeong2, S.O.R.A. Lee3, I. Kang4, H. Kim3
1Division
of Rheumatology department of internal medicine, Soonchunhyang University Cheonan Hospital, Cheonan,
Korea
2Department of psychiatry, Bright future clinic, Seoul, Korea
3Medical Research center, Soonchunhyang University College Of Medicine, Choenan, Korea
4Department of science, St. George's School, Vancouver British Columbia, Canada
Background and aims Maillard reaction products (MRPs) are known to have antioxidant, antimutagenic and
anticardiogenic activities. In the present study, we investigated whether (E)-2,4-bis(p-hydroxyphenyl)-2-butenal can
have anti-inflammatory activity and anti-arthritic activities through inhibition of NF-kB/IKK and STAT3 pathways in
cultured macrophage and synoviocytes, and collagen-induced arthritis (CIA) animal model.
Methods: Nitric oxide (NO) and prostaglandin E2 assay, electrophoretic mobility shift assay, luciferase assay,
Western blot, real-time PCR and pull-down assay were used for mechanism studies. Anti-inflammatory effect was
done in cultured RAW 264.7 cells and synoviocytes, and collagen-induced arthritis model was used for evaluation of
effects in vivo.
Results: (E)-2,4-bis(p-hydroxyphenyl)-2-butenal (2.5-10 mg/ml) inhibited LPS (1 mg/ml)-induced pro-inflammatory
responses through downregulating IkB kinase b (IKKb)/NF-kB and signal transducer and activator of transcription 3
(STAT3) pathways in RAW 264.7 cells and synoviocytes. (E)-2,4-bis(p-hydroxyphenyl)-2-butenal not only suppressed
the collagen (100 mg/0.1ml)-induced arthritic responses through inhibition of IKKb/NF-kB and STAT3 activities but
also reduced the extent of bone destruction and fibrosis in joint tissue. The population of white blood cells in blood
and NO generation in murine splenic T cells of collagen-induced arthritic mice were significantly reduced by (E)-2,4bis(p-hydroxyphenyl)-2-butenal. Sol-gel biochip analysis proved that (E)-2,4-bis(p-hydroxyphenyl)-2-butenal directly
binds to IKKb, and thus inhibit its activity. A docking experiment and pull-down assay indicate that IKKb might be a
potential target of (E)-2,4-bis(p-hydroxyphenyl)-2-butenal.
Conclusions: These findings indicated that (E)-2,4-bis(p-hydroxyphenyl)-2-butenal exerted its anti-inflammatory and
anti-arthritic effects through inhibition of IKKb/NF-kB activity via direct binding to IKKb, and that it could be a useful
agent for the treatment of rheumatoid arthritis
P2-013 EFFECTS OF ADIPOCYTOKINES TREATMENT ON FUNCTION OF ADIPOSE-DERIVED STEM CELLS
FROM RHEUMATOID ARTHRITIS AND OSTEOATHRITIS PATIENTS.
U. Skalska1, E. Kontny1, W. Maslinski1
1Department
of Pathophysiology and Immunology, Institute of Rheumatology, Warsaw, Poland
Introduction:
Adipose-derived stem cells (ASCs) have immunomodulatory properties and are intensively studied as a potential
therapeutical method to treat rheumatoid arthritis (RA). ASCs immunosuppresive function may be affected by
adipocytokines released by intra-articular adipose tissue. The aim of this study was to evaluate the influence of
high/middle molecular weight adiponectin isoform (HMW/MMW) and TNFα on immunomodulatory function of ASCs
from RA and osteoarthritis (OA) patients.
Materials and methods:
Infrapatellar fat pad was obtained from RA and OA patients. ASCs were isolated and cultured with/without
HMW/MMW (10μg/ml), TNFα (10ng/ml), IFNγ (10ng/ml). Expression of indoleamine 2,3-dioxygenase (IDO), heme
oxygenase 1 (HO-1) genes (Real Time PCR) and IL-6, IL-8, VEGF, TGFβ, IL-1Ra concentration in culture
supernatants (ELISA) were measured.
Results:
HO-1 was expressed in unstimulated ASCs and wasn`t influenced by studied factors. IFNγ upregulated strongly IDO
expression, TNFα had much weaker effect, whereas HMW/MMW caused only slight IDO gene transcription. OAASCs seem to have weaker expression of HO-1 and IDO genes. ASCs from RA and OA secrete spontaneously IL-6,
IL-8, TGFβ, VEGF, IL-1Ra. HMW/MMW significantly enhanced release of all cytokines by RA-ASCs, whereas TNFα
upregulated only IL-6 and IL-8. Comparing to RA-ASCs, secretion of IL-8 from OA-ASCs was much more enhanced
after HMW/MMW and TNFα stimulation (p<0,01).
Conclusion:
HMW/MMW and TNFα influence function of ASCs from RA and OA. ASCs from both diseases seem to be dissimilar
regarding their secretory activity and genes expression suggesting differences in their immunomodulatory properties.
Funding: grant no. 2011/01/N/NZ5/00932, National Science Center, Poland.
P2-014 A NOVEL APPROACH FOR TREATMENT OF RHEUMATOID ARTHRITIS
Z. Pan1, L. Chen2
1Medical
Microbiology and Immuniology, University of Toledo College of Medicine, Toledo, USA
University of Toledo College of Medicine, Toledo, USA
Background and aims: Rheumatoid arthritis (RA) is the most common inflammatory arthritis and a major cause of
various disabilities. Involvement of the NF-kB pathway in RA pathogenesis is well established. In the inflamed
synovium of rheumatoid arthritis patients, this is an overexpression of NF-kB, which is causes the transactivation of
various genes for pro-inflammatory cytokines, chemokines, adhesion molecules, MMPs, and inducible nitric oxide
(iNOS). Therefore, NF-kB plays a central role in the pathogenesis of RA by amplifying inflammation and causing joint
damage. Currently, the clinical drugs used to treat RA, consisting of NSAID and steroids, can help relieve pain and
reduce inflammation; however, the general effectiveness of these drugs have been unsatisfactory. Other therapeutic
modalities like TNFα inhibitors and IL-1 receptor antagonists are expensive and associated with serious side effects.
Therefore, the development of novel therapeutic strategies is urgently needed for the treatment of rheumatoid
arthritis. Results: Recently, we found that MKP-1, a MAP kinase phosphatase, negatively regulates the NF-kBdependent inflammatory response. In addition, we have found that Rolipram, which is already in late phase III clinical
trials for COPD, is a potent anti-arthritis agent by enhancing expression of MKP-1, a critical endogenous negative
regulator. Negative feedback regulation is an effective therapeutic strategy developed organically through evolution
that can combat an overactive inflammatory response without causing serious adverse effects. Conclusions: Our
studies will provide new insights into the molecular regulatory mechanisms underlying inflammation and provide a
novel therapeutic approach in a broad sense for treatment of autoimmune diseases.
2Medicine,
P2-015 PULMONARY MANIFESTATIONS IN RHEUMATOID ARTHRITIS
A. Zoto1, H. Hafizi2, T. Backa1, R. Osmenaj3, A. Harxhi4, Z. Ylli5
1Department
of Rheumatology, University Hospital Center “Mother Theresa”, Tirana, Albania
of Lung Diseases, University Hospital of Lung Diseases “Shefqet Ndroqi”, Tirana, Albania
3Department of Radiology, University Hospital Center “Mother Theresa”, Tirana, Albania
4Department of Infectious Disease, University Hospital Center “Mother Theresa”, Tirana, Albania
5Department of Immunology, University Hospital Center “Mother Theresa”, Tirana, Albania
Background and aims: Rheumatoid arthritis is a systemic autoimmune disease of connective tissue that
affects not only the articulations, but also various organs. Pulmonary involvement is one of the extraarticular manifestations of rheumatoid arthritis and is the cause of morbidity and mortality in patients with
rheumatoid arthritis.The aims of this study is the identification of lung injuries in patients with rheumatoid
arthritis and the impact of rheumatoid factor, anti cyclic citrullinated protein antibody and arthritis activity on
lung involvement in patients with rheumatoid arthritis.
2Department
Methods: This is a prospective study that analyzed 63 patients with rheumatoid arthritis. Physical
examination, clinical and immunological tests, pulmonary high-resolution computed tomography and
pulmonary function tests were performed for the patients. Rheumatoid arthritis activity was assessed
according to Disease Activity Score 28.
Results: Patients with positive rheumatoid factor are 39 (62%), anti cyclic citrullinated protein antibody
positive are 32 (51%) and active disease are 17 (27%). Interstitial lung disease is found in 22 (35%) patients,
pleural effusion are 3 (5%) patients and 7 (11%) patients bronchiectasi. 17 (27%) patients have restrictive
ventilator insufficiency, 4 (6%) patients have obstructive ventilator insufficiency. We found correlation
between pulmonary injuries with the severity of disease and the abnormality of immunological laboratory
tests.
Conclusion: Interstitial lung disease and restrictive ventilator insufficiency are injuries that occur more
frequently in patients with rheumatoid arthritis. Immunological alterations and the severity of the disease are
associated with pulmonary injuries.
P2-016 INHIBITORY EFFECT OF GLUCOSAMINE ANALOG NHAG ON THE PRODUCTION OF IL-6 AND MMP-2
AND MMP-9 IN RHEUMATOID ARTHRITIS DERMAL FIBROBLASTS
S.U. Simjee1, S.U.A. Shah1
1H.E.J.
Research Institute of Chemistry, International Center for Chemical and Biological Sciences University of
Karachi, Karachi, Pakistan
Rheumatoid arthritis (RA) fibroblasts participate in tissue destruction by producing interleukin-6, and
metalloproteinases which are involved in extracellular matrix degradation, and osteoclast-mediated bone resorption.
The present study was design to investigate the effects of NHAG (analog of glucosamine) on RA-DFBs. Primary
cultured DFBs of RA patient were cultured overnight in complete DMEM. Next day media was aspirated and DMEM
without FBS was added. The cells were pre-treated with NHAG (correspondent doses of 4, 20, 40, 60, 80, 100µM) for
4 hrs and then stimulated with TNF-α (5 ng/ml) for 24 hours at 37¡C with 5% CO2. After incubation, supernatants
were collected, centrifuged and used for determination of IL-6, MMP-2 and MMP-9. NHAG at the dose of 40 - 80 mM,
exhibited dose dependent inhibition of IL-6 production by RA-DFBs. The gel-zymography revealed that NHAG
markedly reduced the proactivated MMP-2 and MMP-9 in RA-DFBs at the dose of 80 and 100 mM. Further increase
in the NHAG dose was avoided because at higher concentration, the cell viability was compromised. In our previous
in vivo studies, NHAG has demonstrated a potent anti-arthritic activity. The present study further supports our earlier
findings indicating that some of the beneficial effect of NHAG seen in our rodent arthritic model might be due to the
inhibition of IL-6, and MMP-2 and MMP-9 as it was observed in in vitro study with RA-DFBs cells. This effect, coupled
with its multiple inhibitory effects on T lymphocyte functions, might account for the significant reduction in the rate of
disease progression in arthritic model treated with NHAG.
Poster Session: Session 2: Sjogren&apos;s syndrome
P2-017 CUTANEUS MANIFESTATION IN SJOGREN SYNDROME PATIENT TREATED WITH
HYDROXYCLOROQUINE
D. Mulya1, N. Sukmana2
1Internal
2Allergy
Medicine, Gadjah Mada University, Yogyakarta, Indonesia
and Clinical Immunology, University of Indonesia, Jakarta, Indonesia
Introduction:
Sjogren Syndrome is frequent autoimmune disorder (prevalence 0,5% - 2%) characterized by lymphocyte infiltration
of the salivary and lacrimal glands lead to dry eyes and mouth. Cutaneus skin manifestations in Sjogren Syndrome
are often underestimated, whilst 50% of Sjogren syndrome have cutaneus finding. This condition made patients with
Sjogren are wrongly diagnose nor had appropriate treatment earlier.
Case Presentation:
A 44 years old woman comes to our hospital with redness in her cheek since three years ago. She also felt pain in
her skin and getting worse every time she got exposure from the sun. Patient also complained for headache and
myalgia. From her physical examination we found malar rash and dry skin. The laboratory test showed positive Anti
Nuclear Antibody, dsDNA was 13,8 , SS A +++, and Anti RO S2 ++. We also checked for her schirmer test and the
result was 5 mm for right eye and 10 mm for left eye. Patients was treated with Hidroxychloroquine ,
Methylprednisolone 0,5 mg/Kg body weight, Vit D3, and Folic Acid. After one month therapy, patient showed
improvement in her skin manifestation and she no longer feel pain in her skin.
Conclusion:
We treated a 44 years old women with cutaneus manifestation as a symptom of Sjogren syndrome. Patients was
showed improvement while treated with Hidroxychloroquine , Methylprednisolone 0,5 mg/Kg body weight, Vit D3, and
Folic Acid.
P2-018 PRIMARY SJOGREN’S SYNDROME PRESENTING WITH PERICARDIAL AND PLEURAL EFFUSION
V. Selvan1, S. Xu1
1Medicine,
Khoo Teck Puat Hospital, Singapore, Singapore
Introduction
SjogrenÕs syndrome (SS) is a systemic autoimmune disease characterized by sicca symptoms. However, they are
rarely accompanied
by serositis such as pleuritis or pericarditis. We present a case report of a primary SjogrenÕs syndrome presenting
with pericardial and pleural effusion.
Case Report
The patient is a 50 year old Chinese female who was previously well. She presented with complaints of pleuritic chest
pain, cough, fever and shortness of breath for 7 days. Physical examination revealed decreased breaths sounds in
left lung base. She had a papulo-squamous lesion on her arm and leg. Chest xray showed bilateral pleural effusions
and & cardiomegaly. ECG showed small voltages with non-specific T wave changes. CT scan of thorax and abdomen
confirmed the presence of pericardial & pleural effusions. In addition there were mediastinal, axillary &
supraclavicular lymphadenopathy.
Blood investigations showed anemia of 9.8gm/dL, ESR 59 mm/1 st hr, CRP 66 mg/L. Pleural fluid analysis showed
exudative effusion which was negative for malignancy and infection. T-spot was negative. On direct questioning, the
patient admitted to a long history of dry eyes and dry mouth. Opthalmological review confirmed presence of
keratoconjunctivitis sicca. Tests for ANA (+), Anti Ro (+), Anti La (+), ds DNA (-), Rheumatoid Factor (+). Tests for
other ENA (-). She declined labial gland biopsy.
Primary SjogrenÕs syndrome was diagnosed based on the sicca symptoms, positive Ro and La but negative
dsDNA.The patient was treated with steroids and Hydroxychloroquine with improvement.
SjogrenÕs Syndrome should be considered as one of the connective tissue diseases potentially presenting with
pleural and pericardial effusion. This has been reported by other authors.
P2-019 THE IMPROVEMENT OF THE SURGICAL DENTAL CARE FOR PATIENTS WITH SJOEGREN'S DIS
EASE
E. Selifanova1, E. Selifanova1
1Stomatology,
Mental Health Research Center of RAMS, Moscow, Russia
Abstract:
There for, the purpose of this research is the improvement of organization of surgical dental care for patients with SD.
The evaluation of the nec essity of the surgical sanation was based on the following criteria: the ext ent of the tooth
crown destruction, tooth mobility, tooth retention and half -retention, tooth disposition, complicated with inflammation.
The surgical manipulations included tooth extraction, cystectomy, cystotomy, periosteototy and the operations on the
maxillary sinus. The therapy of the main disease and the focuses of odontogenic infection worsened the poor
regeneration of the operative site among the patients with SD, this manifestation is espe cially good represented in
cases f tooth extraction. The tooth sockets weren 't awfully filled with blood clot and postoperative healing required
nearly 3 weeks among 85% of patients. In 80% of cases postoperative alveolitis was presented, and the slowly
progressing osteomielitis was found among 35% of p atients. All the mentioned pathological processes required
additional common and local therapy. The poor condition of sutures on the 3d day of postopera tive period was
evaluated in 70% of cases. We recommend to make surgical sa nation of oral cavity among patients with SD after
cancel of cytostatic ther apy and intensifying of non-steroid therapy. Also we recommend to prescribe the
osteotrophic antibiotic therapy for 7 days and start it 2 before the ope ration and to prescribe antimicotic therapy in
order to prevent further complications.
P2-020 EXOCRINOPATHY MEDIATED BY ANTIMUSCARINIC AUTOANTIBODIES IN SJÖGREN’S SYNDROME
PATIENTS
K. Park1, M. Jin1, M. Park1, E. Namkoong1, S. Choi1
1Physiology,
School of Dentistry SNU, Seoul, Korea
Background and aims : Recently, functional autoantibodies against to muscarinic type 3 receptor (M3R) in
Sj*grenÕs syndrome (SS) has been reported. However, the pathological role of these autoantibodies in the
development of SS still remains to be elucidated. Methods: Purified IgGs were obtained from normal (control) and
primary SS patients' sera (pSS IgG). Anti-M3R autoantibodies in purified IgG were detected by M3R peptides using
ELISA assay kit. Surface expression and internalization of M3R and MHC I molecules were analyzed by
immunofluorescence confocal microscopy and biotinylation assay. Results: pSS IgG showed significantly higher
arbitrary units (AU) values in C2M3RP and C4M3RP than the other 3 peptide groups (one way ANOVA, P<0.05).
Surface expression of M3R, MHC I molecules but not β1AR was significantly decreased after incubation of human
SMG cells with pSS IgG. Internalization of colocalized M3R and MHC I molecules from the cell surface to the cell
interior during pSS IgG incubation was observed in hSMG cells and GFP·M3R transfected HSG cell line. Interaction
of M3R and MHC I molecules at the plasma membrane increased after incubation with pSS IgG but not control IgG.
The pSS IgG-induced M3R and MHC I internalization is prevented by the cholesterol-sequestering drug, filipin.
Conclusion: Cholesterol-rich microdomain of the plasma membrane containing M3R and MHC I molecules was
endocytosed by incubation of the cells with pSS IgG, which may partly explain the mechanism of exocrinopathy
shown in primary SS patients.
P2-021 ASSOCIATION BETWEEN HIGH CIRCULATING LEVELS OF INTERLEUKIN-18 AND DISEASE ACTIVITY
IN PRIMARY SJÖGREN'S SYNDROME
Y. Chen1, J.F. Zheng1, J.P. Yin1, R.L. Huang1, X. Gao2, X.H. Yu1, Z.G. Liu3, J. Chen4
1Medical
College Xiamen University, Xiamen University, Xiamen, China
Laboratory Department, Xiamen University Hospital, Xiamen, China
3Eye Institute and Affiliated Xiamen Eye Center of Xiamen University, Xiamen University, Xiamen, China
4Rheumatology Department, the First Afflicted Hospital of Xiamen University, Xiamen, China
Background and aims:
Interleukin-18 is a proinflammatory cytokine which plays an important role in infection, inflammation and
autoimmunity. Previous studies show elevated levels of IL-18 in primary Sj*gren's syndrome, an autoimmune disease
characterized by chronic inflammation in exocrine glands. We aimed to investigate the association between the levels
of IL-18, sIL-18R1 and IL-18BP and pSS and its subphenotypes.
Methods:Thirty-five well-characterized pSS patients and matched controls were recruited in this study. The serum
levels of IL-18, sIL-18R1 and IL-18BP were quantified using sandwich ELISA.
Results:Our results showed the levels of IL-18, but not IL-18BP or sIL-18R1 is higher in pSS patients than in controls
(119.73±100.06 pg/mL vs 65.03 ± 45.70 pg/mL, P<0.01). Furthermore, the levels of IL-18 in patients with active
disease are significantly higher than in patients with mildly active diseases (166.12±120.20 pg/mL vs 80.66
±57.86pg/mL, ,P<0.01).
Conclusions:To our knowledge, this is the first study that shows an association between IL-18 levels and the disease
activity of pSS. Our findings suggest that the circulating levels of IL-18 could be a biomarker indicating the disease
activity of pSS and IL-18 may be a potential therapeutic target.
Figure 1
2Clinical
Figure 2
group
IL-18(pg/mL)
IL-181(pg/mL) IL-18BP(ng/mL) Free IL-18(pg/mL)
119.73±100.06§
310.53±160.00 21.11±10.79
31.90±28.25§
pSS (n= 35)
28.57% (10/35) a* 11.43% (4/35)a 20% (7/35)a
20% (7/35)a*
65.03±45.70
262.89±123.18 20.33±4.44
16.83±12.31
5.71% (2/35)a
2.86% (1/35)a 5.71% (2/35)a
2.86% (1/35)a
Control (n=35 )
§Compared
with healthy controls, P<0.01,by Mann-Whitney Test,
a Compared
with healthy controls,P<0.05, by Fisher's exact Test.
Poster Session: Session 2: Skin and Autoimmunity
P2-022 INFLAMMATORY RESPONSES IN NONDERMATOMAL VITILIGO - A STUDY FROM SOUTH INDIA
Y. Ala1, M. Khalid Pasha1, V. Vijaya Lakshmi2, P. Jahan1
1Genetics,
University college of Science Osmania University, Hyderabad, India
and Molecular Biology, Lepra India-Blue Peter Public Health and Research Centre, Hyderabad, India
Background: Cytokines are the important mediators of immunity and disequilibrium in cytokine network may
paramount for autoimmune disease susceptibility and severity. Vitiligo is an acquired depigmenting disorder resulting
in the destruction of melanocytes in the epidermis affecting 1-4% worldwide with no prediction for sex or race. The
incidence of vitiligo is found to be 0.5 to 2.5% in India. The cause of vitiligo remains obscure, although the
autoimmune hypothesis has been now recognized as the most influential pathogenic mechanism. The present study
was conducted to determine the equilibrium between two important cytokines IFN-γ and IL-10 in nondermatomal
vitiligo and healthy individuals from South India.
2Immunology
Aim: To estimate the serum levels of IFN-γ and IL-10 in nondermatomal vitiligo patients and assess the ratio of these
proinflammatory and anti-inflammatory cytokines with respect to disease duration.
Methods: Estimation of IFN-γ and IL-10 levels in human serum from each of 60 nondermatomal vitiligo patients and
healthy controls using ELISA. Data analysis was performed using One-way ANOVA and t-test.
Results: Vitiligo patients showed increased levels of IFN- γ (11.7±5.2 vs 9.9±4.1 pg/ml) and decreased levels of IL-10
(9.4±5 vs 11.5±5.0 pg/ml) compared to controls. Ratio of IFN-γ to IL-10 was also significantly different in patients
(1.5±0.7 pg/ml) and controls (1.0±0.6 pg/ml), indicating association of proinflammation with nondermatomal vitiligo.
Further, significant correlation (p<0.05) was observed with disease duration.
Conclusion: Enhanced production of IFN-γ in patients may play a vital role in disease development suggesting Th1
mediated immune response in the pathogenesis of vitiligo.
P2-023 ASSOCIATION OF RAISED CAROTID INTIMA MEDIA THICKNESS WITH APOLIPOPROTEIN
B/APOLIPOPROTEIN A-I IN PATIENTS OF PSORIASIS.
K. Asha1, A. Singal2, S.B. Sharma1, A. Aggarwal3
1Department
of Biochemistry, University College of Medical Sciences, Delhi, India
of Dermatology, University College of Medical Sciences, Delhi, India
3Department of Medicine, University College of Medical Sciences, Delhi, India
Background and aims: Psoriasis patients are at increased risk of cardiovascular diseases (CVD), including
atherosclerosis. Carotid intima-media thickness (CIMT) has been recognized as a diagnostic tool to identify
premature atherosclerosis. Studies have shown that apolipoprotein measurement can help in prediction of CVD,
however association between apolipoprotein and CIMT need to be studied. The aim of this study was to measure
CIMT and estimate its relationship with apoB/apoA-I ratio in patients of psoriasis.
2Department
Methods: Serum lipid profile and apolipoprotein (apoA-I and apoB) were measured, in all hundred psoriasis patients
and age-sex matched healthy controls recruited for the study. Intima-media thickness and carotid plaques were
simultaneously measured by carotid sonography.
Results: The apoB/apoA-I ratio ( P < 0.001) and serum apoB ( P < 0.05) significantly correlated with raised CIMT in
patients. Conventional lipid profile and apoA-I levels were not correlated to IM thickness. By regression analysis we
found that apoB/apoA-I ratio was associated with CIMT.
Conclusions: The study shows that apoB/apoA-I ratio is associated with raised intima-media thickness and can be
considered as a predictor of sub-clinical atherosclerosis in patients of psoriasis.
P2-024 NEVIRAPINE INDUCED STEVENS JOHNSON SYNDROME
D. Madi1, C. Mahabala1, M.R. Pavan1
1Medicine,
Manipal university, mangalore, India
Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are rare immune mediated cutaneous
adverse reactions. The basic difference between SJS and TEN is the percentage of body surface area (BSA)
involved: <10% in SJS; >30% in TEN; 10 to 30% in SJS-TEN overlap. Drugs and infections commonly cause SJS.
Nevirapine a non-nucleoside reverse transcriptase inhibitor (NNRTI), is widely prescribed in India as a part of the
combination therapy to treat human immunodeficiency virus (HIV) infection. Nevirapine which is used to treat HIV can
also cause SJS. Patients infected with human immunodeficiency virus are at increased risk of developing
mucocutaneous drug reactions. We report a case of Nevirapine induced SJS.
A 47 year old gentleman,known case of HIV was started on Nevirapine based treatment as his CD4 cell count
dropped to 342 cells/mm³. Sixteen days later he presented with a mild skin rash on the trunk, conjunctivitis and fever.
Over the next few days the rashes became diffuse, bullous and progressed to involve the face, trunk and upper limbs
(Figure 1). Nevirapine was discontinued.He was treated with steroids and antibiotics.He improved completely over
next four weeks.
Although they are rare, these disorders have a high mortality rate. The most essential part of therapy is to discontinue
use of the suspected drug
.
P2-025 CLINICAL PROFILE OF CHILDHOOD AUTOIMMUNE BLISTERING DISEASES OVER FIVE YEARS
PERIOD FROM 2008 TO 2012
S.D. Shenoi1, R. Rao1
1Dermatology,
Kasturba Medical College, Manipal, India
BACKGROUND AND AIMS
Autoimmune blistering diseases ( AIBD ) in children are uncommon and are due to circulating antibodies to either
epidermal adhesion protein desmoglein or basement membrane zone components causing epidermal and
subepidermal blisters respectively.Amongst AIBD,most common is linear IgA disease (LAD ).Pemphigus vulgaris(PV)
pemphigus foliaceus( PF ),dermatitis herpetiformis ( DH )and bullous pemphigoid ( BP )rarely occur.The aim was to
study incidence and clinical profile of AIBD in children over five years period from 2008 to 2012.
METHODS:
31 children aged below 18 years with suspected AIBD underwent direct immunofluorescence ( DIF ).13 (7 girls, 6
boys ) confirmed cases of AIBD aged between 2 and 17 years were evaluated in detail.
RESULTS:
There were 7 cases of LAD,5 cases of BP and 1 of PV but none of DH or PF.Duration of disease ranged from 1month
to 3 years.All BP cases had oral involvement at presentation or during folowup while none of the LAD patients had
oral lesions during the course of the disease.The cutaneous lesions consisting of tense blisters were indistinguishable
in both BP and LAD.All LAD cases were treated with dapsone except one who developed anaemia to dapsone and
was given steroids. BP cases were treated with oral steroids and dapsone was added in one.All patients improved
with treatment except the patient with PV who expired following sepsis 10 days after admission.
CONCLUSION:
Direct immunofluorescence is the gold standard in the diagnosis of AIBD.Presence of oral lesions may be a marker of
childhood BP.
P2-026 TREATMENT OF ALOPECIA AREATA UNIVERSALIS WITH DIALYZABLE LEUKOCYTE EXTRACTS, A
REPORT OF 2 CASES.
T.A. Homberg1, M.C. Andaluz1, O. Pineda1, E. Cervantes1, M.A. Alonso1, M.C. Jimenez2, S.M. Pérez-Tapia1
1Immunology,
Instituto Politécnico Nacional, Mexico City, Mexico
Unit, I.A.P. Fundación Conde de Valenciana, Mexico City, Mexico
Alopecia areata is a T-lymphocyte mediated autoimmune disease of the hair follicle. Alopecia universalis is a severe
form that affects hair in the entire body. Initial presentation is often related to physical or emotional stress. No specific
treatment has proved successful on the long term. Although alopecia areata often presents spontaneous remission
over a few months, persistent cases cause patients to seek experimental and alternative treatment. We present two
cases of persistent alopecia areata universalis treated with dialyzable leucocyte extracts (DLE). Case 1 is a 4 year
old girl with alopecia universalis since age 2, not responding to previous topical treatment or corticosteroids. After 3
weeks of treatment with DLE there is growth of eyebrows, eyelashes and scalp hair, continuing for 5 more months.
Case 2 is a 38 year old female with alopecia universalis since age 34, with concomitant autoimmune hypothyroidism,
not responding to different treatments including immunosupressants. She presents 70% improvement after 4 months
of treatment with DLE.
2Research
Dialyzable leukocyte extracts are a human blood derived product composed of low molecular weight peptides ranging
from 1 to 12kDa in size. The proposed mechanism of action is induction of a Th1 immune response, which makes it
useful in the treatment of autoimmune disease. These cases represent a favorable response in severe and difficult to
treat cases of alopecia areata. Nonetheless, the effects after 5 months have not been evaluated and a larger simple
size is necessary to determine the effectiveness of DLE in this disease.
P2-027 EFFECTS OF ACUPOINT THERMAL STIMULATION ON ITCH IN THE MICE ATOPIC DERMATITIS
MODEL
Y.H. Chen1, W.C. Chen1, H.Y. Chen1, K.S. Tsai2
1Graduate
Institute of Integrated Medicine, China Medical University, Taichung, Taiwan
of Dermatology, China Medical University Hospital, Taichung, Taiwan
Background and aims: Chinese ancient records and several clinic trials showed that acupuncture make a significant
reduction of atopic dermatitis symptoms. We investigated the effects of different thermal stimulation at acupoint LI11
(Quchi) on scratching in mice elicited by subcutaneous administration of the pruritogenic agent (compound 48/80).
2Department
Methods: Ninety male ICR mice were divided into different groups: Normal saline s.c.; Compound 48/80 s.c.; and
Compound 48/80 s.c. with various thermal (5¡C, 15¡C, 25¡C, 35¡C and 45¡C) stimulation at L111. The mice were
acclimated individually in rectangular observation boxes for at least 2 hours. After different temperature stimulation (n
= 6/condition) was performed at LI11 acupoint in mice under isoflurane anesthesia for 20 minutes, pruritogens were
subcutaneous administrated on midline behind the neck of each mouse, and the scratch response was recorded. The
other two groups without temperature stimulation were subcutaneous administrated of pruritogens or normal saline
after anesthesia. Moreover, the cervical spinal cords (C5 - C7) were removed and fixed in paraformaldehyde solution
overnight at 4ºC. Six cervical sections from each animal were randomly selected and the number of c-fos positive
nuclei was counted under a light microscope.
Results: The application of lower temperature (15¡C) at the LI11 acupoint attenuated compound 48/80-induced
scratch. In addition, the application of lower temperature to the LI11 acupoint also decreased the number of c-fos
positive nuclei provoked by compound 48/80.
Conclusions: Lower temperature stimulation (15¡C) at the LI11 acupoint attenuated scratching behavior induced by
compound 48/80 in the mice atopic dermatitis model.
P2-028 MECHANISMS OF BLISTER INDUCTION BY AUTOANTIBODIES AGAINST THE EPIDERMAL
BASEMENT MEMBRANE
C. Sitaru1
1Dermatology,
Freiburg University, Freiburg, Germany
Collagens, laminins and other constituents of the epidermal basement membrane mediate a broad range of functions,
including tissue scaffolding, cell adhesion, cell migration, cancer, angiogenesis, tissue morphogenesis and tissue
repair. In addition to these roles, these components of the dermal-epidermal junction may also function as targets of
autoreactive T and B cells in autoimmune skin diseases. In a group of organ-specific autoimmune blistering diseases
the main autoantigens include collagen VII, the main constituent of the anchoring fibrils, the transmembrane
hemidesomosomal protein collagen XVII/BP180 and laminin 332. Autoimmunity to collagen XVII and laminin 332 is
associated with pemphigoid diseases, while collagen VII is the main autoantigen of epidermolysis bullosa acquisita.
Whereas autoreactive T cells are required for the initiation and modulation of the autoimmune response, tissue damage
resulting in subepidermal blistering is essentially mediated by autoantibodies. The mechanisms of blister formation are
multiple and include direct effects by steric hindrance and/or by triggering the transduction of a signal to the cell.
However, in addition to the binding to their target antigen, autoantibodies need to interact with factors of the innate
immune system, including the complement system and inflammatory cells, in order to induce full-blown disease.
Understanding the mechanisms by which autoantibodies specific to collagens and laminins disrupt the cell-matrix
adhesion should facilitate the development of more specific therapeutic strategies of autoimmune blistering diseases.
P2-029 CASE REPORTS SUCCESSFUL INTRAVENOUS IMMUNOGLOBULIN TREATMENT IN CHILDREN WITH
HYPER-IMMUNOGLOBULIN E SYNDROMES
B. Setiabudiawan1, R. Ghrahani1, G. Sapartini1, A. Sudjadi1
1Child
Health, Universitas Padjadjaran, Bandung, Indonesia
Hyper-immunoglobulin E syndromes (HIES) are very rare primary immunodeficiency which is characterized by clinical
triad of high serum level of Immunoglobulin E/IgE (>2000 IU/mL) and eosinophilia, recurring staphylococcal skin
abscess, and pneumonia with pneumatocele formation. GrimbacherÕs scoring system (NIH-HIES scoring system) is
used to diagnose HIES. Scores greater than 40% of the maximal score, based on age group, has a high significance
of HIES. The maximal NIH-HIES score for 3-6 year age group is 60 points. We reported a 5 year old girl who had
elevating total IgE serum level >2500 IU/mL, more than four lesions of skin abscess, pneumonia, which is proven by
3 times X-ray examination, serious skin infections (staphylococcal scaled skin syndrome), total eosinophil serum
1760 cells/µL, severe eczema and oral candidiasis. She had 41 points for total score, which was 60% of the total
score based on group of age, and we diagnosed HIES in our case based on this scoring systems. We used
intravenous immunoglobulin (IVIG), which may influence the IgE levels due to an increased immunoglobulin
catabolism or IgE neutralization via an anti-idiotype network. The patient was discharged with better clinical
presentation and laboratory findings two days after had IVIG therapy.
Poster Session: Session 2: Spondyloathritis
P2-030 THE USE OF PLASMAPHERESIS IN AN ANKYLOSING SPONDYLITIS PATIENT WITH SECONDARY
INFLIXIMAB FAILURE
S. Erdes1, O. Rumyantseva1, A. Bochkova1, S. Solovy?v2
1Spondyloarthritis,
Institute Rheumatology, Moscow, Russia
Care, Institute Rheumatology, Moscow, Russia
Background and aims. Increased titres of anti-infliximab antibodies can be one of the key causes of deteriorating
efficacy of the drug in the course of prolonged therapy. The goal of this observation is to assess the possibility of
overcoming secondary failure of infliximab in ankylosing spondylitis patients by using plasmapheresis.
2Intensive
Methods. One HLA-B2+ ASpts with peripheral arthritis and recurrent uveitis was monitored while receiving long-term
(>4y) treatment with infliximab (5mg/kg). The duration of disease is 12y, the duration of uveitis is 6y. After 2y of
treatment the duration of effect started to decrease reaching 4wks with relapses of arthritis and uveitis. During the 3y
of treatment, infliximab were administered every 6 weeks; uveitis recurred 4wks after each infusion, with high disease
activity: BASDAI-5.5; BASFI-6.4; nocturnal back pain-4(NRS); ESR-25mm/h; CRP-66mg/l. During the fourth year of
treatment, the patient received two plasmapheresis courses six months apart prior to scheduled infliximab infusions.
Results. A stable reduction of clinical and laboratory disease activity was achieved after a single course of
plasmapheresis (BASDAI-3.3; BASFI-5.5; NRS-3, ESR-5mm/h; CRP-7mg/l); arthritis was reduced, and intervals
between uveitis relapses increased (to 5wks after infusion). After second course of plasmapheresis: BASDAI-2.6;
BASFI-6.0; NRS-2; uveitis did not recur. A >20% improvement in the ASAS criteria was seen. Plasmapheresis had no
adverse effects.
Conclusions. Plasmapheresis allows to partially overcome secondary failure of infliximab. The immediate effects of
infliximab were enhanced with respect not only to spondylitis and peripheral arthritis, but also to uveitis.
P2-031 ASDAS AS A MARKER OF ANTI-DRUG ANTIBODY FORMATION IN ANKYLOSING SPONDYLITIS
Y. Kim1, S.C. Hong1, Y.J. Kim1, B.S. Koo1, C. Lee1, B. Yoo1, J.K. Ahn2, W.J. Seo3
1Internal
medicine division of Rheumatology, Asan Medical Center, Seoul, Korea
medicine division of Rheumatology, Kangbuk Samsung Hospital, Seoul, Korea
3Internal medicine division of Rheumatology, Seoul Veterans Hospital, Seoul, Korea
Background and aims
2Internal
Formation of antibodies to biologics has been regarded as main cause of treatment failure in rheumatoid arthritis and
ankylosing spondylitis (AS). However, assessment of anti-drug antibodies (ADA) is not available in routine practice till
now. Here, to find clinical markers predicting ADA formation in AS, we analyzed disease-associated parameters and
ADA presence.
Methods
In 20 AS patients initiating anti-TNFα therapy, blood was collected serially at baseline and 3-month after therapy and
serum-ADA levels were measured using ELISA. Disease-associated parameters including Ankylosing Spondylitis
Disease Activity Score (ASDAS), Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing
Spondylitis Functional Index (BASFI) and laboratory data including ESR/CRP were assessed.
Results
Among 20 patients (Adalimumab n=16, Infliximab n=4), ADA were detected in 4 (20%) patients (Adalimumab n=2,
Infliximab n=2). At baseline, there was no significant difference in laboratory data and clinical activity scores between
patients with and without ADA. After 3-month, only 1 of the 4 ADA-positive patients fulfilled clinically important
improvement (CII) criteria using ASDAS-CRP score (CII defined decrease ≥ 1.1 from baseline). However, all ADAnegative patients (n=16) achieved CII. (1/4 vs. 16/16, p=0.004). When assessed by ASDAS-ESR, CII achievement
was similar to that of ASDAS-CRP between ADA-positive and ADA-negative group. (1/4 vs. 15/16, p=0.013).
However, changes of BASDAI and BASFI score and number of patients who treated concomitantly with antirheumatic drugs or NSAIDs was not different in both groups.
Conclusions
In AS patients with anti-TNFα therapy, failure achieving CII using ASDAS at 3-month could suggest developing drug
immunogenicity.
P2-032 OBSERVATIONAL COHORT STUDY OF TNF-ALPHA GENOTYPE AND PHENOTYPE INCLUDING TNFINHIBITOR RESPONSE IN ANKYLOSING SPONDYLITIS
J. Nossent1, S. Johnsen2, G. Bakland3, J. Gran4
1Rheumatology,
Royal Darwin Hospital, Darwin, Australia
University Tromso, Tromso, Norway
3Rheumatology, University Hospital North Norway, Tromso, Norway
4Rheumatology, Oslo University Hospital, Oslo, Norway
Background The clinical efficacy of TNF inhibitors (TNFi) in patients with Ankylosing Spondylitis (AS) is apparent,
but the manner in which TNF-α contributes to disease pathogenesis remains unresolved. We investigated the
relationship between TNF-α gene promoter region polymorphism, serum TNF-α levels and clinical phenotype.
2Rheumatology,
Methods Cross sectional and longitudinal cohort study in TNFi naive AS patients from a regional disease registry.
Clinical and biological samples were collected during a research visit with genotyping for TNF-α -238 A/G and 308A/G performed by Taqman RT-PCR and TNF levels determined by sandwich ELISA. Subsequent TNFi treatment
during 8 years of follow-up was registered.
Results Three hundred and fifty five AS patients were genotyped and 14% presented the TNF-α *308 GA/AA
genotypes while 1% presented the TNF-α *238 GA/AA genotype. TNF-α *238 GA/AA genotype was associated with
later age of onset, lower ESR and higher hemoglobin, while the TNF-α *308 GA/AA genotype was associated with a
reduced risk of uveitis and better spinal function. Serum TNF-α level in AS patients (151 pg/ml) was lower than in
controls (263 pg/ml) as more AS patients had very low/undetectable serum TNF-α. (66 % vs. 25%, p<0.01). No effect
of TNF-α -308 / -238 or HLA-B27 genotype was seen on serum TNF-α. Genotypes were unrelated to the initiation of
TNFi or the number of TNFi agents used during eight years of follow-up, while TNFi therapy did not influence serum
TNF-α.
Conclusion The TNF-α -238 or -308 GA/AA genotypes in AS patients are associated with features of less severe
disease. Serum TNF-α is however undetectable in 2/3 of AS patients and not influenced by TNF-α promoter genotype
or TNFi therapy. These data indicate that the role of TNF-α in AS at local sites of inflammation supersedes its
systemic effect in AS patients.
Poster Session: Session 2: Systemic Lupus Erythematosus
P2-033 POTENTIAL IMMUNOPATHOLOGICAL ROLES OF THE NOVEL ANTI-INFLAMMATORY CYTOKINE
INTERLEUKIN-35 IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOUS
Z. Cai1, C.K. Wong1, L.S. Tam2
1Chemical
Pathology, The Chinese University of Hong Kong, Hong Kong, Hong Kong China
and Therapeutics, The Chinese University of Hong Kong, Hong Kong, Hong Kong China
Background and aims: IL-35, a heterodimers with subunits, IL-12A and Epstein-Barr virus induced 3, is a novel IL12 family immunosuppressive/anti-inflammatory cytokine. IL-35 is expressed by resting and activated regulatory T
cells (Tregs) by converting conventional T cells into IL-35-dependent induced Tregs. Similar to that of transforming
growth factor-b and IL-10, IL-35 is the major component of the suppressive repertoire, but temporally different from
them in the inhibition of inflammation. We hypothesize that IL-35 may play an important immunoregulatory roles in
systemic lupus erythematous (SLE).
2Medicine
Methodology: Plasma concentrations of IL-35 and soluble gp130 were measured using ELISA. Expression of IL-35
receptor components IL-12Rβ2 and gp130 on the CD4+ helper (Th) cells and CD19+ B cells, and the number of
CD4+CD25+CD127- Treg cells were quantitated by flow cytometry.
Results: Plasma IL-35 levels were significantly higher in active SLE patients than healthy controls (HC). Moreover,
percentages of Treg cells positively correlated with SLE disease activity index in active SLE patients, which were
significantly higher than inactive patients and HC. The expression of IL-12Rβ2 on the Th cells and B cells of inactive
and active SLE patients, respectively, was also significantly higher than HC.
Conclusion: The above results may imply the potential immunoregulatory roles of IL-35 receptors. Since IL-12Rβ2
played the key role in the signal transduction of the immunosuppressive mechanisms of IL-35 in SLE
immunopathogenesis, results of this cross-sectional study may furnish a biochemical basis for the development of
potential therapeutic target of IL-35 for the treatment of autoimmune inflammation.
P2-034 HIGH FREQUENCY OF CLINIC VISITS AND HOSPITAL ADMISSIONS FOR SLE AMONG FILIPINO
RHEUMATIC DISEASE PATIENTS IN A TERTIARY CARE CENTER
M. Galdones1
1Medicine,
University of Santo Tomas Hospital, Manila, Philippines
High Frequency of Clinic Visits and Hospital Admissions For SLE Among Filipino Rheumatic Disease
Patients In A Tertiary Care Center
Marian Esther G. GALDONES, Mhel Angelica CAMACHO, Sandra V. NAVARRA
yanski78@yahoo.com; 0922-8507872; 02-7499746
Rheumatology, Clinical Immunology and Osteoporosis, University of Santo Tomas Hospital, Manila, Philippines
Objective: To describe the frequency of clinic visits and hospital admissions among systemic lupus erythematosus
(SLE) patients relative to other rheumatic diseases at a tertiary care center in Manila, Philippines.
Methods: The 5-year patient census from January 2008 to December 2012 of University of Santo Tomas (UST)
Hospital was reviewed for the following information: primary rheumatic disease diagnoses and number of patients
under each diagnosis (each patient was counted only once for the entire 5 years). Patient encounters were
categorized as either clinic visits or hospital admissions.
Results: A total of 11,183 patients and 25,458 patient encounters were recorded from 2008 to 2012. Top 5 among
the primary rheumatic disease diagnoses were metabolic bone disease (MBD) 2,000 (18 %); soft tissue rheumatism
(STR) 1,895 (17%); osteoarthritis (OA) 1,877 (17 %); gout 1,619 (14%) and SLE 1331 (12%). Of the total 25,458
patient encounters, 21,732 (85%) were as out-patient clinic visits with SLE comprising the most number (6982 , 19%)
with a mean of 5.25 + 6.12 SD (range 1-42) visits/patient. SLE also comprised the highest number of hospital
admissions ( 1,664, 45 %), with a mean of 1.25 + 2.573 SD (range 1-23) admissions/ patient.
Conclusion: In this 5-year census of rheumatic diseases at the UST Hospital, SLE accounted for the highest
frequency of clinic visits and hospital admissions, reflecting the high burden of the disease.
Funding: Lupus-Inspired Advocacy (LUISA) Project of Rheumatology Educational Trust Fund Inc. (RETFI)
P2-035 SURVEY OF HEALTH CARE COSTS AMONG FILIPINOS WITH SYSTEMIC LUPUS ERYTHEMATOSUS
K. Geslani1, S.V. Navarra1
1Internal
Medicine Section of Rheumatology, University of Santo Tomas Hospital, Manila, Philippines
Objective: To survey the costs of health care in a cohort of Filipinos with systemic lupus erythematosus (SLE) seen
at a single tertiary care center.
Methods: This is a cross-sectional investigator-assisted survey conducted from March - June 2013 among SLE
patients aged > 18 years with at least 1 year duration of illness seen at the University of Santo Tomas Hospital,
Manila, Philippines. Survey instrument consisted of multiple choice and open-ended questions.
Results: Ninety-seven SLE patients (90 females) participated in the study. Average monthly cost of therapy was Php
5,793. Most patients (28.87%) had a combined monthly household income of Php8,000-15,000. Thirty-nine patients
alloted 11-25% of their monthly income for their medical condition. Allocation increased to as much as 700% with
infusion therapy, and 600% with hospitalization and hemodialysis. Twelve patients required hospitalization within the
past 12 months. 27 patients were on immunosuppressive agents for nephritis (11 on infusion therapy;16 on oral
therapy). 4 patients had end stage renal disease requiring dialysis.
Conclusions: We have described the wide spectrum of direct health care costs in a cohort of Filipino SLE patients,
illustrating the heavy economic burden of illness among those with renal involvement, on IV pulse therapy, and
frequent disease flares with complications requiring hospitalization. This preliminary data reiterates the need for early
and effective control of disease activity, in order to lessen the overall burden of illness on the patient, family, and
society.
[Currency conversion: US$ 1 = Php 41]
Funding: Lupus-Inspired Advocacy Project of Rheumatology Educational Trust Foundation Inc.
P2-036 HEART FAILURE IN A LUPUS PATIENT : FLARE OR VIRAL INFECTION ?
A.M. Frentescu1, I.C. Daha2, E.M. Tanasescu1, R.A. Ionescu1
1Internal
Medicine 3, Colentina Clinical University Hospital, Bucharest, Romania
Colentina Clinical University Hospital, Bucharest, Romania
Background and aim: Reporting of a particular heart involvement in a young lupus patient, emphasising the
complex possible etiology.
Method: We present the case of a 53-year-old woman, diagnosed with systemic lupus eritemathosus two months
prior to admission in our department, who than seeked medical care for disponea, that appeared during physical
exercise, bilateral oedema of the anterior calf region and tiredness. It is noteworthy that, six months before, the
patient had a pericarditis episode, without any damage of the cardiac function.
Results: Laboratory tests revelaed active lupus (leukopenia, marked proteinuria, high ESR and CRP) and cardiac
failure (LVEF=40%, diastolic disfunction of the left ventricule, hipokinesia of the left ventricule,
NTproBNP=3451pg/mL). Pulse-therapy with methylprednisolone, 500 mg/day, for three consecutive days, was
initiated with significant clinical improvement in aproximately 36 hours
2Cardiology,
Conclusion: At this patient, the etiology of the cardiac failure raises a number of complex problems as it may be due
to the lupus disease (lupic miocarditis, coronary vasculitis and/or early atherosclerosis ) or to an episode of viral
miocarditis that may have even triggered the lupus flare (as CRP levels were raised, even though, lupus disease
presents with normal levels). Considering the rapid favorable evolution of the symptoms, it is believed that the cardiac
disfuntion is part of the lupus disease. Unfortunately, neither viral tests, nor endomyocardial biopsy were performed.
P2-037 RENAL ARTERY THROMBOSIS IN ANTIPHOSPHOLIPID ANTIBODIES-NEGATIVE PEDIATRIC LUPUS
PATIENT
H. Lee1, J.Y. Choe1, S.K. Kim1, S.H. Park1, K.Y. Park2, J.H. Kim2
1Rheumatology,
Daegu Catholic university Hospital, Daegu, Korea
and Autoimmunity Research Center, Daegu Catholic university Hospital, Daegu, Korea
Background: Systemic lupus erythematosus (SLE) is an autoimmune disease that affects multiple organ systems with
immune complexes and a large array of autoantibodies. Thrombosis has been reported in about 10%-26% of SLE
patients and antiphospholipid antibodies(aPL) are important risk factors for thrombosis. Renal artery thrombosis has
rarely been reported in young patients, and no pediatric cases about aPL-negative SLE patients with renal artery
thrombosis have been reported in Korea.
2Arthritis
Case : A 12-year-old female patient presented to the hospital with a 3-day history of nausea, vomiting, and right flank
pain. She was diagnosed with SLE with lupus nephritis two years ago and she has been treated with a corticosteroid,
mycophenolate mofetil and hydroxychloroquine. Right costovertebral angle tenderness was revealed on physical
examination. Initial laboratory investigations revealed that lupus anticoagulant antibody, anti-beta2 glycoprotein-I
antibody, anti-cardiolipin Ab lgG and IgM were negative. Abdominal computed tomography showed renal artery
thrombosis and renal infarction. Subcutaneous low molecular weighted heparin was started and concurrently,
warfarin was started and has been continued for 3 months. After 3 months, 2.2cm sized heterogenous hypodense
lesion at the upper pole of right kidney was not seen and it lefted small atrophic changes in the abdominal computed
tomography. She is doing well by maintaining oral anticoagulant and followed up regularly through outpatient clinic
visits.
Conclusion : Renal infarction should be suspected in cases of unexplained sudden flank or upper abdominal pain,
with or without fever, nausea, and vomiting even if antiphospholipid antibody is negative.
P2-038 CONTRIBUTION OF T-INDEPENDENT AND T-DEPENDENT MECHANISMS, MONOCYTES AND NK
CELLS IN SYSTEMIC LUPUS ERYTHEMATOSUS
L. Lee1, C. Zhu1, S. Adelstein2, B. Fazekas de St Groth1
1T
Cell Biology, Centenary Institute, Sydney, Australia
of Clinical Immunology, Royal Prince Alfred Hospital, Sydney, Australia
Animal models demonstrate two independent mechanisms leading to systemic lupus erythematosus (SLE):
overexpression of B-Cell-Activating-Factor (BAFF) in a T-independent manner, and abnormal activation of follicularT-helper (Tfh) cells with normal BAFF levels. In SLE patients, it has been reported that disease severity correlates
with increased serum BAFF levels, increased circulating Tfh cells and several B cell abnormalities. Monocyte
involvement in the pathogenesis of SLE has been proposed to act through enhanced recognition of immune
complexes containing TLR-binding self-antigens. Total numbers of NK cells are decreased in SLE patients, but NK
subset distribution is similar to healthy controls. It remains unclear whether these abnormalities co-exist within
patients, or if human SLE can be divided into subcategories depending on the immune process driving the disease.
2Department
Flow cytometric analysis of PBMCs in 30 SLE samples indicated that frequency of CD27 -IgD- memory B cells was
increased in SLE patients and inversely correlated with disease severity. While some SLE patients had a greatly
increased population of transitional B cells, other SLE and control samples had almost none. Neither patients nor
controls had clearly identifiable populations of circulating Tfh cells. Frequency of CD56 briCD16- NK cells was
significantly higher in SLE patients than controls, while frequency of CD56dimCD16+ NK cells was significantly
decreased in SLE patients compared with healthy controls.
Measurement of BAFF levels is currently being performed. This study will be extended to a larger cohort of patients.
Our results will indicate the relative contributions of T-dependent and T-independent mechanisms, monocytes and
NK cells in human SLE.
P2-039 COMPARISON OF MYCOPHENOLATE MOFETIL AND INTRAVENOUS CYCLOPHOSPHAMIDE AS
INDUCTION THERAPY IN KOREAN PATIENTS WITH LUPUS NEPHRITIS
S. Lee1, D. Park1, K. Lee1, L. Wen1, J. Lee1, J. Kang1
1Rheumatology,
Chonnam National University Hospital, Gwangju, Korea
Background: To compare the efficacy of mycophenolate mofetil (MMF) and intravenous cyclophosphamide (IVC)
pulses as induction treatment for lupus nephritis (LN) in ethnically homogeneous Korean patients.
Methods: This study enrolled 49 LN patients with available kidney biopsy specimens. The renal biopsy specimens
were reclassified according to the ISN-RPS classification, by a renal pathologist blinded to the previous classification.
The renal outcome, i.e., complete response (CR), partial response (PR), and non-response (NR), after 6 and 12
months was defined according to the ACR 2006 response criteria for proliferative and membranous renal disease in
clinical trials.
Results: Of the 49 patients, 28 (57.1%) were treated with IVC and 21 (42.9%) with MMF, both in combination with
prednisolone. The baseline characteristics of the two groups were comparable, except that the IVC-treated patients
had lower platelet counts, lower C3 levels, and higher activity scores compared to the MMF-treated patients. CR was
seen in 42.9% receiving MMF and 50.0% receiving IVC after 6 months treatment (p= 0.450) and in 52.4% in the MMF
group and 46.4% in the IVC group at 1 year (p= 0.745). The number of patients achieving PR and NR did not differ
significantly at 6 and 12 months between the treatment groups.
Conclusion: These findings suggest that the efficacy of oral MMF at 1 year does not differ from that of IVC in
induction treatment of LN in ethnically homogeneous Korean patients. MMF may be considered first-line induction
therapy for treating LN in these patients.
P2-040 CHEMOKINE LEVELS AND AUTOANTIBODIES PRODUCTION IN SYSTEMIC LUPUS
ERYTHEMATOSUS
B. Leishangthem1, A. Wanchu2, A. Sharma3, A. Bhatnagar1
1Biochemistry,
Panjab University Chandigarh, Chandigarh, India
of Arthritis and Rheumatic Diseases-OP09, Oregon Health and Science University, Portland, USA
3Department of Internal Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India
Background and Aim: Systemic lupus Erythematosus (SLE) is chronic inflammatory and complex autoimmune
disease. It affects various parts of our body because of autoantigens. RANTES, chemokine and its receptor CCR5
have been associated in the pathogenesis of SLE. So a study was designed to evaluate association of anti-ds DNA,
anti- Ro, RANTES and CCR5 with disease activity.
2Division
Methods: For this study, blood was collected (after informed consent) from 80 SLE patients diagnosed and
undergoing treatment in PGIMER along with 80 healthy controls. Levels of anti *ds DNA, anti-Ro levels and RANTES
were measured using ELISA kits. Whereas the expression of CCR5 was evaluated by using FITC tagged monoclonal
antibody in flow cytometer. Disease activity score was calculated by Systemic Lupus Erythematosus disease Activity
(SLEDAI).
Results: The levels of RANTES and CCR5 expression were found to be higher in patients 1840.48±739.42pg/ml vs
835.44±70.48pg/ml; P<0.0001 and 26.49±.16% vs 24.72±3.02%; P<0.05 respectively as compared to controls.
Autoantibodies levels were also found to be higher in patients. SLEDAI score was found to be negatively correlated
with elevated anti-dsDNA (p<0.05). But a moderately significant negative correlation as compared to controls was
found in borderline patients for anti-Ro autoantibody (p<0.01). The levels of RANTES and CCR5 were also higher in
case of patients than controls. But there was no significant correlation of RANTES and CCR5 with disease activity.
Conclusion: The study shows an association of auto-antibodies, RANTES and chemokines levels with SLE. Later
studies should emphasize on the anti-SSA during the diagnosis of disease.
P2-041 DETECTION OF SM ANTIBODIES USING SMD3 SYNTHETIC PEPTIDE
W. Papisch1
1International
Sales and Marketing, Phadia GmbH (part of Thermofisher Scientific), Freiburg, Germany
Background
Antibodies against Sm are highly specific for SLE. They are found in 5-30% of SSL patients - more frequent in
Asian/African Americans (30-40%) compared to Latin Americans/Caucasians. The Sm antigen consists of different
proteins (SmD, SmBBÕ and others). SLE-specific Sm antibodies are those directed against SmD proteins and thus
these must be the ones referred to. ELISA based techniques are highly dependent on the quality of the antigen. A
new approach is the use of well-defined and highly sensitive and specific SmD3 peptide as antigen.
Objective
Analyse the clinical performance of a synthetic SmD3 peptid in a fully automted system compared to an Elisa with
conventional Sm.
Material and Methods
Sm antibodies were measured in a cohort of 350 samples using EliA SmD P performed on the fully automated Phadia
250 instrument. Clinical performance data such as clinical sensitivity and specificity as well as predictive values and
likelihood ratios were calculated.
Results
Clinical performance data obtained by the various assays are outlined in detail below.
EliA SmDP performs better than the compared Elisa with conventionally purified Sm preparation resulting in a cleary
higher positive likelihood ratio supporting the high clinical value. The comparatively low sensitivity of 23% is due to
the prevalence of the marker in SLE in the patients with Caucasian background.
Conclusion
o
o
o
Results obtained by SmDP petide are highly sensitive and specific
Data are superior to conventionally purified Sm
The approach by using synthitc Sm Peptides may overcome the issues apparent when using
conventionally purified Sm
P2-042 ORGAN INVOLVEMENT AND TREATMENT EFFECT OF TUBERCULOSIS ON STEROID REQUIREMENT
AMONG FILIPINO PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS
J. Patricio1, M.T. Salvador1, S.V. Navarra1
1Department
of Medicine Section of Rheumatology, University of Santo Tomas Hospital, Manila, Philippines
INTRODUCTION. Tuberculosis (TB) is a leading cause of morbidity and mortality in SLE particularly in endemic
countries. We describe the organ involvement and outcomes of TB infection among Filipino SLE patients, and
analyzed the effect of adequate TB treatment on corticosteroid requirement.
METHODS.Retrospective review of medical records of SLE patients seen at the Rheumatology Clinics of USTH who
completed a minimum 6-month TB therapy was done. Average prednisone (mg/day), disease activity scores and
selected laboratory parameters were recorded before, during, and upon completion of therapy.
RESULTS. There were 153 episodes of TB infection in 122 SLE patients (112 females, 92%). Average age was
29.30+12.97 (range 7-67) at SLE diagnosis, with duration of 88.5+64.2 months (range 1-276) to TB diagnosis.
Pulmonary TB (PTB) involvement was seen in 62%. 43% had extra-pulmonary TB (EPTB), 11.76% having solely
EPTB while 13.73% were disseminated TB. EPTB included meningitis (26%), soft tissue (22%), arthritis (20.75%),
genitourinary, gastrointestinal, spine and pericarditis. Average daily prednisone (mg/day) decreased from
16.45+12.58 before to 10.85+7.93 during (p<0.05), and 7.13+5.36 after TB therapy (P<0.05). Overall SLEDAI scores
decreased after TB therapy. Hemoglobin (g/L) increased from 104.34+17.20 to 113.62+11.98 (p<0.05) and
sedimentation rate decreased from 76.34+32.69to 26.96+15.43 post-TB therapy.
CONCLUSION. Appropriate TB therapy positively impacts SLE by decreasing steroid requirement, potentially
decreasing overall disease activity. Expected improvement in anemia and ESR levels suggest possible confounding
effect of TB infection in assessment of their disease activity.
Funding: Lupus-Inspired Advocacy( LUISA) of Rheumatology Educational Trust Fund Inc (RETFI)
P2-043 GASTROINTESTINAL INVOLVEMENT IN SYSTEMIC LUPUS ERYTHEMATOSUS: A CASE SERIES
J.P. Patricio1, J.D. Ngo1, S.V. Navarra1
1Department
of Medicine Section of Rheumatology, University of Santo Tomas Hospital, Manila, Philippines
Introduction: Systemic lupus erythematosus (SLE) presents with protean clinical manifestations, with infrequent
involvement of the gastrointestinal (GI) tract. This reports 3 Filipino patients whose lupus flare consisted primarily of
GI involvement.
Case1: 34 year old diagnosed SLE of 14 years, stable on prednisone 5mg/day and hydroxychloroquine presented
with vomiting, diarrhea, abdominal pain, proteinuria and hypocomplementemia. Radiograph showed focal segmental
ileus and dilated small bowel loops. Abdominal ultrasound revealed diffuse hepatic parenchymal changes, ascites
and incidental pleural effusion. Her symptoms dramatically improved on high dose steroids.
Case2: 44 year old SLE of 19 years duration maintained on prednisone 15mg/day and hydroxychloroquine for
thrombocytopenia developed recurrent epigastric pain and vomiting. Esophagogastroduodenoscopy showed acute
gastric mucosal erosions. Her symptoms were unresponsive to GI medications. High dose steroids provide almost
immediate relief and there were no recurrences while tapering steroids. Cyclophosphamide therapy was eventually
started for persistent thrombocytopenia.
Case3: 24 year old recently diagnosed SLE with mucocutaneous lesions, positive ANA and hypocomplementemia
presented with worsening epigastric pain and vomiting unresponsive to GI medications. Radiograph and CT scan
showed ileus with pneumo-peritoneum suggestive of ruptured viscus. Exploratory laparotomy revealed gangrenous
terminal ileum; resection with colostomy was performed. High dose steroids were simultaneously started. She was
discharged improved on tapering steroids.
Conclusion. We report 3 patients with GI involvement associated with active SLE, all of whom improved with high
dose steroids. Early recognition of this infrequent involvement in SLE improves the prognosis and lessens overall
morbidity.
Funding: Lupus-Inspired Advocacy (LUISA) of Rheumatology Educational Trust Fund Inc.
P2-044 I. EVALUATION OF THE EFFECTS OF APPLICATION OF PULSE THERAPIES IN PATIENTS WITH
NEUROLOGICAL MANIFESTATIONS OF SYSTEMIC LUPUS ERYTHEMATOSUS
V. Reshkova1, R. Rashkov1, S. Monov1, D. Kalinova1, T. Yoneva1
1Clinic
of rheumatology, University hospital 'St. Ivan Rilski", Sofia, Bulgaria
The Systemic Lupus Erythematosus (SLE) is manifesting itself with different clinic symptoms and immune
phenomena. The affectation of the Central Nervous System (CNS) manifests severe effects and it is difficult to treat
revealing unfavorable prognosis factors as well.
The purpose of the present clinical study was to evaluate the effects of the long term application of
Methylprednisolone and Cyclophosphamide Pulse Therapy in patients with manifested Central Nervous System
Lupus in their clinical picture.
In the Clinic of Rheumatology in Sofia took place a clinical treatment of 35 female for 12-18 months with intravenous
Methylprednisolone and Cyclophosphamide Pulse Therapy. The clinical status of the patients was evaluated
applying the SLEDAI, an immunological analysis and MRI of CEREBELUM.
During the studied period was observed the reduction and even the disappearance of symptoms.
1. At the beginning 18 patients (52,43%) were with epileptic convulsions, meantime with 12 patients (66,7%) was
observed a reduction and even disappearance of the epileptic convulsions.
2. The organic syndrome was diagnosticated with 18 patients (51,43%) while 9 of the patients (50%) ameliorated
their clinical status.
3. The Lupus Headache was the most difficult one to treat.
The efficiency of Pulse Therapy is evident influencing successfully the severe symptoms in the CNS Syndromes of
SLE, improving and restoring the pertinent functions and increasing the patientsÕ quality of life.
The of is evident influencing successfully the severe symptoms in the CNS Syndromes of SLE, improving and
restoring the pertinent functions and increasing the patientsÕ quality of life.
P2-045 LUPUS GENETICS: WHAT GWAS HAS MISSED?
W. Yang1, Y.L. Lau1
1Peadiatrics
and Adolescent Medicine, The University Of Hong Kong, hong kong, Hong Kong China
Systemic lupus erythematosus is a prototype autoimmune disease with a strong genetic component. Asians
have higher disease prevalence and lupus nephritis than European populations. This is consistent with our
findings from meta-analysis of GWAS on two Chinese populations. Overall, around half of the susceptibility
loci identified from Asian studies so far are likely to be different between Asians and European populations.
This raises the question that for future risk prediction and personalized management, the analysis has to be
based on studies from the relevant populations. ITGAM is a good example for subphenotype stratification
and population differences, with its different allele frequencies between Asians and European populations
and association with lupus nephritis. The HLA region showed most prominent association with many
autoimmune diseases. Population differences and independent contributors are also prominent for this
region. Thus detailed understanding of this region is important in further explaining the missing heritability,
subphenotype stratification, and personalized management of the disease. Typing the HLA alleles is
somewhat difficult; thus we developed a method for HLA typing using next generation sequencing with
flexibility, reasonable throughput and very low cost. This method is not only suitable for HLA typing for
lupus studies, it can also be used for genetic screening of other genetic disorders and for population
screening.
P2-046 INCREASED APOPTOSIS AND ABERRANT APOPTOSIS SIGNALING PATHWAYS OF
CD4+CD25+FOXP3+ REGULATORY T CELLS IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS
L. Na1
1Dermatology,
Southwest Hospital Third Military Medical University, Chongqing, China
Background and aims:Decreased cell numbers and suppressive defects of naturally occurring CD4 +CD25+FOXP3+
regulatory T cells (Tregs) play an important role in the breakdown of SLE immunetolenrance. We have peviously
observed significantly increased apoptosis of peripheral blood CD4 + T cells in SLE patients. Our objective here was
to detect the apoptosis of Tregs in SLE patients to see if it could contribute to reduced suppressive activity of Tregs,
and further elucidate the genes and signaling pathways which trigger the apoptosis in these cells. Methods: The
apoptosis of Tregs was evaluated in SLE patients and normal controls(NCs) by FACS. The suppressive activity of
Tregs was measured by coculture with CD4+CD25-CD127dim/- T cells. The relationship of abnormal Tregs apoptosis
with clinical parameters was analyzed by Pearson correlation analysis. Gene expression profiles of unstimulated
Tregs from active SLE patients and NCs were generated by microarray analysis. Differential genes expression were
verified by real time-PCR. Results: We found that the Tregs from SLE patients showed a significant elevation in
apoptosis and reduction in suppressive capacity compared to NCs. The increased apoptosis of Tregs was negatively
correlated with the total number of Tregs and positively correlated with disease activities. Gene expression profiles of
unstimulated Tregs from recent-onset SLE subjects reveal a cellular response that could make the cells sensitive to
apoptosis, partially due to the stress responses and cytokine stimulation. Conclusions: This global picture of
pathway-specific expression signatures is a step further into dissecting Tregs defects in the pathogenesis of SLE.
P2-047 HYPERHOMOCYSTEINEMIA ASSOCIATED WITH INCREASED BONE RESORPTION AND DECREASED
BONE FORMATION MARKERS IN FEMALE PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS
K. Handono1, T. Luthfia Sari2, A. Amelia Adam2, M. Zulbaidah Artamin1
1Clinical
Pathology Faculty of Medicine, Brawijaya University, Malang, Indonesia
of Biomedical Science Faculty of Medicine, Brawijaya University, Malang, Indonesia
Background. Premature osteoporosis is one of the long term complications of Systemic Lupus Erythematosus
(SLE). Recent studies showed that increased level of homocysteine was detected in SLE patients and it was
associated with deterioration of bone health. The aim of this study was to determine the association between
homocysteine level and biochemical parameters of bone turn over in SLE patients.
2Laboratory
Subyects and Methods. Thirty-nine female patients who fulfilled ARA criteria for the classification of SLE under the
age of 50 years were studied. All patients were moderately active ( SLEDAI >5). Bone turn over were assessed using
βCTx ( bone degradation) and osteocalcin ( bone formation ). Laboratory parameters including serum
homocysteine??, folic acid, vitamin B6, vitamin B12, βCTx, and osteocalcin were measured by ELISA method. Blood
samples were taken after 10 hours fasting.
Result.This study found that 71.8% of SLE patients had hyperhomocysteinemia. High homocysteine level was
associated with increased levels of βCTx (p = 0.000, R = 0.943) and decreased levels of osteocalcin (p = 0.000, R =
0.771). Serum levels of folic acid, vitamin B6, and vitamin B12 tend to be low in SLE patients. A high incidence of
hyperhomocysteinemia was found in premenopausal patients with SLE which affect bone metabolism. Bone
diminution in SLE seems to be attributable both increased bone resorption and decreased bone formation.
Keywords : SLE, homocysteine, folic acid, B6, B12, βCTx, osteocalcin, osteoporosis
P2-048 CIRCULATING MICROPARTICLE DOUBLE-STRANDED DEOXYRIBONUCLEIC ACID IN SYSTEMIC
LUPUS ERYTHEMATOSUS – ENUMERATION AND CLINICAL DISEASE ACTIVITY CORRELATION
S.H. Tay1, A. Lateef1, D.R. Koh2, Y.C. Lim3, I. Ng1, L.H.K. Lim2
1Medicine,
National University Health System, Singapore, Singapore
National University of Singapore, Singapore, Singapore
3Pathology, National University of Singapore, Singapore, Singapore
Background: Microparticles (MP) have been postulated to be autoadjuvants in the pathogenesis of systemic lupus
erythmatosus (SLE). Herein, we aim to establish the presence of extracellular DNA in the form of circulating MP DNA
and attempt to correlate MP DNA levels with disease activity.
2Physiology,
Methods: MPs from plasma from SLE patients and healthy controls were pelleted using high-speed centrifuge and
analyzed using flow cytometry. PicoGreen doubled stranded DNA (dsDNA) quantitation reagent was used for
staining. The sensitivity of MPs to detergent Triton X-100 was assessed. Sensitivity to detergent lysis allowed for
differentiation from MP dsDNA from unassociated (MP-free) dsDNA. Enumerated MP dsDNA were purified from the
unassociated dsDNA using a membrane ultrafiltration spin column.
Results: MP DNA of 14 SLE patients and 8 healthy controls were quantified. The median MP DNA/uL of plasma for
SLE patients was significantly higher than healthy controls 2460.28 (Q1; Q3 1010.91; 3416.26) vs. 403.73 (Q1; Q3
222.26; 1801.88) (p=0.020). There was significant correlation of MP DNA concentration with anti-dsDNA
immunoglobulin G (IgG) serum titres in SLE patients (r=0.818, p=0.002). MP DNA concentration correlated negatively
with complement 3 levels (r=-0.501, p=0.034). The purity of MP DNA was increased from 9.22% to 83.20% following
concentration with the ultrafiltration column.
Conclusion: We have demonstrated the presence of MP DNA in plasma of SLE patients and healthy controls.
However, the level of MP DNA in SLE patients was significantly higher than healthy controls. The level of MP DNA
also correlated strongly with titers of anti-dsDNA IgG in SLE patients.
P2-049 AUGMENTED ANTIBODY PRODUCTION IS CONSISTENT WITH INCREASED ADHESION BETWEEN
ACTIVATED CD4 T AND B CELL INTERACTION IN SYSTEM LUPUS ERYTHEMATOSUS
Y. Wang1, Z.C. Liu1, S.J. Wang1
1Department
of Immunology and Microbiology Shanghai Jiaotong University School of Medicine, Shanghai Institute of
Immunology, Shanghai, China
Background and aims: Molecular dissection of how T cell activation promoting B cell functionality will provide
important clues to the pathogenesis of autoantibody-involved SLE pathogenesis.
Methods: Using CD4 T cells either from SLE patients or activated in vitro by anti-CD3/CD28 antibodies, we have
detected the adhesion between CD4 T and B cells by using a micropipette adhesion frequency assay. Activated
CD4+T were further co-cultured with na•ve B cells in vitro. IgG and IgM levels in the supernatant were detected by
ELISA.
Results: The adhesion between activated CD4 T cells from SLE patients and B cell was much stronger than that of
normal CD4T cells. With the addition of anti-ICAM-1 or anti-ICOS blockade antibodies separately, the adhesion
frequency decreased. Consistent to the adhesion, IgG and IgM levels in the supernatant increased dramatically from
day 4 whereas there was very little IgG and IgM detectable with the co-culture of non-activated normal T cells and B
cells. The addition of anti-ICAM-1 antibodies largely blocked the production of IgG and IgM. Augmented IgG
production by B cells was also observed after co-culture with CD4 T cells from SLE patients.
Conclusions: Our results aforementioned demonstrated that CD4 T cells from SLE enhanced B cell-mediated
antibody production, probably initiated through increased adhesion between T and B cells. ICAM-1 played important
roles on both processes. This might lay the molecular foundation to study the roles of T/B cell interaction on the
pathogenesis of antibody-driven SLE as well as facilitate the screening of optimized therapeutic targets in SLE.
P2-050 A PREGNANT WOMAN WITH SEVERE LUPUS FLARE
D. Fetarayani1, A. Awalia1
1Internal
Medicine, Medical Faculty of Airlangga University, Surabaya, Indonesia
BACKGROUND
Pregnancy in an SLE patient is associated with increased risk of adverse maternal and fetal outcomes. Lupus
disease activity may flare during pregnancy or in the immediate postpartum period. Flares occur in up to 61-67% of
patients if the disease was active at conception.
CASE
A 24 year-old woman, 7thmonth of pregnancy, came to ER due to generalized edema since 3 months ago and got
worse in the last 2 days. She complained body weakness, loss of appetite, nausea, hair loss, and joint pain since 1
week before. She got SLE for 4 years but without treatment for the last 1 year. Examinations showed anemia, malar
rash, and both legs edema. Hemoglobin level 7.1g/dL, ESR 89mm/hour, albumin 2.25g/dL, CRP 1.75mg/L,
proteinuria +3, ANA test +, anti-dsDNA +, and low C3 C4. The assessment was SLE flare, 31/32 weeks of gestation,
single live fetus, unlaboured. She was given methylprednisolone 500mg for 3 days then tapered down, azathioprine
2x50mg/day, chloroquine 1x200mg/day, blood transfusion and albumin infusion. On the 9 th day, she complained of
severe dyspnea and cough due to pulmonary edema. Emergency caesarian section suggested due to acute lung
edema. After delivery (1550g, Apgar score 5-6), she got worse and was on mechanical ventilator. On 12 th day,
antibiotic was added due to pneumonia suspicion. On 17 th day, she had no dyspnea, ESR 75mm/hour, proteinuria
+3. We gave cyclophosphamide 500mg i.v, continued antibiotic and steroid, azathioprine was stopped. On 20 th day,
she was discharged with a good condition.
P2-051 ELEVATED CFDNA IS ASSOCIATED WITH ACTIVE LUPUS NEPHRITIS AND MAY MAINLY DERIVE
FROM NETOSIS IN SYSTEMIC LUPUS ERYTHEMATOSUS
S. Zhang1, G. Wang1, X. Lu1
1Department
of rheumatology, Chian-Japan Friendship Hospital, Beijing, China
Objective. Abnormal formation and insufficient clearance of neutrophil extracellular traps (NETs) has been involved
in lupus nephritis (LN) and can cause the increase of residual NETs in vivo, which is one potential source of
circulating cell-free DNA (cfDNA). This study explored whether cfDNA could be a biomarker for LN.
Methods. Fifty four patients with SLE and 43 age- and sex-matched healthy controls were included in the study.
cfDNA concentration was measured with Picogreen Kit, low-density granulocytes (LDGs) percentage in peripheral
blood mononuclear cells (PBMCs) was tested by flow cytometer and DNase I activity was measured by radial
enzyme-diffusion method.
Results. cfDNA in SLE group was 239.76±57.15 ng/ml, significantly higher than that in healthy control group
(187.96±40.55 ng/ml, P<0.0001). cfDNA in patients with LN was significantly higher than that in patients without LN
(251.8±62.45 ng/ml vs. 213.6±31.34 ng/ml, P=0.028). cfDNA in patients with active LN was significantly higher than
that in patients with inactive LN (260.7±69.8 ng/ml vs. 215.9±29.1 ng/ml, P=0.0349). cfDNA positively correlated with
quantitative 24-hour urinary protein (r=0.350, P=0.013), LDGs (r=0.651, P=0.002) and neutrophils (r=0.584,
P<0.0001), and reversely correlated with albumin (r=-0.500, P<0.0001) and endogenous creatinine clearance rate
(Ccr) (r=-0.354, P=0.044). Compared to control group, SLE group exhibited a significantly increased percentage of
LDGs in PBMCs and a significantly decreased DNase I activity.
Conclusion. Elevated cfDNA is associated with active LN and may mainly derive from NETosis by neutrophils as
well as LDGs in SLE. cfDNA could potentially aid in the prediction of LN risk in SLE patients.
Poster Session: Session 2: Systemic Sclerosis and myositis
P2-052 THE EFFECTIVENESS MESOTHERAPY OF FACE PATIENTS WITH SYSTEMIC SCLEROSIS.
M.L. Sukhareva1, T. Dubinina1, S.H.F. Erdes1, O.V. Pushkova1
1RAMS,
State Institute of Rheumatology V.A. Nasonova, Moscow, Russia
OBJECTIVE: To evaluate the efficacy of mesotherapy (MT) in patients with SSc.
MATERIALS AND METHODS: The study included 22 women 11 pts (MT group) and 11 pts (control group) with SSc,
the average age - 42,2 ± 12,1 years, mean disease duration - 7,5 ± 3,4 years. Medicinal homeopathic preparations
(making Germany) with antioxidant and antifibrotic effect used for injection MT with, the course duration was 10 days.
The efficacy evaluated by Modified Rodnan skin score (MRSS), mouth opening in mm, elastography (EG). The
efficacy MT evaluated at baseline (T0), after 10 days (T1), 3 (T2) and 6 mth. (T3).
RESULTS:
The difference between T0 and T1 in MT group for opening the mouth was 44,3 ± 9,8 mm (p <0,05). MRSS in MT
group was significantly improved in T1 in contrast of control group, and maintained at a level of T2 and T3 (p <0,05
compared to T0). According EG in healthy individuals an area corresponding to the dermis showed a uniform redyellow color, and in patients with SSc - blue, with sporadic inclusions islands of yellow and green. According to the
results EG skin of patients in the MT group T1 was as skin of healthy individuals. The effect is maintained for 3-6
months.
CONCLUSION:
Application antioxidant and antifibrotic by MT in SSc should reduce the density of the skin, increase the oral cavity,
helps to smooth wrinkles around the mouth with saving effect for 3 to 6 months. There was no adverse reactions.
P2-053 PULMONARY HYPERTENSION IN SCLERODERMA
A. Kurniawan1, N. Lugito1, M. Tjiang1, I. Wijaya1, T. Yanto1, D.L. Gaol2
1Internal
medicine, University of Pelita Harapan, Jakarta, Indonesia
medicine, University of Indonesia Christian, Jakarta, Indonesia
Background: Transthoracic Echocardiography (TTE) has emerged as important noninvasive method of studying
cardiac abnormality. There have many report describing the pulmonary hypertension seen in patient with
scleroderma. The study was carried out to detect the cardiac abnormalities in patient with scleroderma.
2Internal
Aim : To know an incidence pulmonary hypertension in scleroderma patients in our country.
Methods: This a descriptive study, we took data retrospectively from medical record, we studied a resting TTE
findings in 19 patients with scleroderma and were evaluated for incidence of pulmonary hypertension.
Results: Following results of TTE in 19 scleroderma patients, 17 (89.5%) patients most were females. Mean age of
the patients was 37.3 ± 12.9 years old. TTE revealed abnormality in 14 patients (73.68%). Of these findings, 2
patients (10.52%) had pericardial effusion. Eight patients (42.10%) had some valvular involvement and there were
two patients (10.52%) had low left ventricular ejection fraction. Four of the 19 patients (21.05%) were diagnosed
having pulmonary hypertension. Some degree diastolic dysfunction was found 8 patients (42.10%). We did not find
any thrombus from TTE in our patient.
Conclusion: The incidence of pulmonary hypertension in our study was 21.05%.
KEY WORDS: Echocardiography, Pulmonary hypertension, Scleroderma.
P2-054 5-AMINOLEVULINIC ACID PREVENTS SKIN AND INTERNAL ORGANS INFLAMMATION AND FIBROSIS
IN MURINE SCLERODERMATOUS GRAFTS-VERSUS-HOST DISEASE, A MODEL FOR HUMAN
SCLERODERMA
C. Liu1, X. Yang2, M. Fujino1, H. Ito3, K. Takahashi3, F. Abe3, M. Nakajima3, T. Tanaka3, Y. Isaka4, H. Rakugi4, H.
Zou2, X. Li1
1Division
of Radiation Safety & Immune Tolerance, National Research Institute for Child Health and Development,
Tokyo, Japan
2Rheumatology, Huashan Hospital Fudan University, Shanghai, China
3Division of Pharma R & D, SBI Pharmaceuticals Co. Ltd., Tokyo, Japan
4Geriatric Medicine & Nephrology, Osaka University Graduate School of Medicine, Osaka, Japan
Background Scleroderma or systemic sclerosis (SSc) is a clinically heterogeneous rheumatologic autoimmune
disease characterized by skin, internal organs and blood vessels, and there is no effective therapy. The purposes of
current study are to develop a model of GvHD-induced scleroderma that more fully represents human condition, and
to investigate the effects of 5-aminolevulinic acid (5-ALA), an intermediate of heme synthesis, enhance HO-1 activity
to cleave heme to form biliverdin, CO, and iron on this model.
Methods SSc-GvHD was induced by injection of lymphocytes from B10.D2 mice into BALB/c mice deficient in mature
T and B cells (recombination activating gene 2-null mice).
Results We successfully established an SSc-GvHD model, which is similar to the human disease particularly in the
skin, progressive inflammation and fibrosis of internal organs including lung, kidney, and liver. We found that
treatment with 5-ALA and iron (Fe2+) significantly reduced progressive inflammation and fibrosis in the skin and
internal organs. Furthermore, by quantitative real-time PCR analysis, 5-ALA and Fe2+ suppressed the inflammatory
cytokines and TGF-beta, type I collagen mRNAs expression. These results indicate that combination treatment with
5-ALA and Fe2+ exhibited a protective effect on tissue fibrosis and inflammation of SSc-GvHD model mice.
Conclusion The model of GvHD-induced SSc has shown most of symptoms of human disease and is likely to
contribute to better understanding of the disease mechanism. Furthermore, efficacy of the 5-ALA has important
implication for clarifying the mechanism of CO and/or HO-1 activity in autoimmune diseases, and may provide a
favorable opportunity for clinical therapy.
P2-055 ASSOCIATION BETWEEN OVERLAP AUTOANTIBODIES AND OVERLAP CLINICAL FEATURES IN
PATIENTS WITH AUTOIMMUNE MYOSITIS IN BULGARIAN POPULATION
V. Reshkova1, D. Kalinova1, R. Rashkov1
1Clinic
of rheumatology, University hospital 'St. Ivan Rilski", Sofia, Bulgaria
Background: Autoimmune myositis (AIM) is a syndrome characterized by involvement of the cellular and humoral
immune systems in skeletal muscle pathology, response to immunotherapies and the presence of autoantibodies Abs
(myositis-specific and myositis-associated autoantibodies) in the serum of some patients. Myositis Abs are
collectively referred to herein as overlap Abs. Although it has been suggested that myositis Abs may identified distinct
disease entities, in practice these Abs often segregate with overlap manifestations 1.
Objectives: The objective of this study was to determine prevalence of overlap Abs and to analyze association
between overlap clinical features and overlap Abs in patients with AIM in Bulgarian population.
Methods: Sera were collected from 80 patients with AIM as diagnoses were determined using Bohan and Peter
classification and Troyanov clinicoserologic classification. Sera were analyzed by Immunoblot test and enzyme linked
immunosorbent assay. Association between clinical manifestation and Abs were analyzed by Chi-square analysis,
FisherÕs Exact test. Positive and negative predictive values, and likelihood ratio were calculated using SPSS 13.0 for
Windows.
Results: Overlap Abs were presented in 70% of patients with AIM. It was found that the presence of overlap Abs was
strongly associated with overlap clinical feaures at AIM diagnosis (specifity 75%, positive predictive value 89.5%,
positive likelihood ratio 3.37). The sensitivity of Abs for overlap features was 84%, whereas the negative predictive
value was 65%.
Conclusion: In the study overlap Abs often associated with overlap manifestation. Therefore it has been suggested
that much of AIM is composed of ovelap myositis.
P2-056 PLASMA DNASE I WITH IMPAIRED ACTIVITY FAILS TO DEGRADE NETS AND IS ASSOCIATED WITH
INTERSTITIAL LUNG DISEASE IN PATIENTS WITH DERMATOMYOSITIS
S. Zhang1, G. Wang1, X. Lu1
1Department
of rheumatology, Chian-Japan Friendship Hospital, Beijing, China
Objective: Interstitial lung disease (ILD) is a poor prognostic factor of Dermatomyositis (DM) and its pathogenesis is
not well known. DNase I unable to degrade neutrophil extracellular traps (NETs) is considered to play an important
role in autoimmune disease. This study focuses on the role of DNase I in the pathogenesis of ILD in DM patients.
Methods: Plasma samples from 36 patients with DM (19 with ILD) and 47 age- and sex-matched healthy controls
were tested for DNase I activity, DNase I inhibitors, formation and degradation of NETs, LL-37 and circulating cellfree DNA (cfDNA). Results from DM patients with ILD (DML) were compared with those from DM patients without ILD
(DMNL).
Results: DM plasma induced more NETs formation than control plasma (246±93.48 RFUs vs. 191.6±52.88 RFUs,
P=0.002). Plasma LL-37 and cfDNA in DM group were 48.7±11.25 ng/ml and 264.9±62.95 ng/ml, respectively,
significantly higher than those in control group (33.81±13.08 ng/ml, P=0.0031; 197.1±31.36 ng/ml, P<0.0001,
respectively). DNase I activity in DM group was 0.3586±0.2166 U/ml, significantly lower than that in control group
(0.5442±0.2464 U/ml, P=0.0003). Furthermore, DML exhibited significantly lower DNase I activity than did DMNL
(0.2607±0.1709 U/ml vs. 0.4638±0.2099 U/ml, P=0.003). Specific DNase I inhibitors were found in 36.84% of DML
and 5.88% of DMNL (P=0.0257). Compared with DMNL, DML cannot degrade NETs completely (83.41%±12.64% vs.
58.58%±21.4%, P<0.0001).
Conclusion: Data show that DML with impaired DNase I activity failed to completely degrade NETs, which suggests
that DNase I may play an important role in pathogenesis of ILD in DM patients.
Poster Session: Session 2: Tolerance: oral, peripheral, Treg, Tregitope
P2-057 MODULATION OF NEURONAL AUTOIMMUNE IMMUNE DISORDERS WITH IRT5 PROBIOTICS
C.C.S. Chae1, H.S.K. Kwon2, S.W.K. Song1, C.J.S. Chun1, I.S.H. Im1
1Life
Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, Korea
and Immunobiology, Harvard Medical School, Boston, USA
Alteration of gut microbiota composition is associated with diverse immune disorders and restoration of dysbiosis in
disease state with beneficial microorganism could confer the health benefits. Recently we have developed a
screening system to selectively identify probiotic strains that could enhance the generation of CD4 +Foxp3+ regulatory
T cells (Tregs). Using the system, we identified a mixture of probiotics (IRT5) that up-regulates Tregs in vivo.
Administration of the IRT5 induced both T-cell and B-cell hypo-responsiveness and down-regulated T helper (Th) 1,
Th2, and Th17 cytokines without apoptosis induction. It also induced generation of CD4(+)Foxp3(+) Tregs from the
CD4(+)CD25(-) population and increased the suppressor activity of naturally occurring CD4(+)CD25(+) Tregs.
Conversion of T cells into Foxp3(+) Tregs is directly mediated by regulatory dendritic cells (rDCs) that express high
levels of IL-10, TGF-beta, COX-2, and indoleamine 2,3-dioxygenase. Administration of probiotics had therapeutic or
prophylactic effects in experimental disease models of inflammatory bowel disease and in non-mucosal immune
disorders such as atopic dermatitis, hapten-induced contact hypersensitivity, rheumatoid arthritis, myasthenia gravis
and multiple sclerosis. The immunoregulatory effect of the IRT5 probiotics is associated with enrichment of
CD4(+)Foxp3(+) Tregs in the inflamed regions. Collectively, the administration of probiotics that enhance the
generation of rDCs and Tregs represents an applicable treatment of inflammatory immune disorders. This study was
supported from the grants funded by the Korea Food Research Institute, GIST-Caltech Collaborative Research
Projects and GIST Systems Biology Infrastructure Establishment Grant funded by the Gwangju Institute of Science
and Technology (GIST).
2Microbiology
P2-058 INDUCTION OF TOLERANCE TO PDC-E2 BY AAV MEDIATED EXPRESSION OF PDC-E2 IN MICE
P. Leung1, T.W. Hsu2, I.H. Lin2, M.E. Gershwin1, S.A. Shu1, P. Wu2, M.H. Tao2
1Internal
Medicine: Rheumatology/Allergy, University of California Davis, Davis, USA
of Biomedical Sciences, Academia Sinica, Taipei, Taiwan
Background and Aims: We have focused on developing unique gene therapy approach to induce tolerance in
primary biliary cirrhosis using a hepatotrophic adeno associated virus vector (AAV8) engineered with a liver-specific
enhancer/promoter to over express PDC-E2 in liver.
Methods : We have designed two AAV-PDCE2 constructs to express the mouse PDC-E2 in either a) the
mitochondrial form (AAV8-E2-flag) or b) the cell membrane form (AAV8-HA-E2mb) and examined their ability to
induce immune tolerance to PDC-E2 in a PDC-E2 immunized mouse model. The mitochondrial and membrane
bound PDC-E2 were tagged with flag and HA epitopes, respectively, to distinguish transgenic PDC-E2 from
endogeneous PDC-E2. AAV8 expressing the firefly luciferase (AAV8-luc) was included as a negative control.
C57BL/6 mice were administered I.V. with either with 1011 vector genome of AAV8-E2-flag, AAV8-HA-E2mb, or
AAV8-luc and thereafter immunized with recombinant PDC-E2 every 2 weeks. Liver specific expression of transgene
were determined by measuring luciferase reporter gene expression using in vivo imaging and ex vivo imaging. Sera
were collected for anti-PDC-E2 determination.
2Institute
Results : Luciferase was highly expressed in livers administered with AAV8/luc whereas only background level of
light emission was detected with AAV8-E2-flag and AAV8-HA-E2mb. More importantly, anti-PDC-E2 was reduced
mice administered with AAV8-E2-flag or AAV8-HA-E2mb when compared with control PDC-E2 only treated mice
throughout. The degree of inhibition was more significant in sera from AAV8-HA-E2mb than controls. Conclusion:
The liver specific expression of the PDC-E2 transgene and successful inhibition of anti-PDC-E2 in both AAV8-E2flag, AAV8-HA-E2mb is unique and a novel strategy for restoration of tolerance
P2-059 DIVERGENCE OF T LYMPHOCYTE SUBPOPULATIONS IN QUAIL-WHITE LEGHORN, DUCK-WHITE
LEGHORN CHIMERAS
Z.D. Li1, L. Yan1, C.Y. Dai1, X.X. Yu1, L. Rui1, X.Y. Zhou1, H.B. Cao1, W.X. Zhang1, Z.Y. Wan1, Q. Shao1, Y.N. Lu1
1Biochemistry
and Molecular Biology, China Agricultural University, Beijing, China
Chimerism, with coexistence of cells from the same or different species, is a reliable model for inducing immune
tolerance. In avian chimeras, development of chimeric organs induces immune tolerance in chick recipient. The
disorder of immune system may lead chimeras to death. In this study, we detected the divergence of T lymphocyte
subpopulations in inter-species avian chimeras to investigate the mechanism of immune tolerance and rejection.
Quail-White Leghorn (WL) and Peking Duck (Duck)-WL chimeras were produced by injecting dissociated
blastodermal cells of quail and duck into WL embryos respectively. T lymphocyte subpopulations in spleen and
thymus from died chimeras on E (Embryonic) 13-E15d, died chimeras on E21d and survival chimeras of two months
were analyzed by flow cytometry. The results indicated that there were no significant differences on CD4 + T
lymphocytes in thymus, but CD4+ T lymphocytes in spleen were decreased in two-months Quail-WL and Duck-WL
chimeras. In Quail-WL and Duck-WL chimeras died on E13-15d, CD4+ T lymphocytes in spleen were significantly
increased. Although there were no significant differences of CD4 + T lymphocytes in thymus, the CD3+ lymphocytes in
thymus were sharply decreased in E21d dead Quail-WL and Duck-WL chimeras. The divergence of T lymphocyte
subpopulations in chimeras may be due to chicken regulatory T cells (Tregs) which can suppress proliferation of
naive CD4+ T lymphocytes to keep chimeras alive. Anti-chicken CD25 monoclonal antibody was prepared and CD25
gene expression was detected in thymus and spleen. There need further evidence to proof the potential regulation of
Tregs in avian chimeras.
P2-060 ADIPONECTIN REDUCES HEPATIC ISCHEMIA REPERFUSION INJURY BY SUPPRESSING
INFAMMATORY REACTIONS IN RATS
J. Li1, S. Nie1
1the
Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
Background and aims: Liver transplantation has been proven to be a successful treatment to end-stage liver
disease. However, immune rejection, ischemia-reperfusion (IR) injury is an important problem that contributes to graft
nonfunction or early dysfunction, which significantly affects both morbidity and mortality. In the present study, we
investigated the role of adiponectin, an adipokine mainly secreted from adipocytes, in attenuating ischemiareperfusion injury.
Methods: Sprague Dawley rats were subjected to 60 min of partial hepatic ischemia followed by reperfusion. Before
the induction of IR, rats were treated by an intravenous administration of either adiponectin (Adiponectin group) or
phosphate buffered saline (PBS) (Control group), the sham group received no treatment. Hepatic functions, mRNA of
inflammatory cytokines in liver tissue, the serum levels of related cytokines, apoptosis of hepatocyte and histological
examination were assessed.
Results: At 6h after reperfusion, the serum levels of hepatic enzymes in the Adiponectin group were signi?cantly
lower than the Control and Sham group (P<0.05). MRNA of inflammatory cytokines and the serum levels of
inflammatory cytokines expression were also suppressed. Furthermore, Adiponectin also signi?cantly reduced
hepatocyte apoptosis and NF-κB activity at 24h after reperfusion. The histopathology showed mild changes in the
Adiponectin group.
Conclusion: Adiponectin has a protective effect against in?ammatory reactions, and reduces hepatocellular injury
induced by hepatic IR in rats.