1
Early nutrition and immunityprogress and perspectives
Sonja Lang
Katja Bohländer
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
2
Overview
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•
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•
•
•
•
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Tolerance
Role of nutrition
Feeding practises
Role of dendritic cells, lactobacilli
Intestinal colonization
PUFA
LCPUFA
Lipid rafts
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
3
Immunological tolerance
• Lifelong processes
• Polarization of Th cells: Th2*, Treg ↑↑
• *Recognition of ultra-low antigen dosis
(IgE, IgA)
• Sterile GIT
• Exposure to bacteria at term and after
(mother´s skin, breast milk  maturation
of infant´s gut
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
4
Nutrition + immunologic development
Nutrition
• …might affect ID during pregnancy, suckling period,
introduction of formula and solide foods 
• …source of antigens IS must become tolerant
• …provides factors, which modulate immune
maturation + responses + influences intestinal flora 
antigen exposure, immune maturation, immune
response
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
German Infant Nutritional
Intervention Study
• Effects of hydrolysed and standard cow´s milk formula
• human milk feeding:
• Hydrolysed formula:
↓ allergic diseases at 1y
↓ atopic dermatitis
-
• Extensively hydrolysed formula: allergy preventive effect
• Partially hydrolysed formula: + allergy preventive effect
• Keeping pets (dogs!)  atopic diseases ↓
• Caesarean section: different gut flora, antibiotics  diarrhoea,
allergic sensitization↑
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
5
6
Dendritic cells + Lactobacilli
Function of DC:
drive differentiation of naive Th cells into
Th1, Th2 or Treg cells
- Treg cells: prevention of autoimmunity,
allergy
- L. reuteri + L. casei prime human DC and
drive development of Treg cells by
targeting DC-SIGN
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
IMMUNOFLORA study
7
• „…how early intestinal colonization affects the
development of putative Treg cells and clinical allergy
in Swedish infants“
• Western infants have a delayed acquisition of several
gut microbes and a reduced turnover of strains in
intestinal flora Exposure ↓, variety ↓ of
environmental bacteria
• Early food allergy ↔ poor colonization with S. aureus
(strong T cell stimulation)
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
PUFA
•  in the intake of saturated fatty acids
•  in the intake of n-6 family of PUFA
• Linoleic acid
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
8
9
N-6 family of PUFA
Linoleic Acid
Arachidonic acid
Prostaglandine
Leukotriene
Tromboxanes
PGE2
LTB4
TXA2
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
4-series leukotrien
= mediators of allergic inflammation:
• Vascular permeability
• Leucocyte chemotaxis
• Respiratory burst
• Production of inflammatory cytokines
• HYPOTHESIS:  intake of linoleic
acid  prevalence of
atopic disease
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
10
n-3 family of PUFA
• -Linolenic acid
EPA
DHA
 Increased consumption:
→ incorporation into immune cells
→ decrease the production of
prostaglandin E2 and
other eicosanoids
 Protective towards allergic disease
 E.g. n-3 LCPUFA status was lower in cord blood serum from
pregnancy of allergic compared with non allergic mothers.
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
11
n-3 family of PUFA
12
• Positive results in patients
with asthma
• n-3 LCPUFA intervention:
Stronger impact on fetal and
neonatal Th1/Th2 immune responses compared to
immune responses beyond early infancy
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
n-3 LCPUFA
• Influence on T-cell functional responses and
signalling
• First, prostaglandin E2 influence the activity of DC,
differentiation of naive
T-cells and activity of Th1 and Th2 cells
• Second mechanism:
Direct alteration of gene expression through
modification of transcription factor activity
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
13
n-3 LCPUFA
• EPA and DHA give rise to a novel family
of eicosanoid-like mediators, called Dand E- resolvins
• Inhibition (in vitro):
– T-cell proliferation,
– Production of IL-2 and IFN-
– Surface expression of CD25
18.12.2006
Immunologisch relevante Aspekte von
Lebensmittel, Probiotika und
Nutrigenomics, Dr. Alexander Haslberger,
Evaluation of Allergenicity of Genetically
Modified Foods
Report of a Joint FAO/WHO Expert Consulation on
Allergenicity of Foods Derived from Biotechnology
22 - 25 January 2001
Rome, Italy
Introduction
• 29 May to 2 June 2000 Joint FAO/WHO
Geneva, Switzerland
• follow-up: 22 to 25 January 2001
Rome, Italy
members: 28 experts and authors of
discussion papers
Allergenicity
• Most frequently asked questions
– safety of genetically foods
• reliable methodology to assess the
allergenicity of foods produced by the
recombinant DNA technique needed
Scope
• General consideration of allergenicity of
genetically modified foods
• Consideration of the decision-tree approach
• Specific questions arising in relation to the
assessment of allergenicity of genetically
modified foods
Food Allergies
„Overhwhelming pathological reactions of the
body due to intercurrent contact with
antigens“ Clemens von Pirquet 1906
• IgE-mediated allergy
• Cell-mediated allergy
• Oral allergy syndrome
Decision tree
• Criteria
– source of the transferred genetic material,
– molecular weight,
– sequence homology,
– heat and processing stability,
– effect of ph and/or gastric juices and
– prevalence in foods.
Source of gene allergenic
Yes
No
Sequence
Homology
Sequence
Homology
Yes
No
Specific
Serum
Screen
Yes
Target Serum
Screen
No
Yes
Yes
No
No
Pepsin
Yes
Resistance
Yes
&
Animal Models
Likely
Allergenic
+/+ +/- -/High
Low
Probability of
Allergenicity
Post marketing surveillance
• Traceability and labelling
• Lack of background data
• Many confounding food and non-food related
factors
• Changes in diets over time
• Lack of trained experts an infrastructure
Other criteria
• Level of expressions
• Unintended effects
Evaluation of Allergenicity of Genetically
Modified Foods
Martina Pomper
9603177
1. Risk assessment and food allergy: the
probabilistic model applied to allergens
Spanjersberg, M.Q.I., Kruizinga, A.G., Rennen, M.A.J., Houben, G.F.,
Food Chem Toxicol, 45: 49-54 (2007)
Ines Pree, Immunology and Food, WS 2006
The probabilistic approach in food allergy
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Purpose: avoidance of hidden or undeclared allergens
 Risk assessment
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Conservative determinstic appraches: worst case value „an allergic
reaction cannot be excluded“
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Probabilistic approach quantifies health risk asessment by
1.
Hazard identification: situation, symptoms, target organs
2.
Hazard characterization: threshold, minimum dose
3.
Exposure assessment: intake etc.
4.
 Risk assessment
Ines Pree, Immunology and Food, WS 2006
Hazelnut allergens in chocolate - a case study
Risk assessment of three bars, each of a different brand, according to:
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Prevalence
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Threshold (LOED*)
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Consumption pattern
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Allergen concentrations
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Computer software
*lowest observed eliciting dose
Result:
The allergen was detectable in each bar but at different concentrations.
Ines Pree, Immunology and Food, WS 2006
The probabilistic vs. the deterministic approach
• The deterministic model does not distiguish between the
different degree of contamination.
“An allergic reaction cannot be excluded” is true for all three brands.
• The probabilistic model gives more detailed information and
avoids overestimation of the risk for the population.
1.
All three brands together: highest mean risk of 0.05%, i.e. less than 500 subjects per million
will respond.
2.
The risk for breakfast consumption is higher when compared to lunch. There was a lower
risk for women, since men consume more.
3.
Brand 3: the highest risk of 0.004%; less than 40 subjects will respond which reflects a
lower contamination of brand 3
Ines Pree, Immunology and Food, WS 2006
2. Practical and predictive
bioinformatics methods for the
identification of potentially crossreactive protein matches.
Goodman, R.E. Mol Nutr Food Res, 50: 655-660 (2006).
Ines Pree, Immunology and Food, WS 2006
Potential allergenicity in GE food
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If the protein similar to a known allergen, specific IgE may
be cross-reactive (recognition of similar epitopes)
sequence  conformation  cross-reactivity
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How to determine potential allergenicity:
1.
2.
3.
Compare amino acid sequences by computer programs
Recruit potentially at-risk individuals (allergic patients)
Perform serum testing, skin prick testing, food challenge.
Ines Pree, Immunology and Food, WS 2006
Comparison of amino acid sequences
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FASTA and BLAST alignments (used for species homologies) to identify
IgE and T cell epitopes?
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Since 1990ies: 8 contiguous amino acid matches
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In 2001: 6 amino acid matches are too short, too many matches; >35%
identity over 80 amino acids is useful
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Points of discussion:
1.
Allergen databases are incomplete, mainly lacking minor allergens
2.
Epitopes are poorly defined and the relevance of conformational epitopes is not
fully established
3.
Analysis of 3D structures: group proteins into structural families and compare
motif recognition patterns
Ines Pree, Immunology and Food, WS 2006
Consensus 2005- workshop in Spain
• Short matches are not predictive
• FASTA and BLAST algorithms are efficient
• Structural comparison may be very useful
• There are currently no data to change the guidelines (>35%
identity over 80 amino acids)
Ines Pree, Immunology and Food, WS 2006
Summary/ Conclusion
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In risk assessment of food allergens the current precautionary “may
contain” labelling is based on the possible presence of an allergen
rather than on the assessment of a quantative risk. The quantative
expression of risk could avoid unnecessary labelling or recalls.
•
In the prediction of IgE cross-reactivities in food allergy: structural
comparison may be useful. However, there is currently not enough
data to change current guidelines (>35% identity over 80 amino
acids).
Ines Pree, Immunology and Food, WS 2006
Immunity, Inflammation and
Allergy In The Gut
Thomas T. MacDonald and Giovanni
Monteleone
The gut
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•
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(1)
Nutrients get absorbed
Potential to compromise host defense
infection diseases are largely under control
But: gastrointstinal food allergies have increased
 Probably because of the absence of gut infections
has upset the balance between the commensal in
the gut
The gut
(2)
• High active immunsystem
• Barrier is a single layer of epithelium
• No completely prevent of antigens entering the
tissue
• Several mechanism how antigens get trough the
epithelium
 Immunsystem gets constantly activated
Components of the Immunsystem in the gut
• Pattern recognition receptors – recognize conserved
structures
• Severals receptors like TLR, NOD,..
• Recognition of TLR ligands increases gut barrier
function
• Hsp25 and hsp70
• CD4+ T cells
• Macrophages
• Dendritic cells
Activation
• B and T Cells activated
 Expression of α4β7 integrin
• TLR or NOD activate NFĸB
 Leads to pro-inflammatory gene expression
• Chemokine fine-tune the localisation of the tissue
Crohn‘s disease (1)
• Complex genetic disease
• Mucosal ulceration, ulcers penetrate into the gut wall
• Antigen is not yet identified
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Isolated CD 4+ TH1 cells produce large amount of interferon γ
Overexpression of transcriptionfactor T-bet
Macrophages produce large amount of TH1 inducit cytokines
T-cells show resistence to apoptotic signals and have an
increased cell cycle
Crohn‘s disease (2)
• Genes located on the chromosomes 1, 5, 6, 12, 14, 16
and 19
• Different polymorphism in the Nod2 gene
 Mutations in the Nod 2 can lead to a decreased ability to
kill gut bacteria
• OCTN and DGL5 gen
 Important for epithelial permeability
 Disruption leads to inappropriate exposure of the
mucosal immunsystem to bacterial products
Celiac disease (1)
• In some genetically susceptible individuals
after ingestion of cereal products
• Treated by adherence to a gluten free diet
• morphological chances to the mucosa of the
upper bowel – long crypts and atrophy of villi
Celiac disease (2)
• 4 components involved
à Gluten is prolin and glutamine rich, has negatively
charged residues
à tTG deaminates glutamin to glutamic acid and produces
negatively charged residues
à Necessary for efficient binding to HLA-DQ2 and
furthermore activation of gluten specific t-cells
à Peptides of gliadin activate gut macrophages to produce
IL-15 -> increases MICA and arms IEL to kill MICA and
epithelial cells
Control of inflammation in the gut
• T-cells involved in tolerance against
commensals
• Commensals which crossed the barrier will be
phagocytosed without cytokinproduction
 The T-cells die by apoptosis
§ The epithelial permeability is genetically
determined
 Importend factor in the developement of
diseases
Probiotics
Do They Help to Control
Intestinal Inflammation?
Probiotics
• In the maintenance therapy for the inflammating
bowel diseases, Crohn’s diseases and ulcerative
colitis
• Specific molecules modulating defined targets in
the gut mucosal and systemic immune system
(Active) Ulcerative Colitis
• Ulcerative Colitis: Study comparing mesalamine
treatment with E.coli Nissle 1917 treatment
• relapse rate were not different between groups
à E.coli Nissle 1917 is a safe alternative for prevention
of relapse in ulcerative colitis
§ Active Ulcerative Colitis: Trial examining the
effectiveness of the fermented milk containing
Bifidobacteria strains and Lactobacillus acidophilus.
 Remission was achieved in 4 of 12 patients
Pouchitis
• Study of the ability of VSL 3 to prevent
recurrence of chronical relapsing pouchitis
17 of 20 patients remained in remission
§ But: Probiotics have failed to demonstrate
efficacy
Crohn‘s disease
• Pioneer study to examine the efficacy of E.coli
Nissle 1917 in maintaining remission in
Crohn’s diseases
 Groups did not differ in the rate of
remission regardless of disease location
Conclusions
• More effective in preventing relapse of inflammation
than suppressing diseases
• In active inflammation sufficient data are missing
§ Genetically engineered bacteria delivering antiinflammatory cytokines or other biological
molecules
Allergy, Parasites and the
Hygenie Hypothese
Maria Yazdanbakhsh
Peter G. Kremsner
Ronald van Ree
Atopy and Allergic diseases
• significant increase in pervalence of allergic deseases in the
last 20-30 years
• differences in developing and industrial countries
• risk faktors such als increased exposure to indoor allergens
• childhood infektions shows a negative association with atopy
and allergic diseases
Atopy
• enviromental allergen leads to T cell stimulation
• release of Cytokines (IL4, IL5, IL13)
• raised IgE levels
à Increased numbers of eosinophiles and mast cells
Hygenie Hypothese
•
•
•
•
High hygenie
Vaccination
Antibiotics
Limited exposure to pathogens during early
childhood
 Can lead to atopies and allergies
Helminth infections
• Stimulates TH2 Immunresponses
 High levels of IgE, eosinophiles and mast cells
• People with helminth infections are rarely
afflicted by allergic diseases
 TH2 can‘t be the sole factor for allergic attack
IgE blocking hypothesis
• Highly not specific polyclonal IgE
• Unspecific IgEs satures the Fc-receptors on
mast cells
à The binding site is blocked
à degranulation is inhibited
à Hypersensitiviy will be immediated
Blocking antibodies
• Parasites specific IgG4 antibodies inhibit IgE
mediated degranulation of effector cells
à Possible mechanism of allergen
immunotherapy
à IL-10 stimulates the IgG4 differentation
à Allergic individulas express lower levels of IL10
True or False? The Hygenie
Hypothesis for Crohn‘s disease
Bret A. Lashner M.D.
Edward V. Loftus Jr.M.D.
• Lack of exposures to enteric pathogens makes
one susceptible to Crohn‘s disease
à Multiple childhood infections and poor
hygenie protects
 Host develops tolerance or immunity to
agents that could trigger Crohn‘s disease
Crohn‘s disease
• 2 different studies came to very different conclusions regarding the
hygenie hypthesis
• The results of one study supports the hygenie hypothesis
à Exposure to pathogens in childhood stimulates the immune system
• But the other contradicts
à Poor hygenie may contribute to the pathogenesis
à much work still needs to be done to determine if childhood exposure
are truly important to the pathogenesis of Crohn‘s disease
The PRODIA study
Ann N Y Acad Sci 1079: 360-364 (2006)
Probiotics for the Prevention of Beta cell Autoimmunity
in Children at Genetic Risk of Type 1 Diabetes
Section of a pancreas of a dog
Source: Gray´s anatomy
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Diabetes mellitus
Aretaeus of Capadocia: diabaínein „passing through“ or „siphon“
Thomas Willis (1675): mellitus „sweet taste“
Chronic disorder of carbohydrate, fat and protein metabolism caused by
lack or non-functionality of insulin
• Hyperglycemia  Polyuria, excessive thirst, polyphagia, weight loss,
fatigue, blurred vision, muscle cramps, nausea
• Ketoacidosis
• Nonketotic hypersomolar coma
• Retinal damage
• Chronic renal failure
• Diabetic neuropathy
• Coronary artery disease
• Gangrene: „Diabetic foot“
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Type 1 Diabetes mellitus (T1DM)
A.k.a Insulin-dependent diabetes mellitus (IDDM) or juvenile diabetes
Loss of the insulin-producing β-cells of the pancreas leading to deficiency of insulin
Type 2 Diabetes mellitus (T2DM)
A.k.a. Non insulin-dependent diabetes mellitus (NIDDM) or adult-onset diabetes
Combination fo defective insulin secretion and defective responsiveness to insulin
Type 3 Diabetes mellitus (T3DM)
A.k.a. Gestational Diabetes
Reduced receptivity to insulin of pregnant women due to their hormone status
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
The Islets of Langerhans
Paul Langerhans 1869
One million islets/pancreas
Combined weight 1-1,5 gramms (1-2% of pancreatic mass)
~1000 cells/islet
• 65–80% β-cells producing Insulin and Amylin
• 15–20% α-cells producing Glucagon
• 3–10 % δ-cells producing Somatostatin
• 1% PP-cells producing pancreatic polypeptide
Porcine Islet of Langerhans
Brightfield Hematoxylin / Immunofluorescence
Etiology of T1DM
Genetic factors
Prevalence in the general population: 0,2–0,4%
Concordance rate in monozygotic twins: 40-50%
Concordance rate in dizygotic twins and siblings: 5-10%
Genetic susceptibility accounts for ~half of the etiology
Eighteen Loci (IDDM1-IDDM18) identified by positional cloning.
All except four have turned out to be statistical artifacts due to
underestimation of the sample size required for meaningful statistical power
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Genetic factors
1. HLA Class II (IDDM1 Locus)
40-50% of the genetic risk
Predisposing haplotypes:
DRB1*0301(DR3)-DQA1*0501-DQB1*0201(DQ2)
DRB1*0401(DR4)-DQA1*0301-DQB1*0302(DQ8)
Heterozygosity DQ2/DQ8
Protective Haplotypes:
Horm Res 2005;64:180-188
DRB1*1501-DQA1*0102DQB1*0602(DQ6)
Molecular mechanism:
Absence of aspartic acid at position 57 of the β chain of the DQ molecule reversing
the electric charge of the peptide binding groove and altering the binding of epitopes
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Genetic factors
2. INS-VNTR (IDDM2 Locus)
Horm Res 2005;64:180-188
Polymorphism in the 5´flanking region of the insulin gene, consisting of a
variable number of tandem repeats. Either 30-60 repeats (class I) or
120-170 repeats (class II). Homozygosity for class I confers a relative risk
of 2-3 compared with the dominant protective presence of class II.
Probably due to lower thymic insulin levels  hampered negative selection
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Genetic factors
3. PTPN22
Protein tyrosine phosphatase, nonreceptor type 22
The nonreceptor tyrosine phospahatase Lyp is specific to lymphocytes and
suppressesT-cell activation by dephosphorylating three kinases important to
T-cell signaling.
The R620W SNP has also been associated
with other autoimmune diseases like
Rheumatoid Arthritis and SLE
R620W maps to a solid 293-kb
linkage disequilibrium block containing
6 other known genes and 625 known SNPs.
Horm Res 2005;64:180-188
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Genetic factors
4. CTLA-4
Cytotoxic T-lymphocyte-associated antigen 4
Horm Res 2005;64:180-188
Encodes T cell receptor that mediates apoptosis in activated T cells
Associations also in Graves disease and autoimmune hypothyroidism
Functional mechanism of the A6230G polymorphism is unknown.
Contribution of the locus is low (relative risk ~ 1,2).
Environmental factors
• Early exposure to cow milk and gluten
• Aberrant development and maturity of the gut immune
system
• Enterovirus and Rotavirus infections
• Vitamin D3 deficiency
• Toxins: Rodenticides (Vacor), chemotherapeutic agents
(Streptazotocin)
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
T1DM and the gut
• In animal models the incidence of T1DM is highest
in a low microbial load environment
• Diet modifies the development of T1DM in animal models
• T cells from human diabetic pancreas show mucosal
homing properties
Microbiotic colonization of the newborn´s gut by bacteria may be important
for the initial regulation of the developing immune system. Development of
T1DM is associated with intestinal immune activation and enhanced
immunity to food antigens.
Probiotics have been shown to support the development and maturity
of the gut immune system and could therefore support oral tolerance and
protection against enteral virus infections.
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Autoantibodies in T1DM
Type 1a patients: Autoimmune, autoAbs present
Type 1b patients: No evidence of autoimmunity
3 major anti-islet autoantibodies:
• Glutamic acid decarboxylase (GADA): Useful marker
for confirming etiology in long-standing cases
• Tyrosine phosphatase (IA-2A)
• Insulin (IAA): The only β-cell specific autoAg, more in DR4
Most children developing T1DM are positive for at least 2 of these markers.
But B cells apparently not strictly required in the autoimmune process.
Immune dysregulation in T1DM
There has no primary diabetogenic autoantigen been found yet!
Trigger ??
Bacterial products?
Regulatory Th2 cell anergy?
Immunity, Vol 7, 727-738
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
The PRODIA study
Faculty of Health Sciences, Linköping University, Sweden
Pilot study to test feasibility and safety of the protocol. Start in February 2003.
The aim of the main study will be to determine whether the use of probiotics
during the first 6 months of life decreases the appearance of β cell
autoantibodies in children with genetic risk for Type 1 Diabetes mellitus.
Affected factors involved could be the reduced occurence of
enteral virus infection, enhanced maturation of the gut immune system,
reduzed immunization to dietary insulin, or induced immune regulation
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Study design
1. Selection procedure
Double-blind randomized placebo controlled study
February 2003 to June 2005: Inform all parents to
newborn infants at Linköping University Hospital
Informed consent by parents for 1200 children (~60%)
Screen for HLA risk genotypes
(presence of risk alleles without protective haplotypes)
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Study design
2. Treatment
Randomize participants to receive probiotics or placebo
Lactobacillus rhamnosus GG (5 x 109 cfu)
Lactobacillus rhamnosus LC705 (5 x 109 cfu)
Bifidobacterium breve Bbi99 (2 x 108 cfu)
Propionibacterium freudenreichii ssp. Shermani (2 x 10 cfu)
Distributed once a day by parents at home in soluble capsules
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Study design
3. Readouts
Blood samples taken at 6,
12, and 24 months of age
Fecal samples taken at
home at 3 months interval
β-cell autoAbs
Microbiological analyses to
Monocyte and T cell-derived cytokines
detect enterovirus infections
Intracellular signal proteins
Analysis of compliance
(T bet, STAT-4, STAT-6, GATA-3)
Monocyte activation markers
upon LPS and THA stimulation
PHA and insulin-dependent T cell responses
Isolation of enterovirus RNA
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Results
264 children with risk genes among the 1200 participants
1/168 IAA-positive at 6 months of age
1/61 GADA-positive at 24 months of age
1/61 IA-2A positive at 24 months of age
Expected: ~2% prevalence of at least one of the autoAbs at 24 months
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Conclusions
„The detected number of autoAbs is close to the expected and there
is no evidence that the intervention would increase the appearance
of beta cell autoimmunity in the children who participate in the study.“
„The PRODIA study protocol seems to be safe and the study protocol
is feasible for the families.“
Mechanistic studies about the development of the immune system and
the occurence of eneterovirus infections are ongoing
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics
Markus Hoffmann
Wirken Phytoöstrogene immunmodulatorisch?
Präsentation im Rahmen der LV
Immunologisch relevante Aspekte von
Lebensmitteln/Nutrigenomics
WS 2006
Nina Zimbelius
Philipp Schatzlmaier
Phytoöstrogene/Isoflavone
•
Soja ist Hauptquelle für Genistein, Daidzein, Equol
•
Strukturell ähnlich zu 17-Östradiol
•
Binden an ER, ER
•
Ziele: Reproduktions-, Immunorgane
•
Bestandteil von natürlicher Diät (v.a. Asien)
•
Ebenfalls in modernen Nahrungsergänzungsmitteln (vielfache Dosis)
•
Bestandteil von Babynahrung (soy-based infant formula)
•
Potentieller Einfluss auf Immunsystem durch zahlreiche Studien untersucht
•
Mäuse, Ratten, Menschen
•
Ergebnisse unvollständig, teils widersprüchlich
80
Humanes Östradiol vs. Soja-Genistein
Estradiol (a human estrogen) and genistein (a phytoestrogen)
The similarly-placed hydroxyl groups at both ends of these two molecules allow
them to bind to human estrogen receptors.
81
Immun-inhibitorische Effekte von Genistein
•
Protein Tyrosine Kinase Inhibitor
•
NO-Produktion in Makrophagen 
•
T-Zellen-Proliferation durch CD28-AK 
•
Interleukin-Produktion , TCL tumorizidale Aktivität 
•
Leukocyten-Adherenz , T-Zellen-Motilität 
•
Aktivierung von NK-Zellen durch LPS 
 Vorsicht: in vitro Ergebnisse bei teils sehr hohen Konzentrationen (100µM)
historische Diät: < 1µM im Serum bei japanischen Erwachsenen
effiziente Aufnahme bei Kleinkindern: ca. 4 µM
keine Abnormitäten im Erwachsenenalter dokumentiert
allerdings sensibles Alter für Thymusentwicklung
82
in vivo Ergebnisse bei Mäusen und Ratten
83
Effekte von Östrogenen auf T-Genexpression
• Ovariektomisierte Mäuse-Babies
• Gabe von E2 und Genistein
• Genexpression durch DNA arrays untersucht
• Gene für Transkription, Apoptose, ZZ beeinflusst
• E2 eher , Genistein eher , grosses overlap an Genen
• Genistein wirkt auch auf E2-nicht-responsive Gene
• Down-Regulierung von CD4-Transkript
84
Ernährungsstudie beim Menschen
•
23 Männer und 18 Frauen, Hypercholesterinämie, 62 J.
•
Post-Menopause (Ersatztherapie!)
•
Drei kontrollierte Diätphasen à 1 Monat:
a) low-fat Kontrollphase
b) high isoflavone soy phase (73mg/d intake)
c) low isoflavone soy phase (10mg/d intake)
•
Am Anfang und Ende jeder Phase wurden bestimmt:
Körpergewicht, Blutdruck, Lipoproteine/Blutfette
Proteine der akuten Phase im Serum: CRP, SAA
proinflammatorische Cytokine im Serum: IL-6, TNF-
85
Ergebnisse Entzündungswerte
86
Schlussfolgerungen
• Geschlechts-spezifischer Effekt
• IL-6 , immun-stimulatorisch
• Negativ: Autoimmunität, CHD
• Positiv: Antwort bei Infektionen, Tumor defense
• IL-6 reduziert Plasmakonzentrationen von ILGF-1
• Geringere Mortalität bei Hormon-abhängigen Tumoren in Asien, auch bei
Brustkrebs
• Östrogene & Isoflavone wirken antioxidativ
• Vermeidung von DNA damage > Anti-cancer effect
87
Referenzen
• Cooke, P.S., Selvaraj, V., and Yellayi, S. (2006) Genistein, Estrogen
Receptors, and the Acquired Immune Response. J. Nutr. 136: 704-708
• Jenkins, D.J.A., Kendall, C.W.C., Connelly, P.W., Jackson, C.C., Parker, T.,
Faulkner, D., and Vidgen, E. (2002) Effects of High- and Low-Isoflavone
(Phytoestrogen) Soy Foods on Inflammatory Biomarkers and
Proinflammatory Cytokines in Middle-Aged Men and Women. Metabolism
51(7): 919-924
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Nutritional Genomics
genes
nutrients
diet
molecular processes
health
Dietary Factors
• direct and indirect influence
• transcriptional, translational and
posttranlational control
Intestinal Lumen – mucosal
immune system
nutritional environment
hormone - dependent
hormone –
independent:
> nutrients
gene regulation
Nutrigenetic and nutrigenomic effects
• nutrigenetic effect:
influence of polymorphisms on altering the
response to dietary components
• nutrigenomic effect:
ability of different food components to
increase or depress gene expression
FABPs – fatty acid binding proteins
• lipid balance
• control of metabolic and inflammatory
pathways
• modulation of FABP activity
> regulation of lipid-sensitive pathways??
Cancer prevention
• ω-3 Poly Unsaturated Fatty Acids (PUFAs)
> influence on expression of genes
> anti-cancer activity
> downregulation of synthesis and
expression
> induction of pro-apoptotic proteins
Leptin
• communicates information of the bodies fat
stores
• altered levels of leptin > eating disorders
(anorexia nervosa)
• regulates different genes (SCD1)
> leptin resistance in obese individuals
Interleukin 1 genetics,
inflammatory mechanisms,
and nutrigenetic opportunities to
modulate diseases of aging
Kenneth S. Kornman
Inflammation
healthy person
genetics
smoking
body mass index
inflammatory mediators
> concentration
no disease
disease
nutrition
complex chronic disease
Interleukin 1

IL-1 and TNF-
>
early activated

drugs that block their activity
> treatment of rheumatoid arthritis
Interleukin 1 gene variations
Interleukin 1 (IL-1) single-nucleotide polymorphisms
IL-1A
IL-1 cluster
haplotype
IL-1B
(+4845) or
IL-1RN
(-511) or
(+2018) or
(-889)
(+3954)
(-31)
VNTR
1
2
2
1
1
2
1
1
2
1
2b
1
1
2
2
3
1
1
1
1
Increased risks
• haplotype 1
>
periodontitis
• haplotype 1
>
cardiovascular disease
• haplotype 2
>
gastric cancer
Nutrients interact with inflammatory
genes
• poly unsaturated fatty acids (PUFAs)
> inhibition of secretion of IL-1 and TNF-
• nutrients that alter the oxidation-reduction
status of the cell
• different effects in individuals with different
polymorphisms
Conclusion
• better understanding of polymorphisms
> identify at-risk persons > interventions
• screening for bioactive nutrients
• problems:
> costs (testing of asymptomatic people)
> primary preventive measures