GVNMunichNotes - Global Virus Network

Global Virus Network – Spring 2013 Meeting in Munich

Meeting Notes*

*We are grateful to our local hosts at TUM for preparing this meeting summary. Individual speakers have not reviewed or edited their respective sections; the notes may therefore not fully reflect individual speaker comments. Any omissions or inaccuracies are unintentional. --- With thanks, GVN

Thursday, May 30, 2013

Workshop on Durability of Vaccine-induced Immune Response

Lecture Hall: “Pavillon”, Technische Universität München, Klinikum rechts der Isar;

Entrance via Lecture Hall Building: Max-Weber-Platz /

Einsteinstrasse; 81675 München

10:00-10:15 Welcome: Ulrike Protzer, Munich & Sharon Hrynkow, GVN

Introduction: Robert Gallo, IHV

Ulrike Protzer and Sharon Hrynkow welcomed all participants and thanked the many sponsors for the meeting.

Bob Gallo: Durability of vaccine-induced immune response is a neglected question in funding.

Antibodies to HIV envelope don´t last more than 3 months, only against core protein. Why is that? This must be solved in a very novel way. This is why the field needs B-cell immunologists. In monkey model we already get sterilizing immunity against up to 4-5 challenges but not more/longer. So it is still important to improve the durability of this response.

10:15-12:15 1 st

Question: Why do we need sustained vaccine responses?

Chair: Billy Hall, CRID-Ireland; Michael Roggendorf, University of Duisburg-Essen

Christian Mandl, Novartis Vaccines and Diagnostics, Vice President

Alphavirus-Vectored Vaccines: From VRP to SAM ® Vaccines

Viral Vectors provide a number of advantages as vaccine candidates:


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

In vivo expression

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Comprehensive immune response (durability similar to live vaccine?)

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Platform for multiple and complex antigens

Virus-like replicon particles (VRP) derived from Alphaviruses are a single-cycle infection system

Production of alphavirus replicon particles: Transfection of RNA into helper cell line

Example:

CMV Phase I Clinical Trial designed by alphaVax, Inc.

CMV vaccine candidate: bivalent virus-like replicon particle vaccine

Results of Phase I study show good antibody titers and also T-cell response measured by IFNg Elispot

Non-viral delivery of self-amplifying mRNA vaccines:



The vision: Reinvent the gene vaccine



Develop a synthetic vaccine with in vivo properties of antiviral vectored vaccine



Without the complications of cell culture production and anti-vector immunity



That is safe, scalable and widely applicable (platform)

Conversion of the viral delivery to a synthetic, safer delivery system. For this get rid of viral structure proteins>Lipid nanoparticle (LNP) “SAM” vaccine

How do SAM vaccines work? Synthetic vaccine without the complications of a packaging cell line, contamination with replication competent virus and anti-vector immunity.

Mechanism of non-viral delivery of SAM vaccine: Endocytosis, innate immune response?

Major issue: How do we produce it? Can we?

RNA-synthesis: Enzymatic transcription reaction from a DNA template

Producing RNA at a commercial scale does not require cell culture, complex purification, or novel equipment.

1. Cell-free synthesis, RNA is produced from a DNA template using an enzymatic transcription reaction

2. Purification

3. Sterile filtration 0,2um filter

4. RNA drug substance

Delivery: Encapsulation with lipid nanoparticles (LNPs) As used in siRNA studies

Delivery system protects RNA from RNases similar to viral envelopes


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In vivo data: LNP delivery enhances gene expression of embryonic alkaline phosphatase (SEAP); RNA encapsulated in LNPs work better than alone or just mixed

RSV is one of the viruses where a vaccine would be desirable. RSV-SAM vaccine induced good IgG titer in mice. Protects cotton rats from RSV challenge.

HIV SAM vaccine expressing HIV-1 gp140 is potent in NHPs 6/6 rhesus macaques seroconverted after the 2 nd vaccination: 3time SAM, VRP prime and protein boost. Gives good antibody response

CMV: SAM can be used to deliver complex antigens. Several promoters expressing several antigens

(gH gL); good titers in mice

Summary: proof of concept achieved for the SAM vaccines



Potential as a platform technology to address multiple disease targets



Immune responses are comparable to a viral vectored vaccine



HIV-SAM vaccine is immunogenic in NHPs at a 50ug RNA dose

Question: Bob Gallo: Challenge in Rhesus monkeys? Not done yet!

If you induce a T-cell response, problem is you also produce more target cells for HIV! So protective effect might be lost again.

Michael Roggendorf: Do particles go to right cells? Important that they go to dendritic cells: Answer: Go to muscle cells, do we really need to target dendritic cells? Nonetheless, targeted Liposomes are a vision.

Can you use Adjuvants when you use liposomes? Answer, not tried yet. But the RNA which is packaged has intrinsic effect (innate immunity activation)

Is there a size limitation? Answer: No limitation of packaging, but of RNA production up to 50kb done but not fully clear yet if it works

How can you compare the antigen dose between two systems? Answer: Less protein produced from

Liposome particles but compensated by innate response.

Ralf Wagner, Institute of Medical Microbiology & Hygiene, University Regensburg

New Strategies Towards Vaccine Design - Challenging HIV?

- immune response induced by vaccine has to focus on conserved proteins and epitopes to limit immunological escape  knowledge on molecular epidemiology and specific targets necessary

- HIV and human cells have different codon usage  RNA- and codon optimization can increase expression and RNA nuclear export of viral genes in vaccines


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- 1 st generation (EuroVacc): T cell vaccine to induce broad and polyfunctional T cell response that mimics that seen in long-term non progressors (LTNP)

- Vaccine sequence from clade C (CN54) as DNA vaccine (DNA-C) or delivered by New York Vaccinia

Virus (NYVAC-C)

- highest IFNg T cell responders in DNA-C + NYVAC-C group compared to NYVAC-C alone in healthy volunteers, T cells are polyfunctional, more Env- than Gag-specific T cells

- Re-Design I of 1. T cell immunogens, 2. immunogen formulation and 3. poxviral vector (collaborations

CAVD and TVDC)

1. frameshift in Gag introduced for higher expression of Gag in relation to Pol/Nef and production of

Pr55Gag virus like particles (VLP)

2. Gag VLP (DNA vaccine) induce strong anti-Gag T cell response in Balb/C mice but this is reduced when Env is co-administered; spatial separation and ratio modification of the formulation lead to balanced and high level T cell response for Gag, Pol and Env antigens

3. delivery of 2 bivalent pox viruses, one encoding Env and the other one encoding the other HIV genes

- Re-Design II: 1. immunogenicity in non-human primates (NHP), 2. optimization of immunization schedule in Rhesus monkeys

1. replication competent NYVAC-KC and replication deficient NYVAC, in combination with DNA prime and gp120 protein prime with MF59 adjuvant; scarification vs. i.m. for NYVAC-KC

 DNA prime groups induce more polyfunctional CD4 and CD8 T cell responses focused on Gag/Pol; no difference between NYVAC and NYVAC-KC and route of injection

 groups without DNA-prime show higher antibody titers in the early phase of immunization and all groups, with or without DNA prime have similar antibody titers in week 48

- Future plans: o Acceleration of prime/boost immunization schedule of T cell polarizing vaccine regimens o Development of immunization regimens resulting in a synergistic effect between T cell and antibody polarizing vaccine regimens

- Questions: o Will this resemble humans because antibodies in monkeys are always higher? o Are antibodies neutralizing N1 virus? o Did you challenge the animals? No, because there is no Clade C virus for challenge available. o Why didn´t you use adjuvants? DNA and viral vectors are intrinsic adjuvants.

Peter Palese, Head of Microbiology, Mount Sinai School of Medicine

Improving Influenza Vaccines

Good vaccines available

>1 in 5 hospitalizations in the US influenza related (Jan. 2013) because strain was missing in vaccine.


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Vaccine formulations (2000-2010)

All had H1N1 H3N2 and Flu B components; changing strains but often stayed the same, nightmare for vaccine producers. Problem; vaccine was shipped later than epidemic occurred in 2009; big impact on epidemic not possible this way.

How can we do better?

Hemagglutinin Stalk-specific antibodies



Human stalk-specific monoclonal ABs have been identified



We can generate monoclonal abs of such specificities in the mouse

 Nature uses this mechanism to eliminate “earlier” strains- extinction of sH1N1 by pH1N1 viruses

Cross-reactive Abs binds to Stalk region of hemagglutinin.

Strategy for boosting the AB response against the conserved regions of Influenza virus hemagglutinin.

The Stalk is better conserved than the “head” of the protein.

Elimination of seasonal H1N1 viruses by anti-stalk antibodies: pre-2009 is being eliminated, but pandemic H1n1/2009 had different stalk. >Elimination of pre-2009 H1N1.

Patients infected with pH1N1 have higher Titers of Abs that neutralize cHA-Expressing virus.

Production of chimeric Hemagglutinins with specific stalks>Assay for stalk specific antibodies

Summary:

Development of analytical tools to detect and quantify ABs that bind the hemagglutinin protein and neutralize influenza virus but do not have hemagglutination inhibition activity

Influenza virus pH1N1 infected patients have a higher titer of stalk-specific Abs

Induction of protective levels of stalk-reactive Abs using chimeric HA constructs in mice; Challenge with all viruses of H1-subtype: Also protection against H5N1 and H6N1.

Prime boost/ with different HAs, but always same stalk

Mechanism of action of neutralizing anti-stalk antibodies:

Don´t inhibit hemagglutination activity>Hypothesis for mechanism:

Abs are attached to the virus and get endocytosed with it, inhibit fusion and uncoating, Inhibit budding from cell, HA maturation, and help complement binding/activation.

Targeting group 2 HAs (including H7 and H3). H3 Stalk

Prime boost/ with different HAs, but always same stalk.


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Conclusions from proof of principle studies

Broadly protective immune response to the stalk domain can be induced

Chimeric HA constructs protect mice from challenge with heterologous and heterosubtypic virus strains.

The observed protection is antibody mediated

Conclusions

Immunization with chimeric hemagglutinins induces broadly protective ABs directed against the stalk domain

Vaccinated animals are protected against homologous, heterologous as well as heterosubtypic challenge

Questions:

How different have the heads to be/Why don´t we see it in patients? Answer: We see them in old patients! Probably more than 20% difference in head

Have you tried to use head alone? Answer: Head alone not very stable

Infectivity enhancing activity of stalk-ABs? Answer: Hasn´t been observed with Influenza. You don´t see enhancement in patients who have stalk -ABs.

Can virus escape/stalk change? Answer: Possible, hopefully enough fitnesscost so it couldn´t change too much.

Lutz Gissmann, German Cancer Research Center, Heidelberg

Immune response against HPV infections: implications for natural history and vaccination strategies

- HPV lifecycle is restricted to the cervical epithelium, there is no viremia

- the virus is thought to infect the basal cell layer of the epithelium via microabrasions

- new virus particles are assembled in the upper layers of the epithelium and virus is released with the cells as they are shed from the epithelial surface

- cervical cancer is a rare and late consequence of a frequent infection

- HPV related cancer in women 527,100 and men 33,800 cases worldwide/year

- seroconversion after HPV infection is inefficient and slow, titers are low: two years after DNA detection only 75% have weak antibody responses

- role of antibodies in controlling natural HPV infection: o neutralizing antibodies prevent infection in animals o warts are less frequent with older age o protection against new infection depends on high titer of antibodies (Cervarix, PATRICIA HPV-008 study)

- events leading to infection (very long process over days): capsid binds to HSPG on basement membrane (can be blocked by anti-L1 antibodies)  conformational change  accessibility of L2 to


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proprotein convertase (PC) cleavage  17-36 region of L2 is exposed (anti-L2 antibodies)  capsid can be transferred to epithelial cell receptor

- HPV vaccine: 360 copies of L1 protein assemble spontaneously to virus like proteins

- vaccine-induced neutralizing antibody titers are much higher than after natural HPV infection because: o amount of antigen is higher o VLPs display epitopes that are covered in infectious particles o papillomaviruses have mechanisms to escape the immune surveillance and these mechanisms do not apply after systemic vaccination

- vaccine induced antibodies reach the site of infection (detectable in cervicovaginal secretions) and are detectable for at least 9.4 years and until now no infections in vaccinated women from clinical trials

- but: prophylactic vaccines do not cure existing infections; neutralizing antibodies needed to prevent infections and E6/E7-specific cytotoxic T cells to cure infection and disease

Questions:

How many are not responding to vaccine? 1 in 6000.

Best delivery for therapeutic vaccine? Overlapping peptides, study in Netherlands with 67% success in lesions.

Regional differences in HPV subtype distribution? Everywhere 70% are HPV 16 and 18, 3 rd subtype different everywhere.

How do binding and neutralizing titers correlate? Neutralizing titers are 5 times lower, but detection assay is not perfect.

Will vaccine help other strains to come up? Strains spread independent from each other so no niche is set free by vaccine. (remark: echoviruses did not take over poliovirus niche either)

Can we learn from HPV therapeutic vaccine for others? Big pharma companies are not interested

12:15-13:15 Lunch

13:15-15:15 2 nd

Question: How can we achieve sustained antigen responses?

Chair: Ramesh Akkina, Colorado State University; Gerd Sutter, LMU-Munich;

David Topham, University of Rochester;

Immune responses after influenza vaccination

Conclusions:

Prime dose of avian H5 or H7 IAV or LAIV primes for responses to subsequent suboptimal dose of inactivated vaccine

Initial priming occurs despite lack of measurable serum immune responses

Cellular basis of the memory is unknown.


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Immunity to emerging influenza

Pre-pandemic vaccination can prime for improved response to pandemic strain

Minimizes need for two-dose schedule

May provide dose-sparing strategy

Recent advance in immune assays allow early prediction of vaccine success

Optimal T-cell mediated protection may require mucosal tissue involvement

Durability of protective immunity

Durability of tissue resident memory T-cells may be limited

Pre-pandemic vaccination elicits a sub-clinical immune response

Questions:

If resident T-cells are needed, does T-cell response in blood matter at all? How do you activate T-/Bcells at right location? Answer: Mucosal site of immunization/mucosal administration of adjuvants/ Need better assays.

Killed/live vaccine comparison: Which one is more immunogenic?

Systemic AB responses are enough to prevent infection on genital Way, see HepB and HPV. What is different in the respiratory route? Not enough of right Abs at right place>Abs in circulation don´t get to respiratory epithelia?

Christian Kurts, Institute of Molecular Medicine & Experimental Immunology, Bonn

Optimizing CTL induction by chemokine-enhanced cross-priming

- cross presentation: uptake of exogenous antigen and presentation on MHCI molecules

- necessary in vaccinations for inducing a cytotoxic T cell (CTL) response

- TLR ligands (adjuvants) increase signal 2 (co-stimulation signal)

- Mannose receptor bind to antigen  endocytosis  SEE early endosome  MHCI presentation

(different route for MHCII presentation)

- Necessary for cross-priming: o TAP transport to EE, which can be induced by TLR4 ligands o Licensing of DCs by T helper cells

- Licensed DCs produce CCL3/4/5  bind to CXCR5 on CTL  CTL will be attracted

- Alternative pathway of cross-priming involves NKT cells and CCL17 (produced by DCs)

- NKT cells recognize glycolipid antigens presented by DCs on CD1d molecules o Anti-galactosylceramide (a-GC) is a synthetic ligand

- OVA + a-GC increases the ova-specific CTL response compared to OVA prime alone (also in

MHCII-/- mice but not in CD1d -/- mice)

- CCL17 GFP reporter mouse: iNKT cells induce CCL17 production in splenic DCs (CCL17 is normally not expressed in the spleen)


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- The same DC that cross-primes the CTL has to produce CCL17 and CCR4 expression on CTL is required

- A-CG and CpG added to OVA prime lead to a faster clearance of Adeno-OVA-Luc compared to both adjuvants alone

- NKT cell help could be defined as “signal 0” in CTL cross-priming

- Implications for vaccine design: o Chemokine hotspots: CTL can locate relevant DCs faster (relevant= containing antigen and optimally matured) o CCR4 and CCR5 ligands synergize at enhancing NKT and T helper cell licensing and cross-priming of DCs  synergism may be exploited to improve vaccinations, combing adjuvants for T helper and NKT cell induction

Questions:

How many adjuvants would the FDA approve?

Will it work in humans because they have less NKT cells?

Percy Knolle, Institute of Molecular Immunology, Munich / Bonn

Key role of CTL expansion in the liver for successful therapeutic vaccination against chronic viral infection

The liver in immune regulation:



Associated with induction of immune tolerance exemplified by transplantation tolerance

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Rapid and efficient clearance of circulating antigens by hepatic scavenger cells

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Antigen-presentation by hepatic scavenger cells generates regulatory CD4 T cells or liverprimed memory T cells



Immune competence in a non-lymphoid organ

Local regulation of immune responses in the liver: Kupffer cells, T-cells, dendritic cells

Conceptual problem: Inflammation>antigen presentation by mature DCs>immunity>memory

Steady state-no inflammation>antigen presentation by immature DCs>T-cell deletion>hole in T-cell repertoire

Pathogens with innate immune escape, Systemic dissemination of viral proteins in absence of inflammation

Deletion of virus-specific T-cells early in infection

- Liver-primed CTLs are not eliminated;

- generated in the liver under non-inflammatory condition

- Prevent attrition of T-cells specific for circulation antigens

- Complement conventional memory T-cell formation during inflammation

- Protect against viral infection


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- 5-8% of antigen-experienced CTLs in mice and man

- can be employed for therapeutic vaccination

Liver-primed T-cells generate recall responses, liver-primed T-cells are distinct from other T-cells, liverprimed memory T-cells

Liver-primed T-cells are



Liver microenvironment, inhibition of T-cell function and proliferation, successful hepatic immunity; early recognition of pathogens within sinusoids, NK T-cells, granulocytes, Kupffer cells and platelets; relevance shown for bacterial infection



Sufficient numbers of effector T-cells; liver functions like a maze, generation of large numbers of

T-cells, recruitment to the site of infection

CD8-Tecell expansion in the liver facilitated by TLR signaling

Relevance of myeloid cells for local T-cell expansion in the liver;

New structures: iMATEs > T-cells associate within. Proliferation of T-cells happens only in iMATEs for local expansion of T-cells,

Total different structure compared to other immunological structures: no Collagen IV, but gp38 (tissue organizing cells),

Relevance of iMATEs for clearance of viral infection in the liver: example LCMV, appear after d6 and disappear at day 13 when infection is gone

Prevention of iMATE formation leads to failure of CTL-mediated viral clearance. No iMATEs formation in chronic LCMV infection, but you can induce them by injection of TLR9-L.

Relevance of iMATEs in clearance of chronic HBV infection.

AdHBV-Modell, DNA-vaccination is not sufficient to clear infection, but with co-injection of TLR9-L you get expansion of T-cells more than 10x

Summary:

- iMATEs: a novel anatomic structure in the liver facilitating local CTL-expansion

- inflammatory signaling induces iMATE formation

- inflammatory monocytes/dendritic cells form iMATEs

- massive CTL expansion in iMATEs iMATEs facilitate jump-expansion of CTLs initially primed in lymphoid tissues

New model of phases in vaccination:


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Prime-boost vaccination>lymphoid tissues priming of immune response>iMATEs CTL jump expansion

Up to 50-300-fold expansions in iMATEs

Questions: cells originate from bone marrow? Yes!

Could you demonstrate iMATEs formation histological? No good mouse model, not possible in humans.

Question: Worries in humans. Antiviral treatment first to reduce infected hepatocytes, then use TLR9-L

Are patients with cirrhosis in worse position? Yes, and also seems like doesn´t work in livers of older mice. Only in healt hy liver. Probably don´t play a role in liver cirrhosis.

TLR9 recognized by liver or myeloid cells? Not sure.

Damage of liver? Little, not too much.

David Pauza, Institute of Human Virology

T follicular helper cells and the response to HIV vaccines

- HIV Env binds to CCR5 and CD4 on T cells

- In CCR5+ but not in CD4-



T cell binding of Env p38 MAP kinase and hence caspases are activated to induce cell death

- When Env binds to CD4, Akt is activated and p38 is suppressed, which allows cell survival

- Lots of Env is present in germinal centers

- Env signaling through CD4 can promote activation of T follicular helper cells, which are crucial for B cell differentiation and antibody production (could the increased B cell production be the reason for hypergammaglobulinemia in HIV patients?)

- Env signaling might also promote CD4 cell depletion by increasing proliferation of Fas+ Tfh cells, thereby increasing susceptibility to Fas:FasL induced cell death

- An Akt inhibitor could be used to reverse Tfh activation and cell death

15:45-17:15 3 rd

Question: Do we have examples for sustained vaccine responses?

Chair: José Esparza, Bill & Melinda Gates Foundation; Anders Vahnle, Karolinska Institute

Stockholm

Antonella Folgori, Director of Immunology, Okairos, AG

Sustained T cell responses induced by viral vector vaccines: preclinical and clinical evidence

Adenoviral vectors to express antigens.

Problems: Pre-existing immunity to vectors, i.e. Neutralizing Abs are present in many humans. Also true for chimpanzees.

Search for novel adenoviral strains in chimpanzees, isolated and screened samples from over 700 nonhuman primates>identified more than 100 strains grouped into 25 distinct serotypes. These have high immunological potency in macaques, Macaques immunized with different Adenov. Expressing HIV

Gag.


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This T-cell response is sustained for years (50% positive cells in ELISpot 6 years after vaccination).

After 2 nd Adeno immunization these expand again.

ChAd3 provides full protection against Ebola challenge. Phase I trials with HCV, Malaria and HIV vaccines of different Serotypes (ChAd3 and ChAd63)

ChAd3 induces poly-functional T-Cells in humans as shown in HCV vaccine program against NS3-4a.

Prime/boost strategy with different group C Ad vectors in NHP and in Humans: Prime ChAd3 prime,

Ad6 boost, Ad6 prime ChAd3 boost). Successful in NHP; But not as efficient in humans. Here ChAd3 is the better primer. No correlation with anti-vector T-cell response, some correlation with anti-vector nAb

>Change to MVA as vector:

Adenov Prime, 8 weeks later MVA boost:

In NHP, very good response

In Humans almost the same!

Holds true for HCV, HIV and Malaria vaccine trails.

ChAd prime/MVA boost induces broad T-cell response. Cross reactive across divergent HCV genotypes.

This T cell response is well above the protective threshold (as published by Spada et al, 2004, Gut,;

Folgori et al, 2006, Gut): 280 SFC/10e6 PBMC

>Also induction of high levels of memory cells in humans

Also induction of good antibody responses in humans.

Now FDA approved Phase II efficacy study in high risk individuals.

Malaria vaccine: long-lasting immunity: after 2 years still high number of T-cells, also after readministration of either vector there is a rapid expansion.

The invariant chain (li) is a candidate enhancer of T cell response



Screened several genetic adjuvants for their ability to increase immune response in mice using

Adeno vectors



Invariant chain is a protein involved in the formation and cell surface display of the MHC-II complex



Li was shown to increase presentation on MHC-I

Fusion of MHC-II Invariant chain increases CD8 T-Cell response in mice> ChAd3-Li

Questions:


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Side effects: Enhancement of HIV infection after Adenovirus vaccine: Strong difference between Adv-

Genotypes. Protection in Ebola study is mediated by CMI.

Why were only non-structural proteins used in HCV trials. Answer: Thought to be better for broad response.

HCV-vaccine tested in NHPs?

Are Strains of ChAd oncogenic in other animals, e.g. hamsters? Not tested, but nothing observed, they are not replication competent.

Yiming Shao, China CDC

Experiences from the 1st lentivirus vaccine, equine infectious anemia virus

- 1905 EIAV (Equine Infectious Anemia Virus) discovered, lentivirus, epidemic in animals

- Mid 70s: EIAV vaccine Dr. Shens group in Chinese Academy of Agriculture

- 2002: FIV vaccine 2 nd lentivirus vaccine

- Envelope gp90, gp45 transmembrane proteins

- Acute phase: 3 months, chronic phase 9 months; then unapparent, fluctuating rectal temperature (>39°C)

- LN40 very pathogenic strain, passaged trough horses and donkeys, donkey leukocytes and fetal donkey dermal cells to obtain live attenuated vaccine

- after large scale vaccinations from 1978 on EIAV infection rate dropped from approximately 5 to

0 % in 7 years (likewise horse mortality and rate of slaughtered horses decreased)

- What happened through passaging?

- Critical mutations in envelope, glycosylation sites are lost, casein kinase II phosphorylation sites are gained

- Generation of infectious recombinant clone from vaccine  protection only partial

- CD4 and CD8 IFN-g and proliferation is higher in protected than infected animals

- Until 6 months only partial protection against challenge, high titers of neutralizing antibodies come up later and peak at month 6

- EIAV vaccine as live attenuated viral vector for HIV

- Introduce aa differences between HIV and EIAV envelope in viral vectors

- Rhesus macaques + DNA rTV vaccine: protected against homologous challenge, but not in heterologous challenge sHIV-162 P4

- Group C: Vac + TDF/FTC strong decrease in viral load

- China CDC HIV vaccine clinical trial: induction of HIV-specific T cells and antibodies, starting recruitment for phase IIb in 2014

Questions:

Does vaccine virus persist? Yes  somatic hypermutations. Mutations of vaccine virus can occur.

Can virus from vaccinated horses spread? Not assumed, because viral titer is so low.

Vaccine also protects against European strains.

Same effect in newborn horses? Not known, vaccine given at age of 3 months.


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Gallo comment: Attenuated SIV has been a disaster in primates, killing young animals immediately and old animals eventually.

Which cells are infected? Virus propagates in horse PBMCs.

Luigi Buonaguro, National Cancer Institute, Italy

VLPs and particle vaccines

Revision of VLP and particle vaccine field:

20 th century vaccines:

- Killed vaccines

- Live attenuated vaccines

- Viral vectors/inactivated virus better than other methods but are less safe

- Induction of innate immunity is reason why virus vaccines are so good> Innate immunity must be stimulated by any vaccination method to get a durable immune response.

- TLRs play a big role in B-Cell functions.

Advantages of a vaccine approach based on VLPs or particles:

- Highly repetitive surface

- Display of foreign epitopes

- Co-delivery of antigen and activators of innate immune system

Core protein of virus expressed in cell> antigens assemble to make particles which are non-infectious.

Examples of enveloped vs. non-enveloped VLPs (HIV vs. HBV)

Virus-like particles as vaccines: HBV and HPV

To types of attaching antigens to particles: Passively targeted (A) and actively-targeted particles (B)

Co-adjuvants can be incorporated in delivery systems>various methods and candidates available

Cross-presentation for exogenous antigens delivered as particles important:

In this way activate CD4 and CD8 T-cells

TLRs (3 and 8) have been shown to be crucial for Maturation of MDDCs.

Many Virus-like particles are currently in vaccine development

- HBV and HPV licensed.

- Most clinical trials are on HPV vaccine

- Immune response to HPV-/HBV- VLP vaccines is long-lasting.

- HBV-VLP can be used to deliver other antigens: long list of candidates.

- Influenza vaccine based on VLP has been developed: Mostly expressing M2-protein

- Influenza nanoparticle vaccine: HA and Ferritin forming particle

- Norovirus VLPs induce partial protection


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- HIV-VLPs induce a long-lasting memory in NHP, low titer, but good response after boost>good memory

- HIVp24 nanoparticles for presenting conformational HIV Env Antigens in development; No

Membrane needed.

Synthetic vs. biological systems>the two systems get closer to each other recently

Conclusions:

- Great opportunity to design advanced platforms to simulate APCS, especially DCs, combining and delivering the target antigen and if needed an immunmodulator

- This safe strategy shows improved immunogenicity and induction of prolonged immunological memory

- Several pre-clinical and human clinical trials, showing very promising results, ensure a significant pipeline, which likely will provide results to move forward to licensing and marketing of vaccines based on this technology

- In Particular, human clinical trials performed using enveloped VLPs for Influenza, RSV,

Rotavirus will provide essential information on the manufacturing optimization, safety and immunogenicity on VLP-based vaccines.

Demands in the developing world

Lan Anh Nguyen, Natl Inst of Hygiene and Epidemiology, Vietnam;

- National and Regional Institutes responsible for training and vaccines, EPI

- Vaccines introduced in Vietnam: o 1985 Tuberculosis, Tetanus (pregnant high risk, 1.6 million), Polio, Diphtheria, Pertussis,

Measles o 1997 HBV, Japanese Encephalitis (high risk, 2.5 million), Cholera, Typhoid (only in high risk areas) o 2010 Haemophilus Influenza B in DTP-HBV-HIB combi o planned: Rubella, Rota, Influenza

- Funding: 50-70% government (material, training), 5-10% local (Communication on vaccination campaign), 30-35% International agencies (Technical support, vaccine supply, cold chain)

- Polio eliminated 2000 (risk of wild polio intrusion from other countries), maternal and neonatal tetanus

2005, 2003: HBV vaccine for everyone, 2010 HBsAg prevalence is 2% in children under 5 years

- Mortality rate of children under 1 year, from 2,3% in 1990 to 1,2% in 2010

- Improve immunization rates in remote areas for infants under 1 yr with immunization coverage lower than 90%

Ramou Njie, The Gambia, Gambia Hepatitis Intervention Study

- hepatologist, running Gambia HBV intervention program

- 1986: 60,000 children received HBV vaccination

- 1991: available to every child


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- same time cancer register established, 80% of liver cancers attributed to HBV (most common cancer in young men 20-40y)

- CHB patients with decompensated liver disease are discriminated because they cannot ask for treatment funding, only when co-infected with HIV

- Community based screening for HBV  included in EU-funded study

- Patients often die within 3 months after diagnosis

- Suggestion: contact NIH institute for cancer because of program for help with non-infectious disease in developing countries

- Not so much perinatal transmission but early in life, reason? Food sharing? Mother chewing food for child?

Perspectives from Funding Agencies

Guntram Bauer, Human Frontiers Science Program;

Funded 25 years ago by Prime Minister of Japan Mr. Nakasone to “Bring scientists of different disciplines all around the world together”

1990 First grants and fellowships

Member countries:

Canada, France, Germany, Italy, Japan, UK, USA, EU, Switzerland, Australia, S. Korea, New Zealand,

India, Norway

You can get grant if your laboratory is in one of these countries. Or recruit someone from these countries

Annual budget: $56m US Dollar

HFSP support emphasizes:

- International and intercontinental scientific collaborations

- Novel, innovative and interdisciplinary approaches to basic research in the life sciences, not asking for preliminary data

- Strong emphasis on participation of scientists from disciplines outside the traditional life sciences

- Bottom-up investigator driven: no priority areas

- Scientists who are early in their careers

Funding mechanisms:

Research abroad>Establish laboratory>research collaboration

- Postdoc fellowships (60-80 a year)

- Transition to scientific independence (group leaders)


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- Intercontinental teams (group leaders and senior scientists) (2/3rd of budget)

Program Grants:

For scientists at any stage of their careers (emphasis on young team member)

Young investigator grant

All team members within 5 years of obtaining an independent position. Not more than 10 years after the

Ph.D.

Deadline:

Letters of intention: Mid-March

Full applications: early September

Trimmed down to 250>90 are invited for full applications>30-35 funded

Objectives:

To stimulate novel, daring ideas and innovative approaches, preliminary results are not required in research grant applications

A clear preference for participation from scientists outside the life sciences to understand the complex structures and networks that characterize the living state

Applicants are expected to develop new lines of research through the collaboration

Team:

PI must have lab in a member country

International/intercontinental composition

Individual team members come from different scientific areas (life sciences, physics, and chemistry)

Budget:

(In US Dollars)

250,000 per year for a team of 2

350,000 for a team of 3

450,000 for a team of 4 or more (maximum funds)

Three year grant

Success rate: 5-6 % (ca. 33 awards p.a.)

More information: www.hfsp.org

Friday, May 31, 2013

Global Virus Network Semi-Annual Meeting

Seminar Room, Institute of Virology, Technische Universität München, Trogerstr. 30, 81675 Munich


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Sharon H. Hrynkow, GVN President

Welcomes everyone. Where is GVN at this time point? What are the next steps? Much has changed since the last GVN meeting. The GVN is all about science.

Robert C. Gallo, GVN Co-founder and Chair, GVN Scientific Leadership Board

Welcomes two new centers: Brazil and Univ. Rochester. Interest to expand to England.

Welcome Notes from the German GVN Branch

Reinhard Burger, President, Robert Koch Institute

Last months showed how important such a network is with the spreading of H7N9 and the new corona virus. This shows how fast viruses can spread and how difficult to detect. No new cases of H7N9 in last weeks. 6-7 Billion US Dollars lost from markets, farmers closing markets and culling of chickens.

The new coronavirus is also special:



Contagious (but not very)



4-5 Transmission to health care workers

 Classified in German newspaper “virus of rich old men”



The information flow is so far poor



Exchange of samples, control etc. is not good yet



Has been mentioned at recent WHO general assembly as a worrying threat

Thomas Mertens, President, German Society for Virology

Introduces the Society for Virology:



1050 members



36% female



64% male



180 Students, gender balance reverse: 59% females; 41% males

Separated from Microbiology Society in 1990, Includes Germany, Austria and Switzerland

 medical faculties: o 36 in Germany o 4 in Austria o 3 in Switzerland

 other institutions (e.g. MPI, Helmholtz, Leibnitz, RKI, FLI)



52 Chairs



86 Professorships

Second society in Germany: DVV (Gesellschaft für Virologie): No individual members

What could GVN and Society for Virology do together?


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Ulrike Protzer, German Center for Infection Research

Introduction of the German Center for Infection Research:

Founded only 2 years ago.

One of 7 centers of health research of Fed. Gov. of Germany.

For all >200 Million Euro/year for health research

>Dedicated to global disease burdens including cardiovascular, lung, diabetes etc.

Set up as competitive contest

Includes different institutions doing infection research (University, Research Institutions, Military etc.)

Introduction of GVN Board Chair by Anthony J. Cernera, GVN Secretary and Founding

Board Member

Reminded that GVN is relatively new organization:

- January 2011 the GVN idea was born

- 2 months later 25 scientific directors meet in Washington DC

- Has grown a lot since then

- Board has fiduciary responsibilities, consistent with non-profit boards based in the US

Introduces GVN’s 2 nd board chair: Kathleen Kennedy Townsend, former Lt. Governor in Maryland, oldest child of US Senator and Attorney General Robert Kennedy.

Remarks from the GVN Board Chair

Kathleen Kennedy Townsend

What was going on last year? What are the questions we are dealing with? What´s your vision for how to go forward? We have to go together.

Stresses progress within 2 years: Board members from 5 members to 25 members. If you grow too quickly it´s a challenge to build together.

2 nd question: What are we supposed to do? How should we do it? Best scientists from around the world: share ideas and work together. But what are the criteria to be one of the top scientists? How do you measure who is the best? How do you politically organize so the whole world is included to prevent resentments.

What is your responsibility to bring in other groups. How do we include the developing world? If we only include the developed world we will not get approval from developing world.

From the Board perspective, what about Fundraising?

How are we going to do it? Big challenge. Conferences around the world. Raising money for conferences is doable. But it ´s not how we are going to be measured: We need to Build / train scientific capacity / exchange programs.


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Best way to raise funds is to through building scientific capacity. Everyone knows members/leaders can raise research money. How do we fight epidemics, how do we built the capacities?

Plenary: “The State of Healthcare Worldwide: A Rallying Call for GVN”

G. Steven Burrill, CEO, Burrill & Company, GVN Board Member

- Sometimes you get lost in the world of science and don’t see what is around you

- World of health care is much more complex

- Without energy, food and clean water nothing in health care is relevant

- In healthcare: Procedures  outcomes, cost  value

- Healthcare systems face chronic problems, e.g. population aging and falling birthrates: the ratio of working people paying into healthcare system to those using the public benefits shrinks

- Annual costs of chronic disease in the US in USD B (75% of total costs!): o Heart disease and stroke = $653 o Cancer = $396 o Diabetes = $226 o Obesity = $209 o Arthritis = $190

- Patient, doctor and payer are disconnected, no one knows what costs what

- Payers (government, health insurance) pay for outcome of value

- On average, 60% or 3 trillion dollars spent on healthcare during the last year of life

- Keeping everybody alive costs a lot and will bankrupt the health care systems

- Non-communicable diseases take a growing proportion of health care costs

- Urbanization and technology is changing (data connect to doctors, home use diagnostic tests)

- Burden of healthcare: 11% of cost are drugs, 37% hospitalization

- Pharmaceutical industry least respected, less then tobacco industry

- Not enough doctors to cover people

- What is my personal burden to stay well? People do not pay for their own healthcare  there is no penalty to be sick, system would be different if you had a charge (individual responsibility)

- Should safety and efficacy alone be the standard for drug approval, or should cost be a factor?

- Value capture outside the product (e.g. Facebook is "free" but hi value) for health care?

- NIH grants for disease research in 2012: 10,302 million (7%) virus-related 143,000 million other health issues.

- 3 areas to combat viruses: o improve health (nutrition, sanitation) o prevent infection (improve surveillance and reporting, advance technologies, production of vaccines) o treat infection (therapeutic vaccines, antiviral drugs, gene therapy)

- Google flu trends to predict outbreaks, how many Google for same symptoms at one time point?


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- Personalized medicine: know molecular mechanism of disease to treat the disease instead of the symptoms

- Concerted effort needed to solve the problem of dramatic viral risk to global population: o Global health community and governments must work together o Create supportive health systems and infrastructure o Reach remote and marginalized populations o Develop new and improved technologies and vaccines o Stimulate PhDs to enter the field o Improve networking of anti-viral researchers

Questions / comments:

60% of healthcare expenses used to keep old people as long alive as possible. There should be a change because the reason is mainly cultural. Not the peoples demand or in their interest.

Big problems are very local, e.g. local health care system. How can we solve problems globally then? All around the world three different levels of health care: Government level, some privatization (private health care), private (rich can pay for everything).

Human nature, they only learn from their own mistakes. We have to share mistakes, but this seems to be against human nature.

What do you see as a vision for GVN? A.: 1. Keep research funded and focus on it. 2. Public information>world must understand

“outbreaks”. 3. Help link people around the world / Network.

Report from the GVN President Sharon H. Hrynkow

Goal of the presentation is to provide a status report on GVN progress since the last meeting in

October, and to share info on where GVN is planning to go in the coming months. We are making steady progress. Hrynkow asks for input from Center Directors: GVN is a coalition of institutions, all of which should be involved in creating programs, initiatives.

GVN milestones since October

Organizational strengthening:

- Board of Directors expansion, more global, more diverse

- Scientific leadership board

Operational strengthening:

- 501c3 in April 2013, tax free in US, milestone

- Business plan adopted Feb 5th

- Working towards sustainable funding

Graphic developed to help explain GVN to range of audiences -- The GVN “house”:

3 major areas pillars of activity:

- Research & Training, go hand in hand

- Education of public, credible, authoritative information on outbreaks and related research

- Advocacy to governments and private groups for support on issues and budgets


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Basement and ground floor = Global Collaboration, Capacity Building and Top Science . “Roof” represents the GVN goals of improving health and having better pandemic preparedness.

In more detail:

Research and Training - pillar:

Network meetings advance research agenda

- Munich

- Moscow, September 2013

- Xi ´an, China, Spring 2014

New Centers of Excellence joining network

-Rochester NY and Brazil

Rapid response fund-Opportunity Grants

Useful tool to respond fast

- Small grants to GVN teams plus others

- Quickly reviewed

- Push new lines forward to create pilot data

- Expected to announce full program in July

Post-Doctoral Training program

Broad based skill building with opportunities to trains at more than one GVN center of Excellence

Near term goal

Short term exchange programs

Rina Shah “STEP” program

Exchange across the network for skill building, collaborations

Robert Karrs, Program director for STEP and thanks to 3 peer reviewers (Salvato, Menendez-Arias,

Brack-Werner)

Rina Shah STEP awardees

Dr Aysen Gargili (UTMB-Colo. State) - Heartland virus

Dr Marit Kramski (U. Melbourne to Scripps) - HIV

Dr Runqing Jia (Beijing Inst. Techn. To IHV) - Monoclonal Ab techniques

Model training programs at GVN Centers- examples

- Masters level Training: GVN Scholars at Billy Hall's Ireland Vietnam Blood borne Virus Initiative

(IVVI)- degree program

- Short term training opportunities: 3 PhD and Masters student

Education pillar – website is primary tool for education efforts right now.

New Webpage design and website partners presented


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- UST-Global, early conceptual partner

- CTIS, Inc. Ongoing support

- Dr. Florence Haseltine, w omen’s health leader and IT expert, now lending service to GVN

- Nora Grannell, lead for GVN on communications

GVN is using twitter now > start to use social media. Follow us @GlobalVirusNews

Public Education/website project:

Testimonial Project

Challenge: Viral disease has many faces: young, old, rich, poor, diversity of ethnic and social factors.

How can GVN connect to the public with so many different “faces” for our problem?

The Testimonial Project tells the personal stories of people affected by viral disease and those working to unlock the mysteries of viruses in order to prevent infections and treat disease

Stories on website to use personal stories to communicate information:

- Dr. Stephan Becker, Marburg University: Stigma, duty for informing the public about infectious disease

- Dr Salim Abdool Karim, South Africa Medical Research Council: Epidemiology of AIDS, especially vulnerability of young women to infection related cultural norms

- Dr Barry Beaty, College of Vet. Medicine, Colorado State Univ.: focus on Dengue

Will be expanded to include personal accounts of other centers and patient stories. Thanks to Roxanne

Bauer, graduate student intern, Syracuse U./Maxwell School for great work on this project.

To Link to the first person accounts: Web-based educational tools

Breaking News status on outbreaks

Advocacy pillar

- Advocating for budgets for medical virology and action

- Example: Scientific Declaration to eradicate Polio. GVN as an organization, plus all three cofounders in their own institutional capacities, called for last mile research efforts.

Financial plans as reported in Business Plan

Funding:

Target: In 2015 generating 15 Million US Dollars

Partnerships with GVN on the rise; GVN seeking to partner with Brazil ’s Science Without Borders program; GVN and The Academy of Sciences for the Developin g World signed agreement…


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Individual country support: e.g. China, granted $50m to its GVN institutions to promote their work in medical virology

Facilities:

GVN moved administrative offices to U. Maryland BioPark: a stone´s throw from IHV, proximity to Dr.

Gallo

GVN Team Members

- Robert Karrs: Program Director

- Nora Grannell: Communications

- Maria Salvato PhD: Exec Sec to SLB

- Florence Haseltine MD PhD: Senior Advisor for IT

- Roxanne Bauer, intern

- Legal – outsourcing, pro bono through Greenberg Traurig

- Accounting – outsourcing, partially pro bono

- Development – still need help

Grateful to Board for all its support since October, and glad to hear from SLB and all sources as GVN creates infrastructure and programs for maximum impact on global health.

New GVN Center Presentation: University of Rochester

David Topham, University of Rochester

- John Treanor (director), Steve Dewhurst, David Topham (co-directors)

- In floor hospital rooms next to lab

- Virology faculty: 8 groups (PIs), Viral immunology faculty: 7 groups, Viral RNA biology faculty: 6 groups, Clinical virology faculty: 6 groups

- Multidisciplinary environment, FEVURS, evaluation of vaccines, RPRC Respiratory pathogens research center, HSCCI Health sciences center for computational innovation, CFAR Center for aids research

- CFAR: 1. Working group: CNS reservoirs of infection and aging, 2. WG: computational approaches to understand HIV RNA structure

- New tools: single-cycle infection systems (Luis Martinez) with reporter gene expression

- More training on computational biology needed to stay competitive

- RPRC key facilities: Leadership and admin, subject enrollment and sample collection, data management, experiments and analysis

- NYICE: New York Influenza Center of Excellence, 6 key projects on T cell immunology and molecular biology in influenza infection

- Prediction of influenza and respiratory disease outbreaks by following tweets

- For GVN (mainly respiratory disease): Training and fellowships, infrastructure to conduct clinical studies, SOPs, computational biology


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- For what is funding lacking? Support more fellowships, characterize lung microbiome and virome, public map of RNA secondary structure of emerging influenza viruses, production of

GMP virus, collaborations in virus surveillance

New GVN Center Presentation: Brazil

Myrna Bonaldo and Claudia Nunes Duarte dos Santos, FIOCRUZ, Rio de Janeiro

History:

1900: Founding of the Serotherapy Institute of Rio de Janeiro in May 25 th

First head: Oswaldo Cruz

1908 Renamed Instituto Oswaldo Cruz

Biggest public health institute in Brazil

Oswaldo Cruz Foundation (FIOCRUZ) is a public, non-profit organization: 1900-2013

Main figures:

- 11,000 employees

- 990 PhD, 1100MSc, 1500 post graduated

- 61 reference laboratories for epidemiological surveillance

- 200 annual courses (20 PHD, 20 Master, 10 Professional Master)

- $300M federal budget/year

Organisation:

Head unit in Rio de Janeiro

8 Regional Units

3 more future Units planned

International cooperations all over the world

- Fortifying the NHS

- Research, development and production to meet global health needs

- Participation in policy decisions borders of international organizations

Brazilian Global Virus Network at Fiocruz

7 centers comprising many laboratories

- Instituto Oswaldo Cruz (Rio de Janeiro): 18 Laboratories; 8 National Reference Centers, BSL3 facilities

- Fiocruz Minas Gerais – Centro de Pesquisas Rene Rachou: 4 Laboratories, BSL3 facilities

- Fiocruz Parana- Instituto Carlos Chagas: 1 Laboratory, regional reference center for

Hantaviruses, BSL3 facilities


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- Fiocruz Pernambuco - Centro de Pesquisas Aggeu Magalhaes: 2 Labs, BSL3 facilities

- Fiocruz Bahia- Centro de Pesquisas Goncalo Moniz: 4 Labs, BSL3

- Fiocruz Rondonia - Instituto de Pesquisas em Patologias Tropicais de Rondonia (Ipepatro): 4

Labs.

Students form other GVN centers can participate in training courses as well as in Master- and PhD programs a t Fiocruz as well as the “Brazil without borders” program: opportunity for post-doc fellowships

Jose Esparza, Gates Foundation

Gates Foundation:

- Family foundation, different to NIH etc.

- Gates define priorities: What are areas that create inequities? Lack of health and access to education.

Characteristics of diseases to be funded:

- Important global health problems

- In higher proportionally affect poor people

- Focus on technological solutions

$1.5 Billion /year on Health

$1.5 Billion/year on global development: Education, micro financing etc.

Maria Salvato, Inst of Human Virology, Univ. of Maryland, USA, Recap of the Durability of

Vaccines Workshop from day prior

At the October 2012 GVN meeting in Baltimore, GVN held a panel discussion on Durability of Immune

Response, run by Reinhard Kurth. With 2 other speakers, Dr. Gallo and Dr. Saphire, the GVN recognized that this was a severe problem for HIV vaccines, Ebola vaccines, and some chronic infections, and more research is needed on the development of B and T cell immunity. GVN agreed to convene a full-day workshop to explore specific issues re: durability, and this was held as a premeeting to the Munich GVN meeting. Salvato provided a summary of the Munich Workshop on

Durability of Immune Response to Vaccines, which will be published in The Viruses (MDPI, Basel).

Her report prompted general discussion on problems of health care funding in developing countries, which focus on certain diseases (especially HIV). Patient story from Gambia where patients wish to have AIDS to gain access to health care (and are turned down if HIV negative). How can structures of donors driven, vertical health care system be changed to strengthen the public health care system?

Kathleen Kennedy Townsend asked, if GVN should write a letter, send it to whom? Discussion of way of communication: On website? Letter? Via local media in each country? Outcome: Group should form to produce a letter today and present to the group tomorrow.


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Panel on Strengthening the Network: Next Steps

Framing the challenges and possible solutions- GVN President Sharon Hrynkow leads

Where should GVN go next? What are the critical operational challenges that need attention if GVN is to have greatest impact? One area of pursuit should be creation of a tool that would facilitate sharing of biological samples across international borders. If GVN had such a mechanism in place before outbreaks occur, scientists would be better prepared to share in the eventuality of outbreaks.

Presentation 1. Leveraging resources: The TWAS-GVN collaboration- Romain Murenzi, GVN

Board member and executive director of TWAS (The World Academy of Sciences)

- TWAS origins: Abdul Salam: A dream of science in the south, building science capacities in the south, south-south and south-north collaborations

- 81 countries lag behind in sciences, less than 1 % of global science output

- TWAS (The World Academy of Sciences): 1071 members in 92 countries, 929 fellows in 75 countries in the South, 142 associate fellows in 17 countries

- Headquarters: Trieste, regional offices in Beijing, Bangalore, Alexandria, Nairobi, Rio de Janeiro

- TWAS offers PhD training, researcher mobility, research support, honoring excellence to increase scientific capacity and excellence in the south

- Funding: Governments, S&T civil society, bilateral agencies, multilateral, private sector

- US PhD $200-250,000 for 4 years, 313 PhD fellowships, 141 Postdoctoral fellowships, 48 visiting scientist, 10 for research and advanced training.

GVN and TWAS signed a Letter of Intent so that new forms of collaboration and leveraging of resources could be identified.

Presentation 2. Toward Universal Materials Transfer -- Mark Kaplan, Univ of Michigan (10 min)

- Problems:

- Permission and visas from country with an epidemic to transfer materials

- Permission from a country accepting material

- Permission for intellectual rights from both institutions

- Compliance with international standards of research

- Jealousies of discovery

- Concern about hurting business in the country of origin of epidemics

- Risks to investigators of getting sick from working in epidemic area (maybe cheaper than transporting material)

- Diseases that might be viral (Kawasakis disease, MS, Rheumatoid Arthritis, Hemophagocytic syndrome, lymphoma, ALS, Sarcoid, Lupus) how to get funding for identifying virus?

- Support connectivity for people, who see “weird” diseases

- What could GVN do? o Support travel and lodging for experts


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o Rapid approval plans o Upfront permission to accept and or transfer material o Agreement upfront for intellectual transfer

Presentation 3. Public Relations: Next steps in development and implementation of a GVN

Public Relations strategy – Mathew Evins, GVN Founding Board Member and Treasurer

- Awareness of the problem & the need. Establish why we are here.

- Familiarity with the role & the resources of the GVN

- With whom is the GVN communicating? Communicating with scientists is easy but we have to communicate with the public

- How do we find success? Money, media resource and media coverage: media has to cover what the GVN does and what the centers are, public advocacy

- Encourage China to share samples with Japan

- How do we tell our story, to whom do we tell our story? Who is telling? Tell Nora, what are you working on, what are the stories you want to tell?

- Your research might (not) be a page-turner

- If we are leaders, we have to comment early on important events and not 4 years later.

- Potential Tools to develop: Alerts (example clock on time square for people dying for lung cancer) , communiqués, metrics, newsletter, timely/topical context and perspective (prevent from misinformation on viruses), state of threats – clear & present danger level

- Appeal to people with real pictures and videos

Presentation 4. GVN secure website - Florence Haseltine

Log In: GVN2013

Password GVN2013

Login in to this site in order to comment on the meeting: what worked, what didn’t. We want to know.

- Web site’s Public Face: o Current News and Video o Keeping abreast of latest communications and rapid news update

- Center of Excellence Section-Membership Section (not public, password protected) o Sharing News o Shared technique and tools o Lab to Lab secure site o Training and educational materials o Opportunity grants for GVN Centers of Excellence o Social communications and Networking

 Twitter

 Facebook

 Public face of each center


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o Internal Sitecenter´s members section o Section for each of the centers of excellence to present their programs to other GVN members o Ability to share pre-publication information o Access to GVN meeting documents

 Content o Each center descriptions /and graphics o Text based documents o Educational and reference documents o Any videos describing your work and other interesting stories

Discussion: Due to time constraints, discussion from this session was postponed until the next day. At that time, there was clear support to go forward with development of a universal Materials Transfer

Agreement, increased support for leveraging of funds from other agencies to meet GVN goals, enthusiasm for the directions of both the public and private websites, and support for completion of a

PR strategy that made GVN as consequential as possible.

GVN / DZIF Scientific Highlights (this session was open to the public)

GVN in Collaboration with the German Center for Infection Research (DZIF)

Control of Chronic Viral Infections

 Moderators – Reinhard Burger, Robert Koch Institute

Epstein-Barr virus and latent infection of B cells: a story of epigenetic gene regulation

Wolfgang Hammerschmidt, Helmholtz Zentrum München

EBV is one out of 8 Herpes Viruses

Most humans are infected

First human Tumor Virus

Causes several diseases: Examples:

Nasopharyngeal carcinoma

Burkitt´s Lymphoma

Hodgkin´s Lymphoma

Gastric Carcinoma

EBV is a paradigmatic model to study viral latency

Human B lymphocytes are EBV´s preferred target cells

Biphasic life style:


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1. establishment of latent phase> only genes important for genome maintenance and cell survival are expressed

2. Lytic reactivation>orchestrated by transcription factor BZLF1>lytic phase>Genes for lytic DNA replication an structural components are expressed

BZLF1>molecular switch gene, which induces the lytic phase of EBV in latently infected B cells

The BZLF1 protein Zta is a sequence-specific transcriptional activator

Zta can bind its cognate sequence even if methylated

Key findings

 BZLF1 can induce virus synthesis only when key promoters of lytic genes are CpG-methylated

 BZLF1 critically supports transformation of resting B cells upon infection

 BZLF1 might transactivate cellular promoters to drive quiescent, resting B cells, which are in G0 into the cell cycle- similar to AP-1

 BZLF1 and AP-1 family members are related proteins

 BZLF1 has dual functions. The BZLF1 gene product act at the level of the virus and at the level of the cell

 Its functions depend on CpG-methlyation.

 Depending on the CpG-Methylation status of EBV Genomic DNA latency and virus production vary cyclically

 As a consequence of this peculiar mode of epigenetic gene regulation, EBVs life cycle is biphasic

Questions: Can you prevent reactivation by methylating drugs? Not known

Questions: Sub Saharan Africa >

Burkitt´s Lymphoma > are they exposed to certain toxins/chemicals to cause de-methylation? Answers: Not known, chronic inflammation like malaria

How HIV can undermine antiviral immune response

Frank Kirchhoff, Ulm University Medical Center

- HIV-1 is the result of a recombination event and multiple cross-species-transmission monkeychimp-gorilla-man

- 4 independent transmissions of SIVcpz or SIVgor to man, but only HIV-1M, one strain caused the pandemic. Why?

- Human innate immunity to HIV: TRIM5alpha, APOBEC3G, Tetherin

- HIV and SIV contain viral accessory genes to counteract host immune defense

- HIV-1 vpu gene, HIV-2 vpx gene, some SIV strains do not have a homologue

- Vif  Apobec3G, Vpu  tetherin and CD4 (not antagonizing in HIV-N/O/P strains)

- Tetherin interacts with viral membrane in general, not virus specific


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- Only HIV-1 vpu is optimally adapted to humans, vpu antagonizes tetherin, which blocks virus release and induces CD4 degradation

- In SIVcpz and SIVgor Nef is binding to Tetherin

- One recently discovered HIV-N strain from Togo encodes a Vpu which is highly active against

Tetherin (virus increases fitness)

- Infection of chimp with HIV-1  Nef regains function against tetherin (2 aa in C-loop responsible)

- Human tetherin resistant to Nef? No, not in general, tested with HIV-O

- Nef  NFkB  binds to viral LTR promoter to induce viral gene expression  vpu is expressed  counteracts NFkB (to repress cellular innate immune response)

Questions:

Why did HIV regain nef function in chimps? HIV lost nef function in humans because human tetherin has a deletion so that Nef is not binding anymore. In chimp functioning again.

Do you see tetherin effect in other cells, e.g. DCs or astrocytes? We did only look in T cells.

CCR5 usage is also acquired in HIV not in SIV? Also binding to CCR5 but less specific?

Pathogenicity of HIV-1 M and N the same, so tetherin also involved in establishment of infection?

Maybe Vpu effect on virus release.

Factors restricting HIV infection

Oliver Keppler, University of Heidelburg

CD4 T-cells and their role in HIV Pathogenesis and latent reservoir

Activated CD4 T-cell

- HIV-permissive

- Major productively infected cell type in vivo

- Cell death

Resting CD4 T-cells

- Major fraction of CD4 –cells in vivo

- Largely non-permissive

- Major latent reservoir in vivo (1:10e6 cells)

- Massive cell death in lymphatic tissue

HIV target cells:

- Macrophages

- Dendritic cells

- Activate CD4 T-cells


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- Resting CD4 T-cells

Why are resting CD4 T-cells largely resistant to HIV-1 infection?

Post-entry restriction at the of late RT

Vpx

- Accessory protein of SIV and HIV-2 but not of HIV-1

- Incorporated into virions via a Gag p6 interaction motif (DPAVLL)

- Overcomes a restriction to HIV-1/HIV-2 infection in resting CD4 T-cells

SAMHD1

- sterile alpha motif (SAM) and HD domain 1

- Restricts HIV-1 RT in myeloid cells

- dGTP-regulated deoxynucleoside triphosphate triphosphohydrolase

- tetramerization and phosphorylation important for biological function

- targeted for degradation by Vpx

Blood of Aicardi-Goutieres Syndrome (AGS) patient: Deficiency of SAMHD1

CD4 T-cells challenged>infection of resting CD4 T-cells (in contrast to blood from healthy donor)

The SAMHD1 restriction is beneficial for the virus to prevent sensing in DCs and prevent proinflammatory cytokine production

Q. Is there a direct antiviral effect of SAMHD1? Possibly.

Q. What´s connection between in-vitro and in-vivo? Sensing after infection is crucial. But it´s not possible at this time to answer.

Emerging Viral Infections

Moderator – Warner Greene, Gladstone Institutes

New Generation Humanized Mice: Applications for HIV and Dengue Pathogenesis and Therapies

Ramesh Akkina, Colorado State University

- Humanized Rag2 -/-

 c-/-, sublethal irradiation, transfer of CD34 + stem cells – 8-12 weeks granulocytes, lymphocytes and myeloid cells

- RAG-hu mouse model for Dengue

- 63% of mice develop antibodies when infected with DEN2

- Secondary infection with severe hemorrhagic symptoms possible

- Can also be transmitted via mosquito in the lab


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- HIV-1 infection of RAG-hu mice, long term viremia and gradual T cell decline (ip injection)

- Immune augmentation by PD-1 blockade? PD-1 is highly up-regulated on CD4 and CD8 T cells

- Anti-PD1 ligand (BMS)  2log reduction in viral load after 4 weeks of treatment

- Mucosal infection (drop of virus to vaginal or rectal wall)

- Preventive approach: Oral PrEP with raltegravir and maraviroc

-  Oral application for 3 days, mucosal challenge on day 4

- Maraviroc as a topical microbicide as a gel before sex, in mice applied 1 hour before challenge, mice were protected

- Application as a vaginal ring near cervix

- Antiviral action of anti-gp120 aptamer/siRNA chimera

- Binds to gp120 on infected cells and siRNA enters the cells

- After some time resistance to aptamer

- Chimera-treated mice eliminate viral load first, rebound after 7 weeks

- Take this technology to introduce a deadly siRNA and kill infected cells

Questions:

Which organ is target by Dengue? Liver cell is not infected but destroyed due to cytokine storm.

Can sperm interfere with the microbicide? Not tested.

X4 and R5 transmission the same? No comment.

Others could not see mucosal transmission? You have to wait for cells to accumulate in mucosa.

Beyond SARS: News from virus discovery and animal reservoir studies

Christian Drosten, University of Bonn Medical Centre

Lesions learned from Influenza field>Zoonosis: here, birds are key reservoir

Are Coronavirus related to Arthropod-Viruses?

So transmission: Arthropod>bat>human?

Two taxonomies:

Zoonotic “Sources” vs. evolutionary “origins” –ecological view

Connection to phylogenetic entry point if the virus is found on all branches of a monophyletic taxon

But very hard to prove that viruses derive from bat. You can say they pass to mammals a lot, but no proof of general origin.

Bats form huge colonies>good for viruses

- Virus/host adaptation is a plausible concept

- But are there any data suggesting bats deal with viruses in special ways?


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- We need data that go beyond phylogenies, in order to define /confirm reservoirs.

Reproductive success is one of the only readouts for pathogenicity of viruses in bats. Protected species.

Prediction

- How pathogenic are reservoir-borne Coronavirus?

- Do they still have to adapt before they can infect humans?

- Shouldn´t we screen pathogenicity factors carried by reservoir-borne viruses in high throughput assays?

Conclusions

- Small mammals host a large diversity of RNA viruses

- Virus detection is insufficient for risk assessment. We need to assess functional-, rather than genetic diversity

- Prediction might be an over-promise

- Immediate translational benefits could include animal surrogate models.

Emerging Bunyaviruses: SFTS

Masayuki Saijo, National Institute of Infectious Diseases-Tokyo

- Severe fever with thrombocytopenia syndrome in Japan

- Mysterious disease in mountainous regions Hubei and Henan 2008-9

- Fever, gastrointestinal symptoms, leukopenia and thrombocytopenia, and TBC

- Case fatality rate 21/171 12%

- Novel Bunyavirus SFTSV (genus Phlebovirus) causative agent, tick-borne Haemayphysalis longicornis

- 1 st patient, f, 50, no oversea travel, fever, general fatigue, vomiting, melena (bloody diarrhea)

- Leukocytes 400, thrombocytes 89.000/mm 3 in TBC, elevated AST, ALT. LDH, CK, ferritin suggestion for bacterial infection or leukemia  blood sample, virus isolation and PCR identified

+ EM

- Data collection from different centers, season April - December, western part of Japan, mostly

50-80 years old

A remembrance of Dr. Carlo Urbani

Guido Poli, San Raffaele University

Born: Castelplanio, Italy 1910-1956

Died: Bangkok (Thailand) 2-03-2003

WHO specialist in IDs in Hanoi, first examined patient for a suspected avian flu infection. Turned out to be SARS. Urbani developed SARS himself and died one month later.


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Saturday, June 1, 2013

Global Virus Network Semi-Annual Meeting

Seminar Room, Institute of Virology, Technische Universität München, Trogerstr. 30, 81675 Munich

9:00-10:00 Panel Discussion Part 1: Elimination of Hepatitis Viruses

Moderator: Ulrike Protzer, Munich

Franco Buonaguro, Italian Natl Cancer Institute

Worldwide disease burden of hepatitis viruses

HBV: Vaccination start 1991

Main HCV prevalence in Egypt and Tibet

Also Italy and particularly Sicily.

Treatment: Peg IFN and ribavirin,DAAs: Protease and polymerase inhibitors, miRNA inhibitors

Risk factors of HCC: HBV, HCV, TP53- and CTNNB1 mutations

Ulrike Protzer: Can cure of HCV infection be achieved?

Success story of science

89: detection; 99 replicon system; 05 cell culture system; 11 first DAA; 13 Cure?

Protease- and polymerase- and NS5A inhibitors

High cure rates can be achieved without IFN

HBV

5% of the world population infected, 25% of those die from HBV-related causes

Vertical transmission frequently leads to chronic infection;

This is a big epidemiologic problem, vertical transmission is difficult to prevent.

Ramou Njie, PROLIFICA: Chronic HBV infection in resource-limited settings

Clinical cases: Liver failure and HCC as important consequences for chronic HBV infection

Problems: No Africa guidelines for viral hepatitis

GHIS: Gambia Hepatitis Intervention Study

Phase 1: HBV Vaccination 2: HCC registry, HBV vaccine and HCC incidence 3: HCC surveillance

PROLIFICA: Prevention of liver cirrhosis and cancer in Africa

Matti Sällberg, Karolinska Institute, Sweden:

Elimination of hepatitis viruses by therapeutic vaccination

Worldwide prevalences of HBV and HCV

Therapeutic vaccine

HBV: viral vectors, HBsAg

HCV: viral vectors: Adenovirus, MVA


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Prime boost strategies for HCV

Conclusions: Therapeutic vaccination complements standard of care

Possible cure

Discussion part 1:

HBV: Therapeutic vaccine could save costs of antiviral treatment

HCV: no complicated algorithm for treatment initiation: Everybody should qualify for therapy

Recommended Focus of GVN: better diagnostic tests, studies to assess treatment effectiveness

HCV vaccines; HBV therapy and monitoring in resource-poor countries

10:30-12:30 Panel 2: Virus Outbreak Control (SARS, SFTS, H7N9, pathogen detection)

Moderators: Stefan Becker, Marburg, Germany & Billy Hall, CRID-UCD, Dublin Ireland.

Peter Palese, Mt. Sinai-NY USA; Christian Drosten, Uni Bonn, Germany; Hideki

Hasegawa, NIID-Tokyo, Japan; Yiming Shao, CDC-Beijing, China; Heinz Ellerbrok,

Robert Koch Inst-Berlin, Nicole Fischer, Hamburg, Gerd Sutter, LMU-Munich, Germany

Stefan Becker, Institute of Virology, Marburg

TTU emerging infections

Goals: Diagnostic preparedness; emergency vaccines; Detection of unknown pathogens

Christian Drosten, Institute of Virology, Bonn: New coronavirus

Basics on test validation to standards that industry accepts

Only structural proteins should be used for serology

IgM cross reactivity

Gerd Sutter, Institute of Veterinary Virology, Munich: MVA, a platform for development of emerging viruses

Vector developed and optimized over years

Production process under the Medical Devices Act

Recombinant H5 MVA including in clinical trial

Also suitable for H7N9 or MERS-CoV

Nicole Fischer, Hamburg-UKE: NGS for detection of pathogens

Worked well in retrospective analysis of extended gastroenteritis outbreak in East Germany

Quantification with PCR correlated with the number of recovered quasi-species

Cluster of unknown pneumonia

ARDS symptom in immunocompetent adults

RNA and DNA was extracted and respective libraries were prepared

Clamydophilia psitacci was detected in 2 patients, confirmed serologically


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Powerful tool to react to outbreaks

Important: validation of the tool

Needed: Annotated databases; databases for unknown sequences

Peter Palese, Mt. Sinai - NY USA: H7N9 influenza viruses

Human cases reported from China

Mostly elderly people affected

Summary of the present situation: No human transmission; no pandemic

Transmission to humans has probably happened in the past.

Heinz Ellerbrok, RKI, Berlin: Outbreak handling

Outbreak→Sample→Lab diagnostic: single/multiplex PCR, sequencing, electron microscopy

Exemplified time course of SARS epidemic

Mobile diagnostic devices, e.g. for NGS

Hideki Hasegawa, Tokyo, Japan: First autopsy findings of SFTS patients

Detection of SFTS RNA by in situ hybridization

Findings: Lymphadenitis, hypocellular bone marrow, SFTSV antigen detectable in tissue

IL6 secretion and apoptosis

Discussion

Systematic grant application; Money/grant distribution

Request from Nigeria: How to respond to Nigerians returning from Saudi Arabia with respiratory symptoms. (Dr. Abdulsalami Nasidi, Nigerian CDC)

Ulla Protzer, TU Munich: Summary of the panel discussion

Hepatitis panel:



prophylactic HCV vaccine;



broad HCV screening;



improve drug access in resource-limited settings;



goal: cure viral hepatitis;



Recommends GVN workshop to set up African hepatitis guidelines

Emerging viruses:



GVN well-prepared to react;



diagnostics and vaccine development;



expand sequence databases;



set up diagnostic tests for emerging viruses in Nigeria


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GVN Moscow Fall Meeting Update; Alexey Mazus, Moscow Center for HIV/AIDS Treatment &

Prevention. Dr. Mazus welcomed the GVN to Moscow. He provided a brief overview of GVN

Russia, noting the 8 Centers comprising GVN Russia.

Closing comments: GVN Scientific Director Gallo recognizes the many contributions of those who made the program a success. He noted in particular how many Board members participated at the entire meeting, showing their support for the GVN and their clear interest in understanding to the greatest degree possible the science behind the GVN. In her closing comments, GVN President Hrynkow urged scientists and Center Directors to be in good communication with her and other staff members. She will engage with donors on the specific projects and programs that need funding, but she needs to hear about the needs, gaps and opportunitie s to be seized in order to make timely and effective interventions. “Where is the pent up demand?”, she asked. She joined Dr. Gallo in thanking again Dr. Protzer and her team, all of the sponsors who made the meeting possible.


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GVNMunichNotes - Global Virus Network

Global Virus Network – Spring 2013 Meeting in Munich

Meeting Notes*

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Meeting Notes*

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