08 September

Lady Brodie Room, Hall Building, St Hilda's College

9.15 - 09.45: Registration, coffee and networking

09.45: Welcome and housekeeping

09.50: Opening by symposium co-chairs

Session 1: Chair Dr Nigel Temperton

10.00: Professor Sarah Gilbert (Keynote Speaker)
University of Oxford, UK
Measurement of immune responses in the development of improved influenza vaccines

10.40: Dr Karen Laurie
WHO Collaborating Centre for Influenza, Peter Doherty Institute for Infection and Immunity, Melbourne, Australia
Improving standardisation and timeliness of Seroepidemiological studies through the Global Partnership CONSISE (the Consortium for the Standardization of Influenza Seroepidemiology)

11.00: Dr Qibo Zhang
University of Liverpool, UK
Cross-reactive immunity against influenza viruses in children and adults following 2009 pandemic H1N1 infection 

11.20: Dr Adrian Shepherd
Birkbeck College (University of London), UK
Immunoinformatics and computational virology: making a contribution to surveillance and vaccine design

11.50: Miss Francesca Ferrara
University of Kent, UK
Pseudotypes as tools to study heterosubtypic immunity

12.10: Dr I-Na Lu
National Health Laboratory, Luxembourg
Generation of hemagglutinin stalk specific CD4+ T cell immune responses during sublethal dose influenza A virus infection

12.30: Professor Xiao-Ning Xu (Keynote Speaker)
Chair in Human Immunology, Department of Medicine, Imperial College London, UK
T-cell responses in the human flu challenge model

13.10: Lunch

Session 2: Chair Dr Simon Scott

14.00: Dr Geraldine Taylor
Vaccinology group, The Pirbright Institute, UK
Replication-Defective Chimpanzee Adenovirus and MVA Vectors Expressing Human (H)RSV Antigens Protect Calves Against (B)RSV Infection

14.30: Miss Rebecca Kinsley
University of Kent, UK
The contribution of specific haemagglutinin mutations to equine influenza vaccine breakdown

14.40: Dr David Griffiths
Moredun Research Institute, Scotland, UK
Retrovirus-induced lung cancer in sheep

15.00: Professor Maria Zambon
Director, Reference Microbiology Laboratory, Public Health England, UK
Middle East respiratory syndrome coronavirus

15.40: Coffee, networking

16.10: Dr Kerstin Beer
MRC National Institute for Medical Research, UK
Generation and characterisation of monoclonal antibodies recognising neutralising epitopes of currently circulating Influenza A (H3N2) viruses

16.30: Dr Lynda Coughlan
University of Oxford, UK
An update from clinical trials using viral vectored vaccines for influenza

16.50: Dr Richard Pebody
Public Health England, London, UK
The seroepidemiology of acute respiratory viruses: what is the public health rationale?

17.20: Closing remarks

19.30:  Speakers dinner at St Aldates Tavern, Oxford (sponsored by Viral Pseudotype Unit)

Detailed Agenda

08 September

Lady Brodie Room, Hall Building, St Hilda's College

9.15 - 09.45: Registration, coffee and networking

09.45: Welcome and housekeeping

09.50: Opening by symposium co-chairs

Session 1: Chair Dr Nigel Temperton

10.00: Professor Sarah Gilbert (Keynote Speaker)
University of Oxford, UK
Measurement of immune responses in the development of improved influenza vaccines

Abstract: The methods for determining immune response to licensed influenza vaccines have changed very little over past decades. However with the development of novel vaccines designed to induce broader immunity, whether within clade or cross clade immunity, new assays are also being developed and implemented. The methods now in use for testing new influenza vaccines in humans, pigs and poultry will be discussed.

10.40: Dr Karen Laurie
WHO Collaborating Centre for Influenza, Peter Doherty Institute for Infection and Immunity, Melbourne, Australia
Improving standardisation and timeliness of Seroepidemiological studies through the Global Partnership CONSISE (the Consortium for the Standardization of Influenza Seroepidemiology

Abstract: CONSISE, the Consortium for the Standardization of Influenza Seroepidemiology, is a global partnership to develop investigation protocols and standardise seroepidemiology to inform health policy for influenza and other respiratory diseases. CONSISE is comprised of an epidemiology and laboratory working group. The laboratory working group is focused on serological assay standardisation and has developed consensus assay protocols and performed international laboratory comparisons of serological assays to detect antibodies to influenza viruses. Epidemiological and serological protocols developed by CONSISE members have recently been used ‘in the field’ to study MERS-CoV and A(H7N9) outbreaks. CONSISE’s materials are open access and we encourage membership from all interested parties. This applied research may be of interest to others at the Respiratory Viruses 2014 meeting performing such collaborative studies to improve seroepidemiology. 

11.00: Dr Qibo Zhang
University of Liverpool, UK
Cross-reactive immunity against influenza viruses in children and adults following 2009 pandemic H1N1 infection

Muhammad S Ahmed¹, Laura C Jacques¹, Waleed Mahallawi¹, Francesca Ferrara², Nigel Temperton^2, Nav Upile³, Casey Vaughan³, Ravi Sharma³, Helen Beer⁴, Katja Hoschler⁵, Paul S McNamara⁶, Qibo Zhang¹

Institute of Infection and Global Health, University of Liverpool¹, University of Kent^2, ENT Department, Alder Hey Children’s Hospital³; ENT Department, Royal Liverpool University Hospital⁴, Public Health England, London⁵,  Department of Women’s and Children’s Health, University of Liverpool⁶

Abstract: 2009 H1N1 pandemic influenza (A(H1N1)pdm09) resulted in considerable morbidity and mortality in humans. Recent studies suggest infection with A(H1N1)pdm09 virus elicited cross-reactive anti-hemagglutinin (HA) memory B cell response to conserved regions of HA. However, the breadth and magnitude of cross-reactive immunity in children and adults following A(H1N1)pdm09 infection are unknown. Methods: We investigated serum anti-HA immunity to a number of group-1 and -2 viruses in children and adults following 2009 pandemics using hemagglutination inhibition (HAI), enzyme-linked immunosorbent assay and virus neutralization assay. Results: Applying hemagglutination inhibition (HAI) titres≥40 against A(H1N1)pdm09 as threshold of sero-positivity, we observed significantly higher levels of anti-HA antibodies to a number of virus subtypes, including those neutralizing H5N1, in subjects with evidence of exposure to pandemic H1N1 than those without. Adults demonstrated broader and stronger cross-reactive anti-HA antibodies than children, including cross-reactive anti-HA1 and -HA2 antibodies. By comparison, individuals with serologic evidence of recent exposure to seasonal H1N1 or H3N2 did not show such broad cross-reactive immunity. Conclusion: Our results suggest individuals exposed to A(H1N1)pdm09 virus developed a broad and age-associated cross-reactive anti-HA immunity which may have important implications for future vaccination strategies to enable protection against a broader range of influenza viruses.

11.20: Dr Adrian Shepherd
Birkbeck College (University of London), UK
Immunoinformatics and computational virology: making a contribution to surveillance and vaccine design

Abstract: We analyze large sets of sequences, often mapped to structural data, to gain insights into the antigenic escape of viral populations driven by herd immune responses. For example, using our "epitope clustering" method, we aim to predict escape from prevalent antibodies that target influenza A, and have challenged assumptions about "conserved" vaccine targets on the stalk of haemagglutinin and the shielding role of N-glycans. We also analyse antibody repertoires from Next Generation Sequencing data, focusing on two rapidly mutating viruses, influenza A and HIV. The ultimate aim is to support the design of immunogens capable of guiding the host immune response down a particular maturation pathway that culminates in the induction of broadly neutralising antibodies.

11.50: Miss Francesca Ferrara
University of Kent, UK
Pseudotypes as tools to study heterosubtypic immunity

Abstract: Currently unavailable

12.10: Dr I-Na Lu
National Health Laboratory, Luxembourg
Generation of hemagglutinin stalk specific CD4+ T cell immune responses during sublethal dose influenza A virus infection 

I-Na Lu, Sophie Farinelle, Claude P Muller

Laboratory of Immunology, Centre de Recherche Public de la Santé (CRP-Santé), Luxembourg 

Abstract: The stalk region of the influenza virus hemagglutinin (HA) is relatively well conserved when comparing to the globular head domain of HA, and antisera elicited against the HA stalk region are shown to be cross-protective between strains, which makes it a potential target for the universal vaccine against influenza. However, despite these encouraging results, studies have not yet determined the role of CD4+ T cells, which might serve as a vital player in mediating HA stalk-specific humoral and cellular immune responses. To better understand the CD4+ T cell immunogenicity of the HA stalk, which might be important for protection from influenza virus infection, we sought to determine the profile of HA stalk-specific CD4+ T cell responses generated during sublethal infection of influenza. Using a model of murine influenza virus infection, significantly increased activation and differentiation of T follicular helper (Tfh) cells were observed together with the expansion of germinal center (GC) B cells in the lung draining lymph nodes (dLNs). However, the splenocytes from only half of the infected mice produced IL-2 and IFN-γ after HA stalk-specific CD4+ T cell stimulation measured by the enzyme-linked immunospot (ELISpot) assay. The induction of polyfunctional CD4+ T cell subsets was significantly lower in magnitude by using the HA stalk peptide pool when compared to using the HA recombinant protein. Overall, our data showed that HA stalk induced low magnitude of CD4+ T cell responses following sublethal infection of influenza. In order to improve cross-strain protection against influenza virus infections, the generation of broad and highly polyfunctional CD4+ T cell responses might be important for the future vaccine design.

12.30: Professor Xiao-Ning Xu (Keynote Speaker)
Chair in Human Immunology, Department of Medicine, Imperial College London, UK
T-cell responses in the human flu challenge model

Abstract: While neutralizing antibodies offer major protective immunity against influenza infection, the precise role of T cells remains unclear in humans. To address this issue, we conducted influenza human challenge studies and studied the role of pre-existing and developing T cell immunity in limiting illness severity. We infected healthy volunteers with no detectable antibodies to the challenge viruses (live seasonal H3N2 or H1N1 viruses). Our studies identify a subset of CD4⁺ T cells that is an important correlate of both homotypic and heterotypic protection and also provides potentially valuable information for the design of future vaccines against emerging strains.

13.10: Lunch

Session 2: Chair Dr Simon Scott

14.00: Dr Geraldine Taylor
Vaccinology group, The Pirbright Institute, UK
Replication-Defective Chimpanzee Adenovirus and MVA Vectors Expressing Human (H)RSV Antigens Protect Calves Against (B)RSV Infection

Geraldine Taylor¹, Michelle Thom¹, Becky Herbert¹, Efrain Guzman¹, Angiolo Pierantoni²,  Stefania Capone², Stefano Colloca², Riccardo Cortese³,  Alfredo Nicosia^2,4,5 and Alessandra Vitelli²

¹The Pirbright Institute, Ash Road, Pirbright, Surrey, GU24 0NF, UK; ²Okairòs, Naples, Italy; ³Keires AG, Basel, Switzerland; ⁴University of Naples Federico II, Italy; ⁵Ceinge - Biotecnologie Avanzate s.c.a r.l., Naples, Italy

Abstract: We have evaluated the safety, immunogenicity and efficacy of a novel vectored HRSV vaccine in calves, a model of natural RSV infection. Bovine (B)RSV is a major cause of respiratory disease in young calves and is genetically and antigenically closely related to HRSV. We have generated a novel genetic vaccine consisting of an artificial, consensus-based, HRSV antigen which includes a soluble F protein for the induction of neutralizing antibodies and the conserved N and M2-1 internal proteins, which are known to elicit T cell immunity in humans. This complex antigen is encoded by a replication incompetent chimpanzee Adenovirus (Ad), to which there is limited pre-existing immunity in man, and by Modified Vaccinia Ankara (MVA) vectors. The vaccine candidates, which induce protective immunity in mice and cotton rats, were administered to young, BRSV sero-negative calves in different combinations of homologous or heterologous prime/boost regimens, with the priming and boosting vector being administered either intramuscularly (IM) or intranasally (IN). Calves were challenged IN and intratracheally with BRSV, 4 weeks after vaccination.  Following challenge, nasopharyngeal excretion of BRSV was monitored daily and at study end, 6 days after BRSV challenge, BRSV titres in the lung and the extent of pulmonary pathology were evaluated. The vaccines were immunogenic and induced complete protection against nasal BRSV replication in calves when administered by Ad-RSV IN prime/MVA-RSV IM boost regimen. Complete protection against replication of BRSV in the lungs was also induced by Ad IN/Ad IM, Ad IM/Ad IM and Ad IM/MVA IM regimens. Complete protection against pulmonary pathology was induced by Ad IN/MVA IM, and the extent of pulmonary pathology was significantly reduced in calves vaccinated with all regimens. These studies demonstrate that complete, cross-protective efficacy was induced in calves by a novel virus-vectored HRSV vaccine and there was no evidence of vaccine-enhanced disease.

14.30: Miss Rebecca Kinsley
University of Kent, UK
The contribution of specific haemagglutinin mutations to equine influenza vaccine breakdown

Rebecca Kinsley¹, Stuart Mather¹, Debra Elton², Alana Kilby², Janet Daly ³, Nigel Temperton¹ & Simon Scott¹

¹Viral Pseudotype Unit, School of Pharmacy, University of Kent, Central Avenue, Chatham Maritime, ME4 4TB; ²Animal Health Trust, Lanwades Park, Kentford, Newmarket, Suffolk, CB8 7UU; ³School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington, LE12 5RD 

Abstract: In 1979, influenza caused a major epidemic amongst horses across Europe including Newmarket, UK. Subsequently, vaccines were produced using multiple outbreak strains including A/equine/Newmarket/79 and A/equine/Fontainebleau/1/79 (both subtype H3N8) which prevented further outbreaks until 1989 when a new antigenic drift variant emerged. The A/equine/Sussex/89 (H3N8) strain came from one of the affected regions in the UK where both unvaccinated and, notably, vaccinated horses were affected. The accumulation of mutations within important antigenic epitopes of the virus surface glycoprotein haemagglutinin (HA) can lead to a decrease in the efficiency of antibody recognition.  Three mutations, previously identified within five major epitopes of human influenza HA, have been incorporated into equine influenza pseudotyped lentiviruses (PVs). The PVs were generated via co-transfection of HEK293T cells with four plasmids expressing the equine influenza HA surface glycoproteins, HIV gag-pol, firefly luciferase reporter gene and TMPRSS2 endoprotease (to cleave the HA). The resulting viruses were harvested and pseudotype virus antibody neutralisation assays (PVNAs) against Newmarket/79 specific sera were performed.  Results showed that specific single amino acid mutations in the putative major epitope sites altered the ability of the sera to neutralise the PVs, indicating their importance in vaccine protection.

14.40: Dr David Griffiths
Moredun Research Institute, Scotland, UK
Retrovirus-induced lung cancer in sheep

David J Griffiths, Charline Alleaume, Esther Bijsmans, Henny M Martineau, Patricia Dewar, Jeanie Finlayson, Mark P Dagleish, Chris Cousens

Moredun Research Institute, Pentlands Science Park, Edinburgh, EH26 0PZ, UK

Abstract: Ovine pulmonary adenocarcinoma (OPA) is a common infectious respiratory disease of sheep caused by jaagsiekte sheep retrovirus (JSRV) and is an important economic and animal welfare problem for sheep farmers.  In addition, OPA is regarded as a valuable natural animal model for human lung adenocarcinoma.  Research in our laboratory is focused on improving understanding of the pathogenesis of OPA in order to inform the development of diagnostic tests that can be used to control the disease and reduce its impact on sheep farming.  Recent studies have identified the presence of neutralizing antibodies to JSRV in naturally affected sheep and identified candidate tumour antigens that have potential application as diagnostic markers.  We have also developed a novel in vitro lung tumour model that reproduces many of the features of JSRV infection of lung tissue observed in vivo.  This model provides a platform for future studies of early events in OPA oncogenesis and enhances the utility of OPA as a model for human lung adenocarcinoma.  

15.00: Professor Maria Zambon
Director, Reference Microbiology Laboratory, Public Health England, UK
Middle East respiratory syndrome coronavirus

Abstract: Currently unavailable

15.40: Coffee, networking

16.10: Dr Kerstin Beer
MRC National Institute for Medical Research, UK
Generation and characterisation of monoclonal antibodies recognising neutralising epitopes of currently circulating Influenza A (H3N2) viruses 

Kerstin Beer, Rodney S Daniels, John W McCauley

MRC National Institute for Medical Research, Division of Virology, London NW7 1AA, UK

Abstract: Influenza is an infectious disease of the respiratory tract which is preventable by vaccination. The main target of vaccine-induced neutralising antibodies (Abs) is the surface protein haemagglutinin (HA). Sequence comparisons show a number amino acid changes in the HA protein during the evolution of human influenza A (H3N2) viruses which prevents them from being recognized by neutralising Abs. A monoclonal Ab (mAb) recognising a neutralising epitope, which is conserved in a broad spectrum of influenza viruses, would have the potential to prevent and treat influenza, regardless of the causal strain. In this work a panel of 12 mouse mAbs recognising neutralising epitopes of current circulating H3N2 viruses was generated by using the hybridoma technology. A virus neutralising assay and a heamagglutination inhibition assay showed that almost all of the neutralising mAbs inhibit the receptor binding of recent H3N2 influenza viruses. Viruses isolated prior to 2006, a time after which a decrease in receptor avidity was seen, were not recognised. The neutralising epitopes have been identified by generating mAb escape viruses. Sequence analyses of HA of the escape viruses have determined the haemagglutinin head domain as the main target of the neutralising mAbs. Further structural analyses of the neutralising epitopes are planned. The generation and characterisation of mAbs neutralising a broad spectrum of current circulating H3N2 influenza virus isolates provides important information for the development of immunotherapy of influenza infections. Information about identified neutralising epitopes which are conserved in a broad spectrum of currently circulating H3N2 viruses can be used to design vaccines which induce the production of broadly neutralising antibodies against influenza viruses. 

16.30: Dr Lynda Coughlan
Postdoctoral Researcher, Jenner Institute, University of Oxford, UK
An update from clinical trials using viral vectored vaccines for influenza

Coughlan L, Payne R, Milicic A, Venkatraman N, Antrobus R, Lugonja B, Hill AV, Lambe T, Gilbert SC

Abstract: Human Adenovirus Type-5 (Ad5) vectors are used most commonly in clinical trials. Adenoviruses (Ads) are promising vaccine vectors for infectious diseases due to their ability to induce and boost cellular immunity to transgene antigens. However, high level pre-existing immunity in humans has the potential to limit the efficacy of Ad5-based vaccines. In recent years this issue has prompted the investigation of rare species or non-human Ad vectors. We report the first dose-escalation Phase I clinical trial to determine safety and immunogenicity of replication-defective chimpanzee Ad, ChAdOx1, expressing conserved antigens from Influenza A, nucleoprotein (NP) and matrix protein-1 (M1). There were no serious adverse events at any dose and the vaccine was well tolerated up to 2.5x10¹⁰vp. At the highest dose (5x10¹⁰vp), 3/6 volunteers experienced adverse reactions including fever, fatigue, malaise and headache. Peak T-cell responses (ELISpot) to NP+M1 peptide pools were observed D14 post-vaccination and were increased ~3-fold (5x10⁸vp), 6-fold (5x10⁹vp), 7-fold (2.5x10¹⁰vp) and 4-fold (5x10¹⁰vp) over baseline (D0). A small number of volunteers received a boost vaccination with poxviral vector MVA-NP+M1 ~8 weeks post-prime with ChAdOx1-NP+M1. MVA-NP+M1 boosted T-cell responses to levels comparable to the initial ChAdOx1-NP+M1 prime vaccination. In a separate clinical trial we are currently assessing the comparative immunogenicity of ChAdOx1-NP+M1 and MVA-NP+M1 in a prime-boost regimen, which includes four groups receiving boost vaccinations at an interval of 8 or 52 weeks post initial vaccination. Ongoing data from this trial will be presented.

Keywords: adenovirus, vaccine, influenza, clinical

16.50: Dr Richard Pebody
Public Health England, London, UK
The seroepidemiology of acute respiratory viruses: what is the public health rationale?

Abstract: Using examples from established infections such as seasonal influenza plus new and emerging pathogens such as pandemic influenza and mers -cov, this presentation will outline how seroepidemiology can be used to address important public health questions. 

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