Cell mediated immunity against RSV and influenza in a human experimental challenge model

Lead Research Organisation: Imperial College London
Department Name: National Heart and Lung Institute

Abstract

Influenza and respiratory syncytial virus (RSV) are among the most common causes of lung infection. They affect around 664 million people each year worldwide, causing an estimated 500,000 and 160,000 deaths respectively. Current influenza vaccines need to change every year to overcome rapid viral mutation, and there is still no RSV vaccine. Further research is therefore urgently needed. T cells are important in controlling viral infections, but there is not enough known about their role in humans with influenza and RSV. Furthermore, there is evidence that the T cell response to RSV is not fully protective, leading to multiple infections with the same strain of RSV in some people.

This work will be carried out at the Centre for Respiratory Infection, Imperial College London within Professor Peter Openshaw‘s department. Healthy volunteers will be infected with either RSV or a weakened strain of influenza in order to examine the response of the T cells in their blood and lung secretions. By observing the changes in T cells following each infection, and comparing the responses to influenza and RSV, we aim to identify the essential components of an effective T cell response so that they can be used to create better vaccines.

Technical Summary

Background
Influenza and respiratory syncytial virus (RSV) are among the most common causes of viral respiratory infection, causing widespread morbidity and mortality particularly at the extremes of age and exacerbating conditions such as asthma. Vaccine development remains an urgent priority. While cell mediated immunity (CMI) is important in viral clearance and may enhance vaccine efficacy, its relative contribution in humans has not been fully determined. We aim to elucidate its role by comparing the human T cell responses to influenza and RSV. These differ in that re-infection with antigenically identical strains of influenza does not occur, while within 2 months of RSV challenge even subjects with high antibody levels may be re-infected with the same strain. The reasons for this are unclear but may involve immune modulation by viral products or impaired antigen presentation by RSV-infected dendritic cells leading to impaired T cell numbers function.

Hypotheses & aims
1. To test the hypothesis that RSV and influenza lead to quantitative and functional differences in T-cell responses.
2. To identify the mechanisms underlying poor antigen-specific CMI to RSV by comparing the transcriptional changes in T-cells following experimental challenge with RSV and live attenuated influenza vaccine (LAIV).

Study design
Healthy adult volunteers will be challenged with RSV Memphis 37 or LAIV. Samples will be collected periodically from blood, broncho-alveolar lavage (BAL) and nose. After 6 months, a second dose of virus will be administered and further samples obtained to examine the recall response.

The number of virus-specific T-cells will be analysed by ELISPOT and by MHC/peptide multimer staining. Differential effector functions will be studied including surface protein expression, cytokine production and proliferative capacity by CFSE staining. Flow cytometry will be used to characterize activation markers, co-stimulatory molecules, cytokines, and cytotoxic molecules. Nasal and bronchial fluid will be analysed for cytokine levels using multiplex bead arrays and Luminex. Using microarrays, we will identify transcriptional changes that occur during T-cell activation and following re-stimulation with RSV and LAIV. These data will recapitulate the predicted functional defects in RSV-infected T cells and will allow us to make inferences regarding underlying mechanisms via differential expression of signalling intermediates in TCR signal transduction.

CMI is important against both influenza and RSV infection, but protection against RSV is incomplete. By identifying correlates of immunity absent from the CMI response to RSV we aim to elucidate the basis for previous vaccine failures as well as novel approaches to vaccination.

Publications

10 25 50
 
Description WHO Working Group on RSV Vaccine Standardization and Clinical Evaluation
Geographic Reach Multiple continents/international 
Policy Influence Type Membership of a guideline committee
 
Description DARPA BAA Prometheus
Amount $800,000 (USD)
Organisation Defense Advanced Research Projects Agency (DARPA) 
Sector Public
Country United States
Start 03/2017 
End 08/2018
 
Description EU IMI Biovacsafe
Amount £17,425,663 (GBP)
Funding ID 115308 
Organisation European Commission 
Department Innovative Medicines Initiative (IMI)
Sector Public
Country Belgium
Start 03/2012 
End 03/2018
 
Description MRC GSK EMINENT
Amount £8,000,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 03/2017 
End 08/2019
 
Description Pilot research projects in human immunology
Amount $50,000 (USD)
Organisation National Institutes of Health (NIH) 
Sector Public
Country United States
Start 06/2011 
End 06/2012
 
Description Translation Award
Amount £2,772,791 (GBP)
Organisation Wellcome Trust 
Department Wellcome Trust Translation Award
Sector Charity/Non Profit
Country United Kingdom
Start 01/2016 
End 08/2018
 
Title RSV tetramers 
Description Only a few T cell targets in respiratory syncytial virus have been published. We defined a set of previously unrecognised CD4+ and CD8+ T cell targets and have made tetramers to label these cells for further analysis and isolation. 
Type Of Material Technology assay or reagent 
Year Produced 2015 
Provided To Others? Yes  
Impact Knowledge of these targets may enhance RSV vaccine development. 
 
Description Adaptive immune responses to viral vaccines and challenge 
Organisation Emory University
Department Emory Vaccine Centre
Country United States 
Sector Academic/University 
PI Contribution Study design, subject recruitment, sample collection, and carrying out assays.
Collaborator Contribution This collaboration has provided patient samples, sharing of techniques and strategic input.
Impact None
Start Year 2010
 
Description Full genome sequencing of RSV isolates 
Organisation Public Health England
Department Respiratory Virus Unit (RVU)
Country United Kingdom 
Sector Public 
PI Contribution Provision of respiratory samples from hospitalised patients and experimentally challenged adults for RSV full genome sequencing
Collaborator Contribution Full genome sequencing of RSV isolates
Impact None so far
Start Year 2014
 
Description Humoral responses to recombinant RSV proteins in experimental human infection 
Organisation Mucosis BV
Country Netherlands 
Sector Private 
PI Contribution Analysis of antibody and B cell responses to RSV F protein in adults challenged with RSV
Collaborator Contribution Provision of recombinant F proteins
Impact Paper submitted detailing the role of mucosal IgA in protection against RSV
Start Year 2012
 
Description Humoral responses to recombinant RSV proteins in experimental human infection 
Organisation Utrecht University
Department Department of Infectious Diseases and Immunology
Country Netherlands 
Sector Academic/University 
PI Contribution Analysis of antibody and B cell responses to RSV F protein in adults challenged with RSV
Collaborator Contribution Provision of recombinant F proteins
Impact Paper submitted detailing the role of mucosal IgA in protection against RSV
Start Year 2012
 
Description PREPARE WP3 
Organisation Amsterdam Medical Center
Department Department of Medical Microbiology and Infection Control
Country Netherlands 
Sector Hospitals 
PI Contribution Conduct of experimental human influenza infection study, collection of clinical samples, assessment of infection rate and clinical severity. Analysis of antibody and T cell responses.
Collaborator Contribution Transcriptomic analysis of blood and respiratory epithelial cells. Analysis of volatile organic compounds in exhaled breath.
Impact None as yet
Start Year 2015
 
Description Phase I and 2a clinical trial of Mucosis SynGEM 
Organisation Mucosis BV
Country Netherlands 
Sector Private 
PI Contribution Conducting phase 1 and phase 2a RSV challenge study clinical trials of a novel RSV vaccine SynGEM. Collection of clinical samples. Assessment of infection rate and severity. Analysis of antibody, B cell and T cell responses.
Collaborator Contribution Provision of SynGEM vaccine. Contracting of CRO and sponsorship of clinical trials. Analysis of serum samples by competition ELISA assays.
Impact None as yet
Start Year 2015
 
Description RNAseq of nasal curettage in experimental human RSV infection 
Organisation Merck
Country Germany 
Sector Private 
PI Contribution Collection of sequential nasal curettage samples from volunteers experimentally infected with RSV and assessment of infection status and clinical severity. Collaboration on analysis.
Collaborator Contribution RNA extraction and RNA sequencing of nasal curettage samples for transcriptomic analysis. Collaboration on analysis.
Impact None so far
Start Year 2015
 
Description Role of T follicular helper cells for cross-reactive antibody responses 
Organisation La Jolla Institute for Allergy & Immunology (LIAI)
Department Division of Translational Immunology
Country United States 
Sector Academic/University 
PI Contribution Study design, sample collection and performing assays.
Collaborator Contribution This collaboration has provided epitope predictions and techniques for the generation and use of peptide libraries.
Impact None
Start Year 2011
 
Description Role of cathelicidin in experimental human RSV infection 
Organisation Medical Research Council (MRC)
Department MRC Human Genetics Unit
Country United Kingdom 
Sector Academic/University 
PI Contribution Using the experimental human RSV model, we have collected serum and respiratory samples prior to and during infection with RSV, quantified viral shedding and determined the dynamics of disease.
Collaborator Contribution Cathelicidin levels have been measured in serum and bronchoalveolar lavage fluid. These will be correlated with clinical outcome and bronchial epithelial cell gene expression data.
Impact None so far.
Start Year 2013
 
Description Transcriptional and proteomic changes in human experimental RSV infection 
Organisation Duke University
Department Insitute of Genome Sciences and Policy
Country United States 
Sector Academic/University 
PI Contribution We have conducted experimental human infection of volunteers with RSV, collected blood and respiratory specimens for further transcriptomic and proteomic analysis.
Collaborator Contribution Our partners have provided GMP-certified RSV virus and will process and analyse blood and nasal samples for transcriptomic and proteomic analysis.
Impact None as yet
Start Year 2013
 
Description Whole blood transcriptomic signatures in experimental human infection with RSV 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Academic/University 
PI Contribution Blood samples provided for microarray analysis
Collaborator Contribution Processing of RNA for microarray
Impact None so far
Start Year 2012
 
Description Imperial Festival 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Attended stall publicising our clinical research work with large models showing the nose and respiratory tract mucosa as well as games and multimedia displays focusing on influenza and the MOSAIC project. The Imperial Festival lasts an entire weekend and 344 e-surveys were collated. The majority of respondees felt that the event had developed their interest in science "a lot" and 88% would recommend the Festival to others.
Year(s) Of Engagement Activity 2013,2014,2015