Exploiting Pan Influenza A and Pan Influenza B Pseudotype Libraries for Efficient Vaccine Antigen Selection
Joanne Marie M. Del Rosario,
Kelly A. S. da Costa,
Benedikt Asbach,
Francesca Ferrara,
Matteo Ferrari,
David A. Wells,
Gurdip Singh Mann,
Veronica O. Ameh,
Claude T. Sabeta,
Ashley C. Banyard,
Rebecca Kinsley,
Simon D. Scott,
Ralf Wagner,
Jonathan L. Heeney,
George W. Carnell,
Nigel J. Temperton
Affiliations
Joanne Marie M. Del Rosario
Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway, Chatham ME4 4BF, UK
Kelly A. S. da Costa
Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway, Chatham ME4 4BF, UK
Benedikt Asbach
Institute of Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany
Francesca Ferrara
Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway, Chatham ME4 4BF, UK
Matteo Ferrari
DIOSynVax, Cambridge CB3 0ES, UK
David A. Wells
DIOSynVax, Cambridge CB3 0ES, UK
Gurdip Singh Mann
Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway, Chatham ME4 4BF, UK
Veronica O. Ameh
Department of Veterinary Public Health and Preventive Medicine, College of Veterinary Medicine, Federal University of Agriculture Makurdi, Makurdi P.M.B. 2373, Bene State, Nigeria
Claude T. Sabeta
Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, P. Bag X04, Onderstepoort 0110, South Africa
Ashley C. Banyard
Animal and Plant Health Agency (APHA), Department of Virology, Weybridge, Surrey KT15 3NB, UK
Rebecca Kinsley
DIOSynVax, Cambridge CB3 0ES, UK
Simon D. Scott
Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway, Chatham ME4 4BF, UK
Ralf Wagner
Institute of Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany
Jonathan L. Heeney
DIOSynVax, Cambridge CB3 0ES, UK
George W. Carnell
DIOSynVax, Cambridge CB3 0ES, UK
Nigel J. Temperton
Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway, Chatham ME4 4BF, UK
We developed an influenza hemagglutinin (HA) pseudotype library encompassing Influenza A subtypes HA1-18 and Influenza B subtypes (both lineages) to be employed in influenza pseudotype microneutralization (pMN) assays. The pMN is highly sensitive and specific for detecting virus-specific neutralizing antibodies against influenza viruses and can be used to assess antibody functionality in vitro. Here we show the production of these viral HA pseudotypes and their employment as substitutes for wildtype viruses in influenza neutralization assays. We demonstrate their utility in detecting serum responses to vaccination with the ability to evaluate cross-subtype neutralizing responses elicited by specific vaccinating antigens. Our findings may inform further preclinical studies involving immunization dosing regimens in mice and may help in the creation and selection of better antigens for vaccine design. These HA pseudotypes can be harnessed to meet strategic objectives that contribute to the strengthening of global influenza surveillance, expansion of seasonal influenza prevention and control policies, and strengthening pandemic preparedness and response.