Influenza A (N1-N9) and Influenza B (B/Victoria and B/Yamagata) Neuraminidase Pseudotypes as Tools for Pandemic Preparedness and Improved Influenza Vaccine Design
Kelly A. S. da Costa,
Joanne Marie M. Del Rosario,
Matteo Ferrari,
Sneha Vishwanath,
Benedikt Asbach,
Rebecca Kinsley,
Ralf Wagner,
Jonathan L. Heeney,
George W. Carnell,
Nigel J. Temperton
Affiliations
Kelly A. S. da Costa
Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway, Chatham ME4 4BF, UK
Joanne Marie M. Del Rosario
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
Sneha Vishwanath
Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK
Benedikt Asbach
Institute of Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany
Rebecca Kinsley
DIOSynVax, Cambridge CB3 0ES, 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
Department of Veterinary Medicine, University of Cambridge, 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
To better understand how inhibition of the influenza neuraminidase (NA) protein contributes to protection against influenza, we produced lentiviral vectors pseudotyped with an avian H11 hemagglutinin (HA) and the NA of all influenza A (N1–N9) subtypes and influenza B (B/Victoria and B/Yamagata). These NA viral pseudotypes (PV) possess stable NA activity and can be utilized as target antigens in in vitro assays to assess vaccine immunogenicity. Employing these NA PV, we developed an enzyme-linked lectin assay (pELLA) for routine serology to measure neuraminidase inhibition (NI) titers of reference antisera, monoclonal antibodies and post-vaccination sera with various influenza antigens. We also show that the pELLA is more sensitive than the commercially available NA-Fluor™ in detecting NA inhibition in these samples. Our studies may lead to establishing the protective NA titer that contributes to NA-based immunity. This will aid in the design of superior, longer lasting and more broadly protective vaccines that can be employed together with HA-targeted vaccines in a pre-pandemic approach.
