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Pseudotyped Viruses for Influenza

Del Rosario, Joanne Marie M. and da Costa, Kelly A.S. and Temperton, Nigel J. (2023) Pseudotyped Viruses for Influenza. In: Pseudotyped Viruses. Advances in Experimental Medicine and Biology . Springer Nature, pp. 153-173. ISBN 978-981-9901-12-8. (doi:10.1007/978-981-99-0113-5_8) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:100492)

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We have developed an influenza hemagglutinin (HA) pseudotype (PV) library encompassing all influenza A (IAV) subtypes from HA1-HA18, influenza B (IBV) subtypes (both lineages), representative influenza C (ICV), and influenza D (IDV) viruses. These influenza HA (or hemagglutinin-esterase fusion, HEF for influenza C/D) pseudotypes have been used in a pseudotype microneutralization assay, an optimized luciferase reporter assay, that is highly sensitive and specific for detecting neutralizing antibodies against influenza viruses. This has been an invaluable tool in detecting the humoral immune response against specific hemagglutinin or hemagglutinin-esterase fusion proteins for IAV to IDV in serum samples and for screening antibodies for their neutralizing abilities. Additionally, we have also produced influenza neuraminidase (NA) pseudotypes for IAV N1-N9 subtypes and IBV lineages. We have utilized these NA-PV as surrogate antigens in in vitro assays to assess vaccine immunogenicity. These NA-PV have been employed as the source of neuraminidase enzyme activity in a pseudotype enzyme-linked lectin assay (pELLA) that is able to measure neuraminidase inhibition (NI) titers of reference antisera, monoclonal antibodies, and postvaccination sera. Here we show the production of influenza HA, HEF, and NA-PV and their employment as substitutes for wild-type viruses in influenza serological and neutralization assays. We also introduce AutoPlate, an easily accessible web app that can analyze data from pMN and pELLA quickly and efficiently, plotting inhibition curves and calculating half-maximal concentration (IC50) neutralizing antibody titers. These serological techniques coupled with user-friendly analysis tools are faster, safer, inexpensive alternatives to classical influenza assays while also offering the reliability and reproducibility needed to advance influenza research and make it more accessible to laboratories around the world.

Item Type: Book section
DOI/Identification number: 10.1007/978-981-99-0113-5_8
Uncontrolled keywords: Influenza, Hemagglutinin, Neuraminidase, Pseudotypes, Neutralization, ELLA, Influenza vaccine, Influenza immunity, AutoPlate
Subjects: Q Science > QR Microbiology > QR355 Virology
Divisions: Divisions > Division of Natural Sciences > Medway School of Pharmacy
Funders: Bill & Melinda Gates Foundation (
Innovate UK (
Depositing User: Nigel Temperton
Date Deposited: 16 Mar 2023 00:31 UTC
Last Modified: 29 Mar 2023 13:31 UTC
Resource URI: (The current URI for this page, for reference purposes)

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