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Pseudotyped Viruses for Influenza

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Pseudotyped Viruses

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1407))

Abstract

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 ICV and IDV) pseudotypes have been used in a pseudotype microneutralization assay (pMN), 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 to advance influenza research and make it more accessible to laboratories around the world.

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Authors and Affiliations

  1. Viral Pseudotype Unit, Medway School of Pharmacy, University of Kent and Greenwich at Medway, Chatham, UK

    Joanne Marie M. Del Rosario, Kelly A. S. da Costa & Nigel J. Temperton

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  1. Joanne Marie M. Del Rosario
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  2. Kelly A. S. da Costa
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  3. Nigel J. Temperton
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Correspondence to Nigel J. Temperton .

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  1. Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China

    Youchun Wang

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Del Rosario, J.M.M., da Costa, K.A.S., Temperton, N.J. (2023). Pseudotyped Viruses for Influenza. In: Wang, Y. (eds) Pseudotyped Viruses. Advances in Experimental Medicine and Biology, vol 1407. Springer, Singapore. https://doi.org/10.1007/978-981-99-0113-5_8

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