A comprehensive influenza reporter virus panel for high-throughput deep profiling of neutralizing antibodies

Adrian Creanga, Rebecca A. Gillespie, Brian E. Fisher, Sarah F. Andrews, Julia Lederhofer, Christina Yap, Liam Hatch, Tyler Stephens, Yaroslav Tsybovsky, Michelle C. Crank, Julie E. Ledgerwood, Adrian B. McDermott, John R. Mascola, Barney S. Graham () and Masaru Kanekiyo ()
Additional contact information
Adrian Creanga: National Institutes of Health
Rebecca A. Gillespie: National Institutes of Health
Brian E. Fisher: National Institutes of Health
Sarah F. Andrews: National Institutes of Health
Julia Lederhofer: National Institutes of Health
Christina Yap: National Institutes of Health
Liam Hatch: National Institutes of Health
Tyler Stephens: Frederick National Laboratory for Cancer Research sponsored by the National Cancer Institute
Yaroslav Tsybovsky: Frederick National Laboratory for Cancer Research sponsored by the National Cancer Institute
Michelle C. Crank: National Institutes of Health
Julie E. Ledgerwood: National Institutes of Health
Adrian B. McDermott: National Institutes of Health
John R. Mascola: National Institutes of Health
Barney S. Graham: National Institutes of Health
Masaru Kanekiyo: National Institutes of Health

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: Abstract Broadly neutralizing antibodies (bnAbs) have been developed as potential countermeasures for seasonal and pandemic influenza. Deep characterization of these bnAbs and polyclonal sera provides pivotal understanding for influenza immunity and informs effective vaccine design. However, conventional virus neutralization assays require high-containment laboratories and are difficult to standardize and roboticize. Here, we build a panel of engineered influenza viruses carrying a reporter gene to replace an essential viral gene, and develop an assay using the panel for in-depth profiling of neutralizing antibodies. Replication of these viruses is restricted to cells expressing the missing viral gene, allowing it to be manipulated in a biosafety level 2 environment. We generate the neutralization profile of 24 bnAbs using a 55-virus panel encompassing the near-complete diversity of human H1N1 and H3N2, as well as pandemic subtype viruses. Our system offers in-depth profiling of influenza immunity, including the antibodies against the hemagglutinin stem, a major target of universal influenza vaccines.

Date: 2021
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DOI: 10.1038/s41467-021-21954-2

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