A Comprehensive Influenza Reporter Virus Panel for High-throughput Deep Profiling of Neutralizing Antibodies

Overview

Journal Nat Commun

Specialty Biology

Date 2021 Mar 20

PMID 33741916

Citations 32

Authors

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

Masaru Kanekiyo

Affiliations

Soon will be listed here.

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.

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