Two Escape Mechanisms of Influenza A Virus to a Broadly Neutralizing Stalk-Binding Antibody

• DOI: 10.1371/journal.ppat.1005702
• Publication Date: 2016-06-28T17:29:48Z
• Authors: Ning Chai; Lee R. Swem; Mike Reichelt; Haiyin Chen-Harris; Elizabeth Luis; Summer Park; Ashley Fouts; Patrick Lupardus; Thomas D. Wu; Olga Li; Jacqueline McBride; Michael Lawrence; Min Xu; Man-Wah Tan
• Keywords: 0301 basic medicine; QH301-705.5; Blotting, Western; Hemagglutinin Glycoproteins, Influenza Virus; Antibodies, Viral; Polymerase Chain Reaction; Madin Darby Canine Kidney Cells; Mice; 03 medical and health sciences; Dogs; Orthomyxoviridae Infections; Neutralization Tests; Influenza, Human; Animals; Humans; Biology (General); Immune Evasion; Drug Resistance, Microbial; RC581-607; Flow Cytometry; Antibodies, Neutralizing; Immunohistochemistry; 3. Good health; Influenza A virus; Immunologic diseases. Allergy; Research Article

Broadly neutralizing antibodies targeting the stalk region of influenza A virus (IAV) hemagglutinin (HA) are effective in blocking virus infection both in vitro and in vivo. The highly conserved epitopes recognized by these antibodies are critical for the membrane fusion function of HA and therefore less likely to be permissive for virus mutational escape. Here we report three resistant viruses of the A/Perth/16/2009 strain that were selected in the presence of a broadly neutralizing stalk-binding antibody. The three resistant viruses harbor three different mutations in the HA stalk: (1) Gln387Lys; (2) Asp391Tyr; (3) Asp391Gly. The Gln387Lys mutation completely abolishes binding of the antibody to the HA stalk epitope. The other two mutations, Asp391Tyr and Asp391Gly, do not affect antibody binding at neutral pH and only slightly reduce binding at low pH. Interestingly, they enhance the fusion ability of the HA, representing a novel mechanism that allows productive membrane fusion even in the presence of antibody and hence virus escape from antibody neutralization. Therefore, these mutations illustrate two different resistance mechanisms used by IAV to escape broadly neutralizing stalk-binding antibodies. Compared to the wild type virus, the resistant viruses release fewer progeny viral particles during replication and are more sensitive to Tamiflu, suggesting reduced viral fitness.