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J Vis Exp. 2017 Aug 29;(126). doi: 10.3791/56067.
Generation of Escape Variants of Neutralizing Influenza Virus Monoclonal Antibodies.
Leon PE1, Wohlbold TJ1, He W1, Bailey MJ1, Henry CJ2, Wilson PC2, Krammer F3, Tan GS4.
Author information
Abstract
Influenza viruses exhibit a remarkable ability to adapt and evade the host immune response. One way is through antigenic changes that occur on the surface glycoproteins of the virus. The generation of escape variants is a powerful method in elucidating how viruses escape immune detection and in identifying critical residues required for antibody binding. Here, we describe a protocol on how to generate influenza A virus escape variants by utilizing human or murine monoclonal antibodies (mAbs) directed against the viral hemagglutinin (HA). With the use of our technique, we previously characterized critical residues required for the binding of antibodies targeting either the head or stalk of the novel avian H7N9 HA. The protocol can be easily adapted for other virus systems. Analyses of escape variants are important for modeling antigenic drift, determining single nucleotide polymorphisms (SNPs) conferring resistance and virus fitness, and in the designing of vaccines and/or therapeutics.
PMID: 28872136 DOI: 10.3791/56067
Generation of Escape Variants of Neutralizing Influenza Virus Monoclonal Antibodies.
Leon PE1, Wohlbold TJ1, He W1, Bailey MJ1, Henry CJ2, Wilson PC2, Krammer F3, Tan GS4.
Author information
Abstract
Influenza viruses exhibit a remarkable ability to adapt and evade the host immune response. One way is through antigenic changes that occur on the surface glycoproteins of the virus. The generation of escape variants is a powerful method in elucidating how viruses escape immune detection and in identifying critical residues required for antibody binding. Here, we describe a protocol on how to generate influenza A virus escape variants by utilizing human or murine monoclonal antibodies (mAbs) directed against the viral hemagglutinin (HA). With the use of our technique, we previously characterized critical residues required for the binding of antibodies targeting either the head or stalk of the novel avian H7N9 HA. The protocol can be easily adapted for other virus systems. Analyses of escape variants are important for modeling antigenic drift, determining single nucleotide polymorphisms (SNPs) conferring resistance and virus fitness, and in the designing of vaccines and/or therapeutics.
PMID: 28872136 DOI: 10.3791/56067