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J Gen Virol. 2014 Jul 7. pii: vir.0.067413-0. doi: 10.1099/vir.0.067413-0. [Epub ahead of print]
Exploring the antigenic relatedness of influenza virus hemagglutinins with strain-specific polyclonal antibodies.
Garc?a-Barreno B1, Delgado T1, Benito S1, Casas I2, Pozo F2, Melero JA3.
Author information
Abstract
Alternative methods to the standard hemagglutination inhibition (HI) and neutralization tests to probe the antigenic properties of the influenza virus hemagglutinin (HA) were developed in this study. Vaccinia virus recombinants expressing reference HAs were used to immunize rabbits from which polyclonal antibodies were obtained. These antibodies were subtype specific but showed limited intra-subtype strain specificity in ELISA. The discriminatory capacity of these antibodies was however markedly increased after adsorption to cells infected with heterologous influenza viruses, revealing antigenic differences that were otherwise undistinguishable by standard HI and neutralization tests. Furthermore, the unadsorbed antibodies could be used to select escape mutants of the reference strain which after sequencing unveiled amino acid changes responsible of the noted antigenic differences. These procedures therefore provide alternative methods for the antigenic characterization of influenza HA and might be useful in studies of HA antigenic evolution.
Copyright ? 2014, the Society for General Microbiology.
KEYWORDS:
antigenicity; hemagglutinin; infuenza
PMID:
25000959
[PubMed - as supplied by publisher]
Exploring the antigenic relatedness of influenza virus hemagglutinins with strain-specific polyclonal antibodies.
Garc?a-Barreno B1, Delgado T1, Benito S1, Casas I2, Pozo F2, Melero JA3.
Author information
Abstract
Alternative methods to the standard hemagglutination inhibition (HI) and neutralization tests to probe the antigenic properties of the influenza virus hemagglutinin (HA) were developed in this study. Vaccinia virus recombinants expressing reference HAs were used to immunize rabbits from which polyclonal antibodies were obtained. These antibodies were subtype specific but showed limited intra-subtype strain specificity in ELISA. The discriminatory capacity of these antibodies was however markedly increased after adsorption to cells infected with heterologous influenza viruses, revealing antigenic differences that were otherwise undistinguishable by standard HI and neutralization tests. Furthermore, the unadsorbed antibodies could be used to select escape mutants of the reference strain which after sequencing unveiled amino acid changes responsible of the noted antigenic differences. These procedures therefore provide alternative methods for the antigenic characterization of influenza HA and might be useful in studies of HA antigenic evolution.
Copyright ? 2014, the Society for General Microbiology.
KEYWORDS:
antigenicity; hemagglutinin; infuenza
PMID:
25000959
[PubMed - as supplied by publisher]
