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Production and characterization of monoclonal antibodies against the hemagglutinin of H5N1 and antigenic investigation of avian influenza H5N1 viruses isolated from China

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Abstract:

Eight monoclonal antibodies against hemagglutinin of influenza A virus A/Chicken/Henan/01/2004(H5N1) were produced by a DNA prime and inactivated virions-boost immunization strategy. Among the monoclonal antibodies, 3 (H50, H56, and H57) exhibited hemagglutination inhibition activity. Western blot analyses revealed that all the monoclonal antibodies reacted to the prokaryotically expressed HA1 of A/Chicken/Henan/01/2004(H5N1). The monoclonal antibodies were then used to characterize 10 avian influenza H5N1 viruses isolated from China during 2004 to 2007, by using the hemagglutination inhibition test and the antigen-capture enzyme-linked immunosorbent assay. The isolates could be divided into 4 different antigenic groups according to their responses to the monoclonal antibodies. The antigenic grouping of these 10 H5N1 isolates, using these antibodies, did not completely match their phylogenetic classification based on the hemagglutinin sequences. The results showed there were antigenic variations within the subclade 2.3.4 of H5N1, which is predominant in China.

Huit anticorps monoclonaux dirigés contre l’hémagglutinine A du virus influenza de la grippe aviaire (H1N5) ont été produits selon une stratégie d’immunisation utilisant une sensibilisation par l’ADN et un rappel par des virions inactivés. Parmi les anticorps monoclonaux obtenus, 3 d’entre eux (H50, H56 et H57) possédaient une activité inhibitrice de l’hémagglutination. Des analyses par buvardage Western ont révélé que tous les anticorps monoclonaux reconnaissaient le virus de la grippe aviaire H5N1 exprimé dans des cellules procaryotes. Les anticorps monoclonaux ont ensuite été utilisés pour caractériser 10 souches virales d’influenza aviaire H5N1 isolées en Chine entre 2004 et 2007, à l’aide d’un test d’inhibition de l’hémagglutination et d’un ELISA par capture d’antigène. Les isolats pouvaient être divisés en 4 groupes antigéniques différents selon leurs réponses respectives aux anticorps monoclonaux. Le regroupement antigénique de ces 10 isolats de H5N1 à l’aide de ces anticorps ne correspondait pas complètement à leur classification phylogénique basée sur la séquence de l’hémagglutinine. Les résultats montrent qu’il existe une variation antigénique dans le sous-clade 2,3.4 du H5N1 qui est prédominant en Chine.

Document Type: Research Article

Publication date: January 1, 2011

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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