Antigenic characterization of H5N1 highly pathogenic avian influenza viruses isolated from poultry in India, 2006-2015
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Highly pathogenic avian influenza (HPAI) is a major health concern worldwide. In this study, we focused on antigenic analysis of HPAI H5N1 viruses isolated from poultry in India between 2006 and 2015 comprising 25 isolates from four phylogenetic clades 2.2 (1 isolate), 22.214.171.124 (1 isolate), 126.96.36.199a (17 isolates) and 188.8.131.52c (6 isolates). Seven H5N1 isolates from all four clades were selected for production of chicken antiserum, and antigenic analysis was carried out by hemagglutination inhibition (HI) assay. HI data indicated antigenic divergence (6-21 fold reduction in cross-reactivity) between the two recently emerged clades 184.108.40.206a and 220.127.116.11c. These two clades are highly divergent (21-128 fold reduction in HI titre) from the earlier clades 2.2 /18.104.22.168 isolated in India. However, a maximum of 2-fold and 4-fold reduction in cross-reactivity was observed within the isolates of homologous clades 22.214.171.124c and 126.96.36.199a, respectively. The molecular basis of inter-clade antigenic divergence was examined in the haemagglutinin (HA) antigenic sites of the H5N1 virus. Amino acid changes at 8 HA antigenic sites were observed between clades 188.8.131.52a and 184.108.40.206c, whereas 20-23 substitutions were observed between clades 220.127.116.11a/18.104.22.168c and 2.2/22.214.171.124. Therefore, a systematic analysis of antigenic drift of the contemporary field isolates is a pre-requisite for determining the suitable strain(s) for vaccine candidature.
KeywordsInfluenza Avian Influenza H5N1 Virus Avian Influenza Virus Haemagglutination Inhibition
We are thankful to the ICAR, New Delhi and Director, ICAR-NIHSAD, Bhopal for providing necessary facilities to carry out this work. Funding by DADF, MoA, GOI through CDDL grant is acknowledged. S. Bhat acknowledges Director, ICAR-Indian Veterinary Research Institute for financial support.
Compliance with ethical standards
This study was supported by CDDL Grant of DADF, Ministry of Agriculture and Farmers Welfare, Government of India. S. Bhat received junior research fellowship from ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh.
Conflicts of interest
The authors declare that they have no conflict of interest.
The animal experiment was carried out as per the guidelines of CPCSEA, Government of India with approval from Institutional Animal Ethics Committee (IAEC Approval No. 54/IAEC/HSADL/11).
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