1887

Abstract

To elucidate the structure of the antigenic sites of avian H5 influenza virus haemagglutinin (HA) we analysed escape mutants of a mouse-adapted variant of the H5N2 strain A/Mallard/Pennsylvania/10218/84. A panel of five anti-H5 monoclonal antibodies (mAbs) was used to select 16 escape mutants. The mutants were tested by ELISA and haemagglutination inhibition with this panel of anti-H5 mAbs and the HA genes of the mutants were sequenced. The sequencing demonstrated that the amino acid changes were grouped in two antigenic sites. One corresponded to site A in the H3 HA. The other contained areas that are separated in the amino acid sequence but are topographically close in the three-dimensional structure and partially overlap in the reactions with mAbs. This site corresponds in part to site B in the H3 structure; it also includes a region not involved in site B that partially overlaps site Sa in the H1 HA and an antigenic area in H2 HA. Mutants with the amino acid change K152N, as well as those with the change D126N, showed reduced lethality in mice. The substitution D126N, creating a new glycosylation site, was accompanied by an increase in the sensitivity of the mutants to normal mouse serum inhibitors. Several amino acid changes in the H5 escape mutants occurred at the positions of reported changes in H2 drift variants. This coincidence suggests that the antigenic sites described and analysed here may be important for drift variation if H5 influenza virus ever appears as a pathogen circulating in humans.

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2002-10-01
2024-12-05
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References

  1. Alexander D. J., Lister S. A., Johnson M. J., Randall C. J., Thomas P. J. 1993; An outbreak of highly pathogenic avian influenza in turkeys in Great Britain in 1991. Veterinary Record 132:535–536
    [Google Scholar]
  2. Bean W. J., Kawaoka Y., Wood J. M., Pearson J. E., Webster R. G. 1985; Characterization of virulent and avirulent A/chicken/Pennsylvania/83 viruses: potential role of defective interfering RNAs in nature. Journal of Virology 54:151–160
    [Google Scholar]
  3. Caton A. J., Brownlee G. G., Yewdell J. M., Gerhard W. 1982; The antigenic structure of the influenza virus A/PR/8/34 hemagglutinin (H1 subtype). Cell 31:417–427
    [Google Scholar]
  4. Cauthen A. N., Swayne D. E., Schultz-Cherry S., Perdue M. L., Suarez D. L. 2000; Continued circulation in China of highly pathogenic avian influenza viruses encoding the hemagglutinin gene associated with the 1997 outbreak in poultry and humans. Journal of Virology 74:6592–6599
    [Google Scholar]
  5. Centers for Disease Control & Prevention 1998; Update: isolation of avian influenza A (H5N1) viruses from humans – Hong Kong, 1997–1998. Morbidity and Mortality Weekly Report 46:1245–1247
    [Google Scholar]
  6. Claas E. C. J., Osterhaus A. D. M. E., van Beek R., de Jong J. C., Rimmelzwaan G. F., Senne D. A., Krauss S., Shortridge K. F., Webster R. G. 1998; Human influenza A H5N1 virus related to a highly pathogenic avian influenza virus. Lancet 351:472–477
    [Google Scholar]
  7. Daniels R. S., Jeffries S., Yates P., Schild G. C., Rodgers G. N., Paulson J. C., Wharton S. A., Douglas A. R., Skehel J. J., Wiley D. C. 1987; The receptor-binding and membrane-fusion properties of influenza virus variants selected using anti-haemagglutinin monoclonal antibodies. EMBO Journal 6:1459–1465
    [Google Scholar]
  8. Gitelman A. K., Kaverin N. V., Kharotonenkov I. G., Rudneva I. A., Sklyanskaya E. I., Zhdanov V. M. 1986; Dissociation of the haemagglutination inhibition and the infectivity neutralization in the reactions of influenza A/USSR/90/77 (H1N1) virus variants with monoclonal antibodies. Journal of General Virology 67:2247–2251
    [Google Scholar]
  9. Ha Y., Stevens D. J., Skehel J. J., Wiley D. C. 2001; X-ray structures of H5 avian and H9 swine influenza virus hemagglutinins bound to avian and human receptor analogs. Proceedings of the National Academy of Sciences, USA 98:11181–11186
    [Google Scholar]
  10. Horimoto T., Rivera E., Pearson J., Senne D., Krauss S., Kawaoka Y., Webster R. G. 1995; Origin and molecular changes associated with emergence of a highly pathogenic H5N2 influenza virus in Mexico. Virology 213:223–230
    [Google Scholar]
  11. Laemmly U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  12. Murphy B. R., Webster R. G. 1990; Orthomyxoviruses. In Fields Virology pp 1091–1152 Edited by Fields B. N., Knipe D. M. New York: Raven Press;
    [Google Scholar]
  13. Nobusawa E., Aoyama T., Kato H., Suzuki Y., Tateno Y., Nakajima K. 1991; Comparison of complete amino acid sequences and receptor-binding properties among 13 serotypes of hemagglutinins of influenza A viruses. Virology 182:475–485
    [Google Scholar]
  14. Palmer D. F., Dowdle W. R., Coleman M. T., Schild G. C. 1975; Advanced Laboratory Techniques for Influenza Diagnosis. US Department of Health, Education and Welfare Immunology Series No. 6. Atlanta, GA: Center for Disease Control;
    [Google Scholar]
  15. Philpott M., Easterday B. C., Hinshaw V. S. 1989; Neutralizing epitopes of the H5 hemagglutinin from a virulent avian influenza virus and their relationship to pathogenicity. Journal of Virology 63:3453–3458
    [Google Scholar]
  16. Philpott M., Hioe C., Sheerar M., Hinshaw V. S. 1990; Hemagglutinin mutations related to attenuation and altered cell tropism of a virulent avian influenza A virus. Journal of Virology 64:2941–2947
    [Google Scholar]
  17. Reading P. C., Morey L. S., Crouch I. C., Anders M. E. 1997; Collectin-mediated antiviral host defence of the lung: evidence from influenza virus infection of mice. Journal of Virology 71:8204–8212
    [Google Scholar]
  18. Reed L. J., Muench H. 1932; A simple method for estimating 50% endpoints. American Journal of Hygiene 27:493–497
    [Google Scholar]
  19. Smirnov Y. A., Lipatov A. S., van Beek R., Gitelman A. K., Osterhaus A. D. M. E., Claas E. C. J. 2000; Characterization of adaptation of an avian influenza A (H5N2) virus to a mammalian host. Acta Virologica 44:1–8
    [Google Scholar]
  20. Smirnov Y. A., Kaverin N. V., Govorkova E. A., Lipatov A. S., Claas E. C. J., Makarova N. V., Gitelman A. K., Webster R. G., Lvov D. K. 2001; Cross-protection studies with H5 influenza viruses. In Options for the Control of Influenza IV, International Congress Series 1219 pp 767–773 Edited by Osterhaus A. D. M. E., Cox N., Hampson A. W. Amsterdam: Excerpta Medica;
    [Google Scholar]
  21. Suarez D. L., Perdue M. L., Cox N., Rowe T., Bender C., Huang J., Swayne D. E. 1998; Comparisons of highly virulent H5N1 influenza viruses isolated from humans and chickens from Hong Kong. Journal of Virology 72:6678–6688
    [Google Scholar]
  22. Swayne D. E. 1997; Pathobiology of H5N2 Mexican avian influenza viruses for chickens. Veterinary Pathology 34:557–567
    [Google Scholar]
  23. Swayne D. E., Perdue M. L., Garcia M., Rivera-Cruz E., Brugh M. 1997; Pathogenicity and diagnosis of H5N2 Mexican avian influenza viruses in chickens. Avian Diseases 41:335–346
    [Google Scholar]
  24. Tsuchiya E., Sugawara K., Hongo S., Matsuzaki Y., Muraki Y., Li Z.-N., Nakamura K. 2001; Antigenic structure of the haemagglutinin of human influenza A/H2N2 virus. Journal of General Virology 82:2475–2484
    [Google Scholar]
  25. Webster R. G., Laver W. G. 1980; Determination of the number of nonoverlapping antigenic areas on Hong Kong (H3N2) influenza virus hemagglutinin with monoclonal antibodies and the selection of variants with potential epidemiological significance. Virology 104:139–148
    [Google Scholar]
  26. Webster R. G., Bean W. J., Gorman O. T., Chambers T. M., Kawaoka Y. 1992; Evolution and ecology of influenza A viruses. Microbiology and Molecular Biology Reviews 56:152–179
    [Google Scholar]
  27. Webster R. G., Guan Y., Peiris M., Walker D., Krauss S., Zhou N. N., Govorkova E. A., Ellis T. M., Dyrting K. C., Sit T., Perez D. R., Shortridge K. F. 2002; Characterization of H5N1 influenza viruses that continue to circulate in geese in southeastern China. Journal of Virology 76:118–126
    [Google Scholar]
  28. Wiley D. C., Wilson I. A., Skehel J. J. 1981; Structural identification of the antibody-binding sites of Hong Kong influenza hemagglutinin and their involvement in antigenic variation. Nature 289:373–378
    [Google Scholar]
  29. Wilson I. A., Skehel J. J., Wiley D. C. 1981; Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 Å resolution. Nature 289:366–373
    [Google Scholar]
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