1887

Abstract

We used the polymerase chain reaction to amplify the HA1 coding region of influenza A (H1N1) viruses present in clinical material from recent cases of influenza in the U.K. Previously, we have demonstrated that isolation of human influenza viruses in embryonated hens’ eggs selects variants which have amino acid substitutions in their haemagglutinin (HA) clustering around the receptor-binding site. Such egg-selected variants are often antigenically distinct from each other and from corresponding viruses isolated on mammalian cells. Since in general the virus used for vaccine production is an egg-adapted virus, it is important to determine the extent to which these variants are present in the natural virus which causes disease in man. To achieve this, amplified products from clinical material were cloned and many individual clones sequenced. Our results indicate that the HA of the naturally occurring virus is relatively homogeneous and represented by virus isolated in the laboratory on MDCK cells, whereas the variants isolated in eggs are present only at low levels in clinical material.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-11-2671
1991-11-01
2024-11-07
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/11/JV0720112671.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-11-2671&mimeType=html&fmt=ahah

References

  1. Aytay S., Schulze I. T. 1991; Single amino acid substitutions in the hemagglutinin can alter the host range and receptor binding properties of H1 strains of influenza A virus. Journal of Virology 65:3022–3028
    [Google Scholar]
  2. Cox N. J., Black R. A., Kendal A. P. 1989; Pathways of evolution of influenza A (H1N1) viruses from 1977 to 1986 as determined by oligonucleotide mapping and sequencing studies. Journal of General Virology 70:299–313
    [Google Scholar]
  3. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Research 12:387–395
    [Google Scholar]
  4. Fields S., Winter G. 1981; Nucleotide sequence heterogeneity and sequence rearrangements in influenza virus cDNA. Gene 15:207–214
    [Google Scholar]
  5. Katz J. M., Webster R. G. 1988; Antigenic and structural characterization of multiple subpopulations of H3N2 influenza virus from an individual. Virology 165:446–456
    [Google Scholar]
  6. Katz J. M., Wang M., Webster R. G. 1990; Direct sequencing of the HA gene of influenza (H3N2) virus in original clinical samples reveals sequence identity with mammalian cell-grown virus. Journal of Virology 64:1808–1811
    [Google Scholar]
  7. Mums K. B., Faloona F. A. 1987; Specific synthesis of DNA in vitro via a polymerase-catalysed chain reaction. Methods of Enzymology 155:335–350
    [Google Scholar]
  8. Nobusawa E., Nakajima K. 1988; Amino acid substitution at position 226 of the hemagglutinin molecule of influenza (H1N1) virus affects receptor binding activity but not fusion activity. Virology 167:8–14
    [Google Scholar]
  9. Oxford J. S., Corcoran T., Knott R., Bates J., Bartolomei O., Major D., Newman R. W., Yates P., Robertson J., Webster R. G., Schild G. C. 1987; Serological studies with influenza A (H1N1) viruses cultivated in eggs or in a canine kidney cell line (MDCK). Bulletin of the World Health Organization 65:181–187
    [Google Scholar]
  10. Parvin J. D., Moscona A., Pan W. T., Leider J. M., Palese P. 1986; Measurement of the mutation rates of animal viruses: influenza A virus and poliovirus type 1. Journal of Virology 59:377–383
    [Google Scholar]
  11. Portner A., Webster R. G., Bean W. J. 1980; Similar frequencies of antigenic variants in Sendai, vesicular stomatitis, and influenza A viruses. Virology 104:235–238
    [Google Scholar]
  12. Rajakumar A., Swierkosz E. M., Schulze I. T. 1990; Sequence of an influenza virus hemagglutinin determined directly from a clinical sample. Proceedings of the National Academy of Sciences, U.S.A. 87:4154–4158
    [Google Scholar]
  13. Robertson J. S. 1987; Sequence analysis of the haemagglutinin of A/Taiwan/1/86, a new variant of human influenza A (H1N1) virus. Journal of General Virology 68:1205–1208
    [Google Scholar]
  14. Robertson J. S., Naeve C. W., Webster R. G., Bootman J. S., Newman R., Schild G. C. 1985; Alterations in the hemagglutinin associated with adaptation of influenza B virus to growth in eggs. Virology 143:166–174
    [Google Scholar]
  15. Robertson J. S., Bootman J. S., Newman R., Oxford J. S., Daniels R. S., Webster R. G., Schild G. C. 1987; Structural changes in the hemagglutinin which accompany egg adaptation of an influenza A (H1N1) virus. Virology 160:31–37
    [Google Scholar]
  16. Robertson J. S., Bootman J. S., Nicolson C., Major D., Robertson E. W., Wood J. M. 1990; The hemagglutinin of influenza B virus present in clinical material is a single species identical to that of mammalian cell-grown virus. Virology 179:35–40
    [Google Scholar]
  17. Rogers G. N., Paulson J. C., Daniels R. S., Skehel J. J., Wilson I. A., Wiley D. C. 1983; Single amino acid substitutions in influenza haemagglutinin change receptor binding specificity. Nature, London 304:76–78
    [Google Scholar]
  18. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Ehrlich H. A. 1988; Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
    [Google Scholar]
  19. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd. edn New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A. 74:5463–5467
    [Google Scholar]
  21. Staden R. 1982; Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. Nucleic Acids Research 10:4731–4751
    [Google Scholar]
  22. Tindall K. R., Kunkel T. A. 1988; Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase. Biochemistry 27:60086013
    [Google Scholar]
  23. Wang M., Katz J. M., Webster R. G. 1989; Extensive heterogeneity in the hemagglutinin of egg-grown influenza viruses from different patients. Virology 171:275–279
    [Google Scholar]
  24. Wilson I. A., Skehel J. J., Wiley D. C. 1981; Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3A resolution. Nature, London 289:366–373
    [Google Scholar]
  25. Wood J. M., Oxford J. S., Dunleavy U., Newman R. W., Major D., Robertson J. S. 1989; Influenza A (H1N1) vaccine efficacy in animal models is influenced by two amino acid substitutions in the hemagglutinin molecule. Virology 171:214–221
    [Google Scholar]
  26. Yates P. J., Bootman J. S., Robertson J. S. 1990; The antigenic structure of a human influenza A (H1N1) virus isolate grown exclusively in MDCK cells. Journal of General Virology 71:1683–1688
    [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-72-11-2671
Loading
/content/journal/jgv/10.1099/0022-1317-72-11-2671
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error