1887

Abstract

Polyclonal antibodies directed against the preS2 and S domains of the woodchuck hepatitis virus (WHV) envelope proteins were prepared using synthetic peptides and fusion polypeptides as immunogens. They were tested by immunoblotting and immunoprecipitation of infected woodchuck sera and lysates of a eukaryotic cell line expressing WHV envelope proteins. Only one anti-peptide serum directed against the preS2 domain was reactive with WHV envelope proteins, recognizing the preS2 and preS1 proteins by their preS2 epitopes. With recombinant fusion proteins we generated several anti-S sera, which recognized all envelope proteins, and anti-preS2 antisera, which recognized the preS proteins. Results obtained with our antisera showed that sera of infected woodchucks lack the low glycosylated form (GP33) of the preS2 protein, unlike human hepatitis B virus.

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/content/journal/jgv/10.1099/0022-1317-72-2-421
1991-02-01
2022-01-20
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References

  1. Chou P. Y., Fasman G. D. 1978; The secondary structure of proteins from their amino acid sequence. Advances in Enzymology 47:45–148
    [Google Scholar]
  2. Galibert F., Chen T. N., Mandart E. 1982; Nucleotide sequence of a cloned woodchuck hepatitis virus genome. Comparison with HBV sequence. Journal of Virology 41:51–65
    [Google Scholar]
  3. Gazin C., Rigolet M., Briand J. P., Van Regenmortel M. H. V., Galibert F. 1986; Immunochemical detection of proteins related to the human c-myc exon 1. EMBO Journal S:2241–2250
    [Google Scholar]
  4. Gerin J. L., Alexander H., Shih J. W., Purcell R. H., Dapolito G., Engle R., Green N., Sutcliffe J. G., Shinnick T. M., Lerner R. A. 1983; Chemically synthesized peptides of hepatitis B surface antigen duplicate the d/y specificities and induce subtype-specific antibodies in chimpanzees. Proceedings of the National Academy of Sciences, U,. S,. A 80:2365–2369
    [Google Scholar]
  5. Heermann K. H., Goldmann U., Schwartz W., Seyffarth T., Baumgarten H., Gerlich W. H. 1984; Large surface proteins of hepatitis B virus containing the pre-S sequence. Journal of Virology 52:396–402
    [Google Scholar]
  6. Hopp T. P., Woods K. R. 1981; Prediction of protein antigenic determinants from amino acid sequences. Proceedings of the National Academy of Sciences, U,. S,. A 78:3824–3828
    [Google Scholar]
  7. Howard C. R. 1986; The biology of hepadnaviruses. Journal of General Virology 67:1215–1235
    [Google Scholar]
  8. Lerner R. A., Green N., Alexander H., Liu F. T., Sutcliffe J. G., Shinnick T. M. 1981; Chemically synthesized peptides predicted from the nucleotide sequence of the hepatitis B virus genome elicit antibodies reactive with the native envelope protein of Dane particles. Proceedings of the National Academy of Sciences, U,. S,. A 78:3403–3407
    [Google Scholar]
  9. Michel M. L., Pontisso P., Sobczak E., Malpiece Y., Streek R. E., Tiollais P. 1984; Synthesis in animal cells of hepatitis B virus surface antigen particles carrying a receptor for polymerized human serum albumin. Proceedings of the National Academy of Sciences, U,. S,. A 81:7708–7712
    [Google Scholar]
  10. Ou J. H., Rutter W. J. 1987; Regulation of secretion of the hepatitis B virus major surface antigen by the pre-S1 protein. Journal of Virology 61:782–786
    [Google Scholar]
  11. Pain D., Surolia A. 1981; Preparation of Protein A–peroxidase monoconjugate using a heterobifunctional reagent, and its use in enzyme immunoassays. Journal of Immunological Methods 40:219–230
    [Google Scholar]
  12. Petit M. A., Capel F., Riottot M. M., Dauget C., Pillot J. 1987; Antigenic mapping of the surface proteins of infectious hepatitis B virus particles. Journal of General Virology 68:2759–2767
    [Google Scholar]
  13. Pohl C. J., Cote P. J., Purcell R. H., Gerin J. L. 1986; Failure to detect polyalbumin-binding sites on the woodchuck hepatitis virus surface antigen: implications for the pathogenesis of hepatitis B virus in humans. Journal of Virology 60:943–949
    [Google Scholar]
  14. Remaut E., Stanssens P., Fiers W. 1981; Plasmid vectors for high efficiency expression controlled by the PL promoter of coliphage lambda. Gene 15:81–93
    [Google Scholar]
  15. Saiki R. K., Gelfand D. H., Stoffel S., Scharf J., Higushi R., Horn G. T., Mums K. B., Erlich H. A. 1988; Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:488–491
    [Google Scholar]
  16. Schaeffer E., Sninsky J. J. 1984; Predicted secondary structure similarity in the absence of primary amino acid sequence homology: hepatitis B virus open reading frames. Proceedings of the National Academy of Sciences, U,. S,. A. 81:2902–2906
    [Google Scholar]
  17. Schaeffer E., Snyder R. L., Sninsky J. J. 1986; Identification and localization of preS-encoded polypeptides from woodchuck and ground squirrel hepatitis viruses. Journal of Virology 57:173–182
    [Google Scholar]
  18. Urlaub G., Chasin L. A. 1980; Isolation of Chinese hamster cell mutants deficient in dihydrofolate reductase activity. Proceedings of the National Academy of Sciences, U,. S,. A 77:4216–4220
    [Google Scholar]
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