1887

Abstract

Summary

To identify further the surface proteins of the native virus, hepatitis B virus (HBV) particles purified from HBe antigen (Ag)-positive human sera were used as immunogens to produce murine monoclonal antibody (MAb)-secreting hybridomas. The specific binding of antibodies to the HBV envelope () proteins was determined in indirect radioimmunoassay and by Western blot analysis. Six MAbs directed against major hepatitis B surface antigen (HBsAg) recognized conformational epitopes on S proteins (P24/GP27). Three preS2-specific MAbs reacted with the middle proteins (GP33/GP36) in the 22 nm HBsAg spherical particles. One MAb, F222, was found to react specifically with the two very large (VL) HBV surface proteins with 54K and 66K. The epitope recognized by F222 was located on the protruding N terminus which, in the assembled virus particles, was readily split off by trypsin or V8 protease treatment. The presence of these VL proteins appeared to correspond to the presence of the large proteins (P39/GP42). The data described here indicate that F222 probably recognized an assembled topographic site which could be involved in virus entry into hepatocytes. Moreover, our results suggest that the preS-coded part of the HBV proteins, which is sensitive to proteases , could be unstable and stabilized by immunoglobulins.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-68-11-2759
1987-11-01
2022-08-10
Loading full text...

Full text loading...

/deliver/fulltext/jgv/68/11/JV0680112759.html?itemId=/content/journal/jgv/10.1099/0022-1317-68-11-2759&mimeType=html&fmt=ahah

References

  1. Alberti A., Pontisso P., Realdi G. 1981; Changes in hepatitis B virus DNA-polymerase activity in patients with chronic infection. Journal of Medical Virology 8:223–229
    [Google Scholar]
  2. Budkowska A., Riottot M. M., Dubreuil P., Lazizi Y., Brechot C., Sobczak E., Petit M. A., Pillot J. 1986; Monoclonal antibody recognizing preS2 epitope of hepatitis B virus: characterization of preS2 epitope and anti-preS2 antibody. Journal of Medical Virology 20:111–125
    [Google Scholar]
  3. Budkowska A., Dubreuil P., Riottot M. M., Briantals M. J., Pillot J. 1987; A monoclonal antibody enzyme immunoassay for the detection of epitopes encoded by the preS2-region of the hepatitis B virus genome. Journal of Immunological Methods 97:77–85
    [Google Scholar]
  4. Burnette W. N. 1981; “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated Protein A. Analytical Biochemistry 112:195–203
    [Google Scholar]
  5. Cheng K. c., Smith G. L., Moss B. 1986; Hepatitis B virus large surface protein is not secreted but is immunogenic when selectively expressed by recombinant vaccinia virus. Journal of Virology 60:337–344
    [Google Scholar]
  6. Heermann K. H., Goldmann U., Schwartz W., Seyffarth T., Baumgarten H., Gerlich W. 1984; Large surface proteins of hepatitis B virus containing the preS-sequence. Journal of Virology 52:396–402
    [Google Scholar]
  7. Irshad M., Gandhi B. M., Chawla T. C., Acharya S. K., Joshi Y. K., Tandon B. N. 1987; Studies OnHBsAg binding with polymerized human serum albumin by ELISA. Journal of Virological Methods 16:75–85
    [Google Scholar]
  8. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  9. Machida A., Kishimoto S., Ohnuma H, Ito Y., Miyamoto H., Funatsu G., Oda K., Usuda S., Togami S., Nakamura T., Miyakawa Y., Mayumi M. 1984; A polypeptide containing 55 amino acid residues coded by the preS-region of hepatitis B surface antigen polypeptide (P31) with the receptor for polymerized human as well as chimpanzee albumins. Gastroenterology 86:910–918
    [Google Scholar]
  10. Michel M. L., Pontisso P., Sobczak E., Malpiece Y., Streeck R. E., Tiollais P. 1984; Synthesis in animal cells of hepatitis B 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]
  11. Mishiro S., Imai M., Takahashi K., Machida A., Gotanda T., Miyakawa Y., Mayumi M. 1980; A 49000-dalton polypeptide bearing all antigenic determinants and full immunogenicity of 22 nm hepatitis B surface antigen particles. Journal of Immunology 124:1589–1593
    [Google Scholar]
  12. Neurath A. R., Kent S. B. H., Strick N., Taylor P., Stevens C. E. 1985; Hepatitis B virus contains preS gene-encoded domains. Nature; London: 315154–157
    [Google Scholar]
  13. Neurath A. R., Adamowicz P., Kent S. B. H., Riottot M. M., Strick N., Parker K., Offensperger W., Petit M. A., Wahl S., Budkowska A., Girard M., Pillot J. 1986a; Characterization of monoclonal antibodies specific for the preS2 region of the hepatitis B virus envelope protein. Molecular Immunology 23:991–997
    [Google Scholar]
  14. Neurath A. R., Kent S. B. H., Strick N., Parker K. 1986b; Identification and chemical synthesis of a host cell receptor binding site on hepatitis B virus. Cell 46:429–436
    [Google Scholar]
  15. Neurath A. R., Kent S. B. H., Strick N., Parker K., Courouce A. M., Riottot M. M., Petit M. A., Budkowska A., Girard M., Pillot J. 1987; Antibodies to synthetic peptides from the pre-S1 and pre-S2 regions of one subtype of the hepatitis B virus (HBV) envelope protein recognize all HBV subtypes. Molecular Immunology 24:975–980
    [Google Scholar]
  16. Örvell C., Norrby E. 1985; Antigenic structure of paramyxoviruses. In Immunochemistry of Viruses. The Basis for Serodiagnosis and Vaccines pp. 241–264 Van Regenmortel M. H. V., Neurath A. R. Edited by New York: Elsevier;
    [Google Scholar]
  17. Petit M. A., Capel F., Pillot J. 1985; Demonstration of a firm association between hepatitis B surface antigen proteins bearing polymerized human albumin binding sites and core-specific determinants in serum hepatitis B viral particles. Molecular Immunology 22:1279–1287
    [Google Scholar]
  18. Petit M. A., Maillard P., Capel F., Pillot J. 1986; Immunochemical structure of the hepatitis B surface antigen vaccine. II. Analysis of antibody responses in human sera against the envelope proteins. Molecular Immunology 23:511–523
    [Google Scholar]
  19. Petit M. A., Capel F., Riottot M. M., Pillot J. 1987; Epitope analysis of hepatitis B virus envelope glycoprotein complex by monoclonal antibodies. In Viral Hepatitis and Liver Disease New York: Alan R. Liss; in press
    [Google Scholar]
  20. Scotto J., Hadchouel M., Hecy C., Yvart J., Tiollais P., Brechot C. 1983; Detection of hepatitis B virus DNA in serum by a simple spot hybridization technique: comparison with results for other viral markers. Hepatology 3:279–284
    [Google Scholar]
  21. Stibbe W., Gerlich W. H. 1982; Variable protein composition of hepatitis B surface antigen from different donors. Virology 123:436–442
    [Google Scholar]
  22. Takahashi K., Kishimoto S., Ohnuma H., Machida A., Takai E., Tsuda F., Miyamoto H., Tanaka T., Marsushita K., Oda K., Miyakawa Y., Mayumi M. 1986; Polypeptides coded for by the region preS and gene S of hepatitis B virus DNA with the receptor for polymerized human serum albumin: expression on hepatitis B particles produced in the HBeAg or anti-HBe phase of hepatitis B virus infection. Journal of Immunology 136:3467–3472
    [Google Scholar]
  23. Tiollais P., Pourcel C., Dejean A. 1985; The hepatitis B virus. Nature; London: 317489–495
    [Google Scholar]
  24. Trevisan A., Gudat F., Guggenheim R., Krey G., Duermueller U., Lueond G., Dueggelin M., Landmann J., Tondelli P., Bianchi L. 1981; Demonstration of albumin receptors on isolated human hepatocytes by light and scanning electron microscopy. Hepatology 2:832
    [Google Scholar]
  25. Yewdell J. W., Gerhard W. 1981; Antigenic characterization of viruses by monoclonal antibodies. Annual Review of Microbiology 35:185–206
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-68-11-2759
Loading
/content/journal/jgv/10.1099/0022-1317-68-11-2759
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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