1887

Abstract

The degree of susceptibility of human hepatoma (HepG2) cells to direct hepatitis B virus (HBV) infection remains unknown. We previously observed a low level of Dane particle production and viral DNA replication after infection of HepG2 cells with serum-derived HBV. However, this culture system appeared to be affected by variations as human hepatocyte cultures. In the present study, HBV infection of HepG2 cells led to a significant increase in the secretion of three envelope antigens (HBsAg, preS2Ag and preSl Ag) at 4 days post-infection, and Northern blot analysis revealed the presence of both preSl (2·6 kb) and preS2/S (2·2 kb) transcripts. Expression of preSlAg and the corresponding viral RNA became undetectable on 21 days post-infection whereas the 2·2 kb RNA species persisted and was associated with secretion of subviral HBs particles expressing preS2-epitopes and banding between 30 and 35% sucrose. At 35 days post-infection (fifth passage), a sudden high level production of HBsAg and preSlAg was observed, followed by a massive cell death (90 %). A stable HBsAg-producing HepG2 cell line, designated HepG2-BV3, grew out of the surviving cells. HepG2- BV3 cells could integrate HBV DNA sequences and produce the three HBV surface antigens. Treatment with dexamethasone increased the HBsAg and preSlAg secretion. Such a HBsAg-producing HepG2 cell line obtained by HBV infection seems to mimick events that occur in the naturally occurring persistent chronic infection, and therefore may be an efficient model for studying the contribution of viral integration in the dysregulation of HBV and liver- specific genes expression.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-75-10-2681
1994-10-01
2022-01-24
Loading full text...

Full text loading...

/deliver/fulltext/jgv/75/10/JV0750102681.html?itemId=/content/journal/jgv/10.1099/0022-1317-75-10-2681&mimeType=html&fmt=ahah

References

  1. Aden D. P., Fogel A., Plotkin S., Damjanov I., Knowles B. B. 1979; Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell-line. Nature; London: 282615–616
    [Google Scholar]
  2. Alexander J. J., Bey E. M., Geddes E. W., Lecatsas G. 1976; Establishment of a continuously growing cell line from primary carcinoma of the liver. South African Medical Journal 50:2124–2128
    [Google Scholar]
  3. Bchini R., Capel F., Dauguet C., Dubanchet S., Petit M. A. 1990; In vitro infection of human hepatoma (HepG2) cells with hepatitis B virus. Journal of Virology 64:3025–3032
    [Google Scholar]
  4. Charnay P., Pourcel C., Fritsch A., Louise A., Tiollais P. 1979; Cloning in Escherichia coli and physical structure of hepatitis B virion DNA. Proceedings of the National Academy of Sciences, U.S.A 76:2222–2226
    [Google Scholar]
  5. Chisari F. V., Filippi P., Buras J., Mclachlan A., Popper H., Pinkert C. A., Palmiter R. D., Brinster R. L. 1987; Structural and pathological effects of synthesis of hepatitis B virus large envelope polypeptide in transgenic mice. Proceedings of the National Academy of Sciences, U.S.A 84:6909–6913
    [Google Scholar]
  6. Courtois G., Baumhueter S., Crabtree G. R. 1988; Purified hepatocyte nuclear factor 1 interacts with a family of hepatocyte-specific promoters. Proceedings of the National Academy of Sciences, U.S.A 85:7937–7941
    [Google Scholar]
  7. Dienes H. P., Gerlich W. H., Worsdorfer M., Gerken G., Bianchi L., Hess G., Zum Bushenfelde K. M. 1990; Hepatic expression patterns of the large and middle hepatitis B virus surface proteins in viremic and nonviremic chronic hepatitis B. Gastroenterology 98:1017–1023
    [Google Scholar]
  8. Farza H., Salmon A. M., Hadchouel M., Moreau J. L., Babinet C., Tiollais P., Pourcel C. 1987; Hepatitis B surface antigen gene expression is regulated by sex steroids and glucocorticoids in transgenic mice. Proceedings of the National Academy of Sciences, U.S.A 84:1187–1191
    [Google Scholar]
  9. Gripon P., Diot C., Theze N., Fourel I., Loreal O., Brechot C., Guguen-Guillouzo C. 1988; Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide. Journal of Virology 62:4136–4143
    [Google Scholar]
  10. Huang Z.-M., Yen T. S. B. 1993; Dysregulated surface gene expression from disrupted hepatitis B virus genomes. Journal of Virology 67:7032–7040
    [Google Scholar]
  11. Kekule A. S., Lauer U., Weisse L., Luber B., Hofschneider P. H. 1993; Hepatitis B virus transactivator HBx utilizes a tumour promoter signalling pathway. Nature; London: 361742–745
    [Google Scholar]
  12. Knowles B. B., Searls D. B., Aden D. P. 1984; Human hepatoma-derived cell lines. In Advances in Hepatitis Research pp. 196–202 Chisari F. V. Edited by New York: Masson Publishing;
    [Google Scholar]
  13. Lauer U., Weisse L., Lipp M., Hofschneider P. H., Kekule A. S. 1994; The hepatitis B virus PreS2/St transactivator utilizes AP-1 and other transcription factors for transactivation. Hepatology 19:23–31
    [Google Scholar]
  14. Mason D. Y., Sammons R. E. 1979; The labeled antigen method of immunoenzymatic staining. Journal of Histochemistry and Cytochemistry 27:832–840
    [Google Scholar]
  15. Nayersina R., Fowler P., Guilhot S., Missale G., Cerny A., Schlicht H.-J., Vitiello A., Chesnut R., Person J. L., Redeker A. G., Chisari F. V. 1993; HLA A2 restricted cytotoxic T lymphocyte responses to multiple hepatitis B surface antigen epitope during hepatitis B virus infection. Journal of Immunology 150:4659–4671
    [Google Scholar]
  16. Norder H., Hammas B., Lee S.-D., Bile K., Couroucé A.-M., Mushahwar I. K., Magnius L. O. 1993; Genetic relatedness of hepatitis B viral strains of diverse geographical origin and natural variations in the primary structure of the surface antigen. Journal of General Virology 74:1341–1348
    [Google Scholar]
  17. Ochiya T., Tsurimoto T., Ueda K., Okubo K., Shiozawa M., Matsubara K. 1989; An in vitro system for infection with hepatitis B virus that uses primary human fetal hepatocytes. Proceedings of the National Academy of Sciences, U.S.A 86:1875–1879
    [Google Scholar]
  18. Ou J. H., Rutter W. J. 1987; Regulation of secretion of the hepatitis B virus major surface antigen by the preSl protein. Journal of Virology 61:782–786
    [Google Scholar]
  19. Penna A., Chisari F. V., Bertoletti A., Missale G., Fowler P., Giuberti T., Fiaccadori F., Ferrari C. 1991; Cytotoxic T lymphocytes recognize an HLA-A2-restricted epitope within the hepatitis B virus nucleocapsid antigen. Journal of Experimental Medicine 174:1565–1570
    [Google Scholar]
  20. Petit M. A., Zoulim F., Capel F., Dubanchet S., Dauguet C., Trepo C. 1990; Variable expression of PreSl antigen in serum during chronic hepatitis B virus infection: an accurate marker for the level of hepatitis B virus replication. Hepatology 11:809–814
    [Google Scholar]
  21. Petit M. A., Dubanchet S., Capel F., Voet P., Dauguet C., Hauser P. 1991a; HepG2 cell binding activities of different hepatitis B virus isolates: inhibitory effect of anti-HBs and anti-PreS1 (21-47). Virology 180:483–491
    [Google Scholar]
  22. Petit M. A., Mabit H., Dubanchet S., Capel F. 1991b; Hepatitis B virus (HBV) envelope antigen accumulation in HBV nonproducer HepG2-infected cells. Life Science Advances 10:125–137
    [Google Scholar]
  23. Petit M. A., Strick N., Dubanchet S., Capel F., Neurath A. R. 1991c; Inhibitory activity of monoclonal antibody F35.25 on the interaction between hepatocytes (HepG2 cells) and preS1-specific ligands. Molecular Immunology 28:517–521
    [Google Scholar]
  24. Petit M. A., Capel F., Dubanchet S., Mabit H. 1992; PreS1-specific binding proteins as potential receptors for hepatitis B virus in human hepatocytes. Virology 187:211–222
    [Google Scholar]
  25. Sells M. A., Chen M. L., Acs G. 1987; Production of hepatitis B virus particles in HepG2 cells transfected with cloned hepatitis B virus DNA. Proceedings of the National Academy of Sciences, U.S.A 84:1005–1009
    [Google Scholar]
  26. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  27. Sureau C., Romet-Lemonne J. L., Mullins J. I., Essex M. 1986; Production of hepatitis B virus by a differentiated human hepatoma cell line after transfection with cloned circular HBV DNA. Cell 47:37–47
    [Google Scholar]
  28. Tsurimoto T., Gripon P., Fourel I., Hantz O., Trepo C., Guguen-Guillouzo C. 1987; Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA. Proceedings of the National Academy of Sciences, U.S.A 84:444–448
    [Google Scholar]
  29. Tur-Kaspa R., Laub O. 1990; Corticoids stimulate hepatitis B virus DNA, mRNA and protein production in a stable expression system. Journal of Hepatology 11:34–36
    [Google Scholar]
  30. Tur-Kaspa R., Burk R. D., Shaul Y., Shafritz D. A. 1986; Hepatitis B virus contains a glucocorticoid-responsive element. Proceedings of the National Academy of Sciences, U.S.A 83:1627–1631
    [Google Scholar]
  31. Yuasa T., Kajino K., Saito I., Miyamura T. 1991; Preferential expression of the large hepatitis B virus surface antigen gene by an adenovirus-hepatitis B virus recombinant. Journal of General Virology 72:1927–1934
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-75-10-2681
Loading
/content/journal/jgv/10.1099/0022-1317-75-10-2681
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