1887

Journal of General Virology header logo

Volume 70, Issue 9

Propagation of Hepatitis A Virus in Hybrid Cell Lines Derived from Marmoset Liver and Vero Cells Free

Abstract

Summary

To establish monkey liver cell lines with a high susceptibility to hepatitis A virus (HAV), marmoset () liver cells were fused with Vero cells deficient in hypoxanthine-guanine phosphoribosyltransferase and the resulting hybrid cells were selected in HAT medium. Of four hybrid cell lines obtained (S. 1a/Ve-1 to -4), three (S. 1a/Ve-1, -3 and -4) were equally susceptible to HAV infection. When inoculated with a virus isolated from marmoset liver tissue (10% liver tissue extract) or a faecal virus (10% stool extract) from a human hepatitis A patient, all susceptible cell lines showed a significant elevation of viral antigen activity as seen in radioimmunoassay and/or immunofluorescent antibody assays, at 4 to 6 weeks post-infection (p.i.) with the liver-derived inoculum and at 6 to 8 weeks p.i. with the stool-derived inoculum. In S. 1a/Ve-1 cells, a representative of the susceptible hybrid cell lines, full adaptation of HAV (liver tissue virus concentrate) to cell culture was attained after four serial passages. Thereafter, the virus grew to a plateau titre of 10 TCID/ml at 7 days p.i. in a growth experiment. The infected cells showed no cytopathic effects but eventually a persistent infection was established when a saturated level of virus growth was reached.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): HAV , hybrid cells and propagation
© Society for General Microbiology 1989
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-70-9-2487
1989-09-01
2024-09-14
Loading full text...

Full text loading...

/deliver/fulltext/jgv/70/9/JV0700092487.html?itemId=/content/journal/jgv/10.1099/0022-1317-70-9-2487&mimeType=html&fmt=ahah

References

  1. Anderson D. A. 1987; Cytopathology, plaque assay, and heat inactivation of hepatitis A virus strain HM175. Journal of Medical Virology 22:35–44
     [Google Scholar]
  2. Binn L. N., Lemon S. M., Marchwicki R. H., Redfield R. R., Gates N. L., Bancroft W. H. 1984; Primary isolation and serial passage of hepatitis A virus strains in primate cell cultures. Journal of Clinical Microbiology 20:28–33
     [Google Scholar]
  3. Bradley D. W., Schable C. A., Mccaustland K. A., Cook E. H., Murphy B. L., Fields H. A., Ebert J. W., Wheeler C., Maynard J. E. 1984; Hepatitis A virus. Growth characteristics of in vivo and in vitro propagated wild and attenuated virus strains. Journal of Medical Virology 14:373–386
     [Google Scholar]
  4. Cromeans T., Sobsey M. D., Fields H. A. 1987; Development of a plaque assay for a cytopathic, rapidly replicating isolate of hepatitis A virus. Journal of Medical Virology 22:45–56
     [Google Scholar]
  5. Daemer R. J., Feinstone S. M., Gust I. D., Purcell R. H. 1981; Propagation of human hepatitis A virus in African green monkey kidney cell culture: primary isolation and serial passage. Infection and Immunity 32:388–393
     [Google Scholar]
  6. Deinhardt F., Deinhardt J. B. 1984; Animal models. In Hepatitis A185–204 Gerety R. J. Orlando & London: Academic Press;
     [Google Scholar]
  7. Flehmig B. 1980; Hepatitis A virus in cell culture. I. Propagation of different hepatitis A virus isolates in a fetal rhesus monkey kidney cell line (FrhK-4). Medical Microbiology and Immunology 168:239–248
     [Google Scholar]
  8. Flehmig B. 1981; Hepatitis A virus in cell culture. II. Growth characteristics of hepatitis A virus in FrhK-4/R cells. Medical Microbiology and Immunology 170:73–81
     [Google Scholar]
  9. Flehmig B., Vallbracht A., Wurster G. 1981; Hepatitis A virus in cell culture. III. Propagation of hepatitis A virus in human embryo kidney cells and human embryo fibroblast strains. Medical Microbiology and Immunology 170:83–89
     [Google Scholar]
  10. Gauss-müller V., Froesner G. G., Deinhardt F. 1981; Propagation of hepatitis A virus in human embryo fibroblasts. Journal of Medical Virology 7:233–239
     [Google Scholar]
  11. Karayiannis P., Enticott I. M., Moore D., Pignatelli M., Brenes F., Scheuer P. J., Thomas H. C. 1986; Hepatitis A virus replication in tamarins and host immune response in relation to pathogenesis of liver cell damage. Journal of Medical Virology 18:261–276
     [Google Scholar]
  12. Kojima H., Shibayama T., Sato A., Suzuki S., Ichida F., Hamada C. 1981; Propagation of human hepatitis A virus in conventional cell lines. Journal of Medical Virology 7:273–286
     [Google Scholar]
  13. Lemon S. M., Binn L. N., Marchwicki R. H. 1983; Radioimmunofocus assay for quantitation of hepatitis A virus in cell cultures. Journal of Clinical Microbiology 17:834–839
     [Google Scholar]
  14. Littlefield J. W. 1964; Three degrees of guanyl acid inosinic acid pyrophosphorylase deficiency in mouse fibroblasts. Nature,London 203:1142–1144
     [Google Scholar]
  15. Locarnini S. A., Coulepis A. G., Westaway E. G., Gust I. d. 1981; Restricted replication of human hepatitis A virus in cell culture. Intracellular biochemical studies. Journal of Virology 37:216–225
     [Google Scholar]
  16. Nakamura T., Aoyama K., Ichihara A. 1981; Methods of preparation and primary culture of isolated hepatocytes. Protein, Nucleic Acid and Enzyme 24:55–76
     [Google Scholar]
  17. Provost P. J. 1984; In vitro propagation of hepatitis A virus. In Hepatitis A245–261 Gerety R. J. London: Academic Press;
     [Google Scholar]
  18. Provost P. J., Hilleman M. R. 1979; Propagation of human hepatitis A virus in cell culture in vitro. Proceedings of the Society for Experimental Biology and Medicine 160:213–221
     [Google Scholar]
  19. Provost P. J., Mcaleer W. J., Hillemann M. R. 1982; In vitro cultivation of hepatitis A virus. In Viral Hepatitis21–30 Szmuness W., Alter H. J., Maynard J. E. Philadelphia: Franklin Institute Press;
     [Google Scholar]
  20. Purcell R. H., Feinstone S. M., Ticehurst J. R., Daemer R. J., Baroudy B. M. 1984; Hepatitis A virus. In Viral Hepatitis and Liver Disease9–22 Vyas G. N., Dienstag J. L., Hoofnagle J. H. Orlando: Grune & Stratton;
     [Google Scholar]
  21. Reed L. J., Muench H. 1938; A simple method of estimating fifty per cent endpoints. American Journal of Hygiene 27:493–197
     [Google Scholar]
  22. Shibayama T., Kojima H., Ashida M., Hirose S., Sato A., Kamimura T., Hamada C., Shimizu S., Ichida F. 1985; Localization of hepatitis A virus in marmoset liver tissue during the acute phase of experimental infection. Gastroenterologia Japonica 20:564–572
     [Google Scholar]
  23. Vallbracht A., Hofmann l., Wurster K. G., Flehmig B. 1984; Persistent infection of human fibroblasts by hepatitis A virus. Journal of General Virology 65:609–615
     [Google Scholar]
  24. Venuti A., Di russo C., Del grosso N., Patti A.-M., Ruggeri F., De stasio P. R., Martiniello M. G., Pagnotti P., Degener A. M., Middulla M., Pana A., Perez-bercoff R. 1985; Isolation and molecular cloning of a fast-growing strain of human hepatitis A virus from its double-stranded replicative form. Journal of Virology 56:579–588
     [Google Scholar]
  25. Yasumura Y., Kawakita A. 1963; Study of SV40 by tissue cultures. Nippon rinsho 21:1201–1219
     [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-70-9-2487
Loading
Propagation of Hepatitis A Virus in Hybrid Cell Lines Derived from Marmoset Liver and Vero Cells
J. Gen. Virol. 70, 2487 (1989); https://doi.org/10.1099/0022-1317-70-9-2487
/content/journal/jgv/10.1099/0022-1317-70-9-2487
/content/journal/jgv/10.1099/0022-1317-70-9-2487
Loading

Data & Media loading...

Most cited 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