1887

Abstract

SUMMARY

Mouse hepatocytes were isolated by collagenase perfusion, maintained in non-proliferating monolayer culture and shown to retain liver cell function as judged by gluconeogenesis for 15 to 18 h. Such cells could be infected with and support the replication of a virulent strain of ectromelia virus. Virus antigen and characteristic cytoplasmic ‘B’-type poxvirus inclusion bodies were demonstrated by immunofluorescence in virtually all cells. By electron microscopy it was shown that ‘B’-type inclusions were the site of virus replication, and that the biogenesis of ectromelia virus and ultrastructural changes in hepatocytes were similar to those observed in infected mouse livers. Early cell rounding effects, a normal characteristic of poxvirus infections in tissue culture cells, were not seen in ectromelia-infected hepatocytes, although late degenerative changes did occur. Pulse-labelling of hepatocyte cultures with [S]methionine showed that ectromelia virus inhibited the rise in protein synthesis seen in controls and imposed a gradual decline in host protein synthesis to an extent and at a rate significantly different from that in mouse L929 cells. Gluconeogenesis was inhibited by ectromelia virus infection of hepatocytes.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-66-10-2171
1985-10-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/66/10/JV0660102171.html?itemId=/content/journal/jgv/10.1099/0022-1317-66-10-2171&mimeType=html&fmt=ahah

References

  1. Arnheiter H. 1980; Primary monolayer culture of adult mouse hepatocytes - a model for the study of hepatotropic viruses. Archives of Virology 63:11–22
    [Google Scholar]
  2. Arnheiter H., Baechi T., Haller O. 1982; Adult mouse hepatocytes in primary monolayer culture express genetic resistance to mouse hepatitis virus type 3. Journal of Immunology 129:1275–1281
    [Google Scholar]
  3. Assimacopoulos-Jeannet F., Exton J. H., Jeanrenaud B. 1973; Control of gluconeogenesis and glycogenolysis in perfused livers of normal mice. American Journal of Physiology 225:25–32
    [Google Scholar]
  4. Bablanian R. 1984; Pox virus cytopathogenicity: effects on cellular macromolecular synthesis. In Comprehensive Virology vol 19: pp 391–429 Edited by Fraenkel-conrat H., Wagner R. R. New York: Plenum Press;
    [Google Scholar]
  5. Birkbeck T. H., Stephen J. 1970; Specific removal of host-cell or vaccinia-virus antigens from extracts of infected cells by polyvalent disulphide-linked immunosorbents. Journal of General Virology 8:133–142
    [Google Scholar]
  6. Blanden R. V. 1970; Mechanisms of recovery from a generalized viral infection: mousepox. I. The effects of anti-thymocyte serum. Journal of Experimental Medicine 132:1035–1054
    [Google Scholar]
  7. Burrows R. B., Davidson P. F. 1982; Protein catabolism in cultures of hepatocytes derived from mice of various ages. Mechanisms of Aging and Development 19:85–94
    [Google Scholar]
  8. Fenner F. 1949; Mouse-pox (infectious ectromelia of mice): a review. Journal of Immunology 63:341–373
    [Google Scholar]
  9. Fraenkel-Conrat H., Wagner R. R. (editors) 1984 Virus Cytopathology. Comprehensive Virology vol. 19 New York: Plenum Press;
    [Google Scholar]
  10. Fukuda J., Kurate T., Yamamoto A., Yamaguch K. 1983; Morphological and physiological studies on cultured nerve cells from guinea pigs infected with herpes simplex virus in vivo. Brain Research 262:79–89
    [Google Scholar]
  11. Hanson R. W., Mehlman M. A. 1976 Gluconeogenesis: Its Regulation in Mammalian Species New York: John Wiley;
    [Google Scholar]
  12. Harper L., Bedson H. S., Buchan A. 1979; Identification of orthopox-virus by polyacrylamide gel electrophoresis of intracellular polypeptides. Virology 93:435–444
    [Google Scholar]
  13. Ichihashi Y., Matsumoto S., Dales S. 1971; Biogenesis of poxviruses; role of A-type inclusions and host cell membranes in virus dissemination. Virology 46:507–532
    [Google Scholar]
  14. Isom H. C. 1980; DNA synthesis in isolated hepatocytes infected with herpes viruses. Virology 103:199–216
    [Google Scholar]
  15. Jen G., Thach R. E. 1982; Inhibition of host translation in encephalomyocarditis virus-infected L cells: a novel mechanism. Journal of Virology 43:250–261
    [Google Scholar]
  16. Jen G., Detjen B. M., Thach R. E. 1980; Shut off of HeLa cell protein synthesis by encephalomyocarditis virus and poliovirus: a comparative study. Journal of Virology 35:150–156
    [Google Scholar]
  17. Kawamura S., Taguchi F., Ishida T., Nakayama M., Fujiwara K. 1983; Growth of Tyzzer’s organism in primary cultures of adult mouse hepatocytes. Journal of General Microbiology 129:277–283
    [Google Scholar]
  18. Kirn A., Keller F. 1983; How viruses may overcome non-specific defences in the host. Philosophical Transactions of the Royal Society B 303:115–122
    [Google Scholar]
  19. Kirn A., Gut J. -P., Bingen A., Steffan A. M. 1983; Murine hepatitis induced by frog virus 3: a model for studying the effect of Sinusoidal cell damage on the liver. Hepatology 3:105–111
    [Google Scholar]
  20. Klaunig J. E., Goldblatt P. J., Hinton D. E., Lipsky M. M., Chacko J., Trump B. F. 1981a; Mouse liver cell culture. I. Hepatocyte isolation. In Vitro 17:913–925
    [Google Scholar]
  21. Klaunig J. E., Goldblatt P. J., Hinton D. E., Lipsky M. M., Chacko J., Trump B. F. 1981b; Mouse liver Cell culture. II. Primary culture. In Vitro 17:926–934
    [Google Scholar]
  22. Leduc F. H., Bernhard W. 1962; Electron microscopy study of mouse liver infected by ectromelia virus. Journal of Ultrastructure Research 6:466–488
    [Google Scholar]
  23. Maslansky C. J., Williams G. H. 1982; Primary cultures and the levels of cytochrome P450 in hepatocytes from mouse, rat, hamster and rabbit liver. In Vitro 18:683–693
    [Google Scholar]
  24. Michalopoulos G., Sattler G. L., Pitot H. C. 1978; Hormonal regulation and the effects of glucose on tyrosine aminotransferase activity in adult rat hepatocytes cultured on floating collagen membranes. Cancer Research 38:1550–1555
    [Google Scholar]
  25. Mims C. A. 1964; Aspects of the pathogenesis of virus disease. Bacteriological Reviews 28:30–71
    [Google Scholar]
  26. Mogensen S. C. 1979; Role of macrophages in natural resistance to virus infections. Microbiological Reviews 43:1–26
    [Google Scholar]
  27. Moss B. 1974; Reproduction of poxviruses. In Comprehensive Virology vol 3: pp 405–474 Edited by Fraenkel-Conrat H., Wagner R. R. New York: Plenum Press;
    [Google Scholar]
  28. Nossal G. J. V., De Burgh P. M. 1954; Ectromelia virus multiplication in mouse liver. Journal of General Microbiology 10:345–352
    [Google Scholar]
  29. O’Farrell P. H. 1975; High resolution two-dimensional electrophoresis of proteins. Journal of Biological Chemistry 250:4007–4021
    [Google Scholar]
  30. Oldstone M. B. A., Fujinami R. S. 1982; Virus persistence and avoidance of immune surveillance: how measles can be induced to persist in cells, escape immune assault and injure tissues. In Virus Persistence pp 185–202 Symposia of the Society for General Microbiology vol 33 Edited by Mahy B. W. J., Minson A. C., Darby G. K. Cambridge: Cambridge University Press;
    [Google Scholar]
  31. Oldstone M. B. A., Sinha Y. N., Blount P., Tishon A., Rodriguez M., Von Wedel R., Lampert P. W. 1982; Virus-induced alterations in homeostasis: alterations in differentiated functions of infected cells in vivo. Science 218:1125–1127
    [Google Scholar]
  32. Otto M. J., Lucas-Lenard J. 1980; The influence of host cell on the inhibition of virus protein synthesis in cells doubly infected with vesicular stomatitis virus and mengovirus. Journal of General Virology 50:293–307
    [Google Scholar]
  33. Ozaki Y., Higashi N. 1959; 3-a. Studies on the growth of viruses of ectromelia and vaccinia in strain L cells and HeLa cells. Annual Report, Institute of Virus Research, Kyoto University, Series B 2:65–113
    [Google Scholar]
  34. Page D. T., Garvey J. S. 1979; Isolation and characterization of hepatocytes and Kupffer cells. Journal of Immunological Methods 27:159–173
    [Google Scholar]
  35. Porter D. D., Muck K. B., Prince G. A. 1980; The age dependence of respiratory syncytial virus growth in ferret lung can be shown in organ and monolayer culture. Clinical Immunology and Immunopathology 15:415–423
    [Google Scholar]
  36. Rodgers B. C., Mims C. A. 1981; Interaction of influenza virus with mouse macrophages. Infection and Immunity 31:751–757
    [Google Scholar]
  37. Rodgers B. C., Mims C. A. 1982; Influenza virus replication in human alveolar macrophages. Journal of Medical Virology 9:177–184
    [Google Scholar]
  38. Seglen P. O. 1976; Preparation of isolated rat liver cells. Methods in Cell Biology 13:29–83
    [Google Scholar]
  39. Seglen P. O., Solheirn A. E., Grinde B., Gordon P. B., Schwarze P. E., Gjessing R., Poli A. 1980; Amino acid control of protein synthesis and degradation in isolated rat hepatocytes. Annals of the New York Academy of Sciences 349:1–17
    [Google Scholar]
  40. Shatkin A. J. 1983; Molecular mechanisms of virus-mediated cytopathology. Philosophical Transactions of the Royal Society B 303:167–178
    [Google Scholar]
  41. Smith C. I., Hammarstrom L. 1979; Detection of albumin-secreting hepatocytes at the single cell level by a protein A plaque assay. European Journal of Immunology 9:570–571
    [Google Scholar]
  42. Taguchi F., Kawamura S., Fujiwara K. 1983; Replication of mouse hepatitis viruses with high and low virulence in cultured hepatocytes. Infection and Immunity 39:955–959
    [Google Scholar]
  43. Yamada S., Otto S. P., Kennedy L. D., Whayne T. F. 1980; The effects of dexamethasone on metabolic activity of hepatocytes in primary monolayer culture. In Vitro 16:559–570
    [Google Scholar]
  44. Zaleski J., Bryla J. 1977; Effects of oleate, palmitate and octanoate on gluconeogenesis in isolated rabbit liver cells. Archives of Biochemistry and Biophysics 183:553–562
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-66-10-2171
Loading
/content/journal/jgv/10.1099/0022-1317-66-10-2171
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