1887

Abstract

An isolate of SARS coronavirus (strain 2003VA2774) was obtained from a patient and used to infect Vero E6 cells. The replication cycle of the virus was followed from 1 to 30 h post-infection (p.i.). It was surprising to observe the swift growth of this human virus in Vero cells. Within the first hour of infection, the most obvious ultrastructural change was the proliferation of the Golgi complexes and related vesicles accompanied by swelling of some of the -Golgi sacs. Extracellular virus particles were present by 5 h p.i. in about 5 % of the cells and this increased dramatically to about 30 % of the cell population within an hour (6 h p.i.). Swollen Golgi sacs contained virus nucleocapsids at different stages of maturation. These virus precursors were also in large vacuoles and in close association with membrane whorls. The membrane whorls could be the replication complexes, since they appeared rather early in the replication cycle. As infection progressed from 12 to 21 h p.i., the cytoplasm of the infected cells was filled with numerous large, smooth-membraned vacuoles containing a mixture of mature virus and spherical cores. Several of these vacuoles were close to the cell periphery, ready to export out the mature progeny virus particles via exocytosis. By 24 to 30 h p.i., crystalline arrays of the extracellular virus particles were seen commonly at the cell surface.

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2003-12-01
2024-12-07
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References

  1. Chaloner-Larsson G., Johnson-Lussenburg C. M. 1981; Establishment and maintenance of a persistent infection of L123 cells by human coronavirus strain 299E. Arch Virol 69:117–129
    [Google Scholar]
  2. David-Ferreira J. F., Manaker R. A. 1965; An electron microscope study of the development of a mouse hepatitis virus in tissue culture cells. J Cell Biol 24:57–78
    [Google Scholar]
  3. Drosten C., Gunther S., Preiser W. 23 other authors 2003; Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med 348:1967–1976
    [Google Scholar]
  4. Fleming J. O., Trousdale M. D., Stohlman S. A., Weiner L. P. 1987; Pathogenic characteristics of neutralization-resistant variants of JHM coronavirus (MHV-4). Adv Exp Med Biol 218:333–342
    [Google Scholar]
  5. Fleming J. O., el-Zaatari F. A., Gilmore W., Berne J. D., Burks J. S., Stohlman S. A., Tourtellotte W. W., Weiner L. P. 1988; Antigenic assessment of coronaviruses isolated from patients with multiple sclerosis. Arch Neurol 45:629–633
    [Google Scholar]
  6. Frana M. F., Behnke J. N., Sturman L. S., Holmes K. V. 1985; Proteolytic cleavage of the E2 glycoprotein of murine coronavirus: host-dependent differences in proteolytic cleavage and cell fusion. J Virol 56:912–920
    [Google Scholar]
  7. Holmes K. V. 1990; Coronaviridae and their replication. In Virology , 2nd edition. pp  841–856 Edited by Fields B. N., Knipe D. M., Howley P. M., Chanock R. M., Monath T. P., Melnick J. L., Roizman B., Straus S. E. New York: Raven Press;
    [Google Scholar]
  8. Holmes K. V., Behnke J. N. 1981; Evolution of a coronavirus during persistent infection in vitro . Adv Exp Med Biol 142:287–299
    [Google Scholar]
  9. Holmes K. V., Doller E. W., Behnke J. N. 1981a; Analysis of the functions of coronavirus glycoproteins by differential inhibition of synthesis with tunicamycin. Adv Exp Med Biol 142:133–142
    [Google Scholar]
  10. Holmes K. V., Doller E. W., Sturman L. S. 1981b; Tunicamycin resistant glycosylation of coronavirus glycoprotein: demonstration of a novel type of viral glycoprotein. Virology 115:334–344
    [Google Scholar]
  11. Holmes K. V., Frana M. F., Robbins S. G., Sturman L. S. 1984; Coronavirus maturation. Adv Exp Med Biol 173:37–52
    [Google Scholar]
  12. Ksiazek T. G., Erdman D., Goldsmith C. S. 23 other authors 2003; A novel coronavirus associated with severe acute respiratory syndrome. N Engl J Med 348:1953–1966
    [Google Scholar]
  13. Lamontagne L. M., Dupuy J. M. 1984; Persistent infection with mouse hepatitis virus 3 in mouse lymphoid cell lines. Infect Immun 44:716–723
    [Google Scholar]
  14. Luby J. P., Clinton R., Kurtz S. 1999; Adaptation of human enteric coronavirus to growth in cell lines. J Clin Virol 12:43–51
    [Google Scholar]
  15. Lucas A., Coulter M., Anderson R., Dales S., Flintoff W. 1978; In vivo and in vitro models of demyelinating diseases II. Persistence and host-regulated thermosensitivity in cells of neural derivation infected with mouse hepatitis and measles viruses. Virology 88:325–337
    [Google Scholar]
  16. Marra M. A., Jones S. J., Astell C. R. 56 other authors 2003; The genome sequence of the SARS-associated coronavirus. Science 300:1399–1404
    [Google Scholar]
  17. Ng M.-L., Tan S.-H., See E.-E., Ooi E.-E., Ling A.-E. 2003; Entry and early events of severe acute respiratory syndrome coronavirus. J Med Virol 71:323–331
    [Google Scholar]
  18. Oshiro L. S. 1973; Coronaviruses. . In Ultrastructure of Animal Viruses and Bacteriophages: an Atlas pp  331–343 Edited by Dalton A. J., Haguenau F. Orlando: Academic Press;
    [Google Scholar]
  19. Rota P. A., Oberste M. S., Monroe S. S. 32 other authors 2003; Characterization of a novel coronavirus associated with severe acute respiratory syndrome. Science 300:1394–1399
    [Google Scholar]
  20. Ruan Y. J., Wei C. L., Ling A.-E. 17 other authors 2003; Comparative full length genome sequence analysis of 14 SARS coronavirus isolates and common mutations associated with putative origins of infection. Lancet 361:1779–1785 erratum 361, 1832
    [Google Scholar]
  21. Sturman L. S., Takemoto K. K. 1972; Enhanced growth of a murine coronavirus in transformed mouse cells. Infect Immun 6:501–507
    [Google Scholar]
  22. Sturman L. S., Holmes K. V. 1983; The molecular biology of coronaviruses. Adv Virus Res 28:35–112
    [Google Scholar]
  23. Sturman L. S., Holmes K. V., Behnke J. 1980; Isolation of coronavirus envelope glycoproteins and interaction with the viral nucleocapsid. J Virol 33:449–462
    [Google Scholar]
  24. Sussman M. A., Fleming J. O., Allen H., Stohlman S. A. 1987; Immune mediated clearance of JHM virus from the central nervous system. Adv Exp Med Biol 218:399–410
    [Google Scholar]
  25. Tooze J., Tooze S. A. 1985; Infection of AtT20 murine pituitary tumor cells by mouse hepatitis virus strain A59: virus budding is restricted to the Golgi region. Eur J Cell Biol 37:203–212
    [Google Scholar]
  26. Tooze J., Tooze S. A., Warren G. 1984; Replication of coronavirus MHV-A59 in sac− cells: determination of the first site of budding of progeny virions. Eur J Cell Biol 33:281–293
    [Google Scholar]
  27. Wege H., Siddell S., ter Meulen V. 1982; The biology and pathogenesis of coronaviruses. Curr Top Microbiol Immunol 99:165–200
    [Google Scholar]
  28. Wilhelmsen K. C., Leibowitz J. L., Bond C. W., Robb J. A. 1981; The replication of murine coronaviruses in enucleated cells. Virology 110:225–230
    [Google Scholar]
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