1887

Abstract

SUMMARY: Less than a tenth of the pock-forming particles of the HFEM strain of herpes virus are able to initiate infection in HeLa cells, even after passage in these cells. The pock-forming virus attaches firmly to the cells but most of it remains susceptible to antiserum and presumably does not penetrate the cell. A small proportion of virus initiates infection in the HeLa cells, and, after exposure to more than one HeLa infectious particle/cell, new virus first appears in the cell fraction 12 hr. after infection, and 4 hr. later virus is released into the medium. Virus in the cell fraction can be detected either by disruption of the cells, or simply by removing the cells from the glass with ethylenediaminetetra-acetic acid (EDTA) or saline. This ‘EDTA fraction’ may contain virus released from the cell surface. An attempt was made to determine the number of virus-yielding cells, by inoculation of whole cells on to the chick chorioallantoic membrane. The proportion which yielded virus was lower than would be expected from the input of HeLa-infectious virus. After removal of superficially attached virus with antiserum, it was not possible to detect infective virus during the latent period in the majority of cells which ultimately released virus.

Herpes virus is well known as a cause of latent infection, and it is our interest in latency which has prompted further investigation of the growth of this virus. Previous work on herpes virus multiplication has been carried out in the chorioallantoic membrane cavity of fertile hens’ eggs (Scott, Coriell, Blank & Gray, 1953; Wildy, 1954; Yoshino & Taniguchi, 1956a, b) and in explants of rabbit cornea (Scott, Burgoon, Coriell & Blank, 1953). In recent years a more quantitative approach has been possible through the use of cell suspensions and monolayer cultures, and growth studies have been reported by Gostling & Bedson (1956) and Gostling (1956) in chick embryo cells, and more recently by Kaplan (1957) in rabbit kidney cells. We have studied the growth of herpes virus in HeLa cells because of the desirability of propagating cells indefinitely when studying latent infection. HeLa cells are also convenient because they are suitable for isolation of cell clones, and for study of virus release from isolated single cells.

This paper describes some aspects of virus growth in monolayer cultures of HeLa cells. Studies on plaque formation in HeLa cells (Farnham, 1958), cytological observations (Ross & Orlans, 1958), chemical changes (Newton & Stoker, 1958), electron microscope studies (Stoker, Smith & Ross, 1958) and single cell experiments (Wildy, P., Stoker, M. G. P. & Ross, R. W., unpublished) will be given in other communications.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-19-2-250
1958-10-01
2024-11-11
Loading full text...

Full text loading...

/deliver/fulltext/micro/19/2/mic-19-2-250.html?itemId=/content/journal/micro/10.1099/00221287-19-2-250&mimeType=html&fmt=ahah

References

  1. Dulbecco R., Vogt M. 1954; Plaque formation and isolation of pure lines with poliomyelitis viruses. J. exp. Med 99:167
    [Google Scholar]
  2. Farnham A. E. 1958; The formation of microscopic plaques by herpes simplex virus in HeLa cells. Virology In the Press
    [Google Scholar]
  3. Gey G. O. 1949 Bull. Tissue Culture Ass16March Parker R. C. quoted by in Methods of Tissue Culture , 2nd ed. 1950, p. 81 New York: P. B. Hoeber, Inc;
    [Google Scholar]
  4. Gostling J. V. T. 1956; Intracellular site of developing herpes virus. Nature; Land: 1781238
    [Google Scholar]
  5. Gostling J. V. T., Bedson S. P. 1956; Observations on the mode of multiplication of herpes virus. Brit. J. exp. Path 37:434
    [Google Scholar]
  6. Kaplan A. S. 1957; A study of the herpes simplex virus-rabbit kidney cell system by the plaque technique. Virology 4:435
    [Google Scholar]
  7. Nadaje T., Tamm I., Overman J. R. 1955; A new technique for dropping the chorioallantoic membrane in embryonated chicken eggs. J. Lab. clin. Med 46:648
    [Google Scholar]
  8. Newton A., Stoker M. G. P. 1958; Changes in nucleic acid content of HeLa cells infected with herpes virus. Virology 5:549
    [Google Scholar]
  9. Ross R. W., Orlans E. 1958; The redistribution of nucleic acid and the appearance of specific antigen in HeLa cells infected with herpes virus. J. Path. Bact 76: in the press
    [Google Scholar]
  10. Scott T. F. McN., Burgoon C. F., Coriell L. L., Blank H. 1953; The growth curve of the virus of herpes simplex in rabbit comeal cells grown in tissue culture with parallel observations on the development of the intranuclear inclusion body. J. Immunol 71:385
    [Google Scholar]
  11. Scott T. F. McN., Coriell L. L., Blank H., Gray A. 1953; The growth curve of the virus of herpes simplex on the chorioallantoic membrane of the embryonated hen’s egg. J. Immunol 71:134
    [Google Scholar]
  12. Stoker M. G. P., Smith K., Ross R. W. 1958; Electron microscope studies of HeLa cells infected with herpes vims. J. gen. Microbiol 19:244
    [Google Scholar]
  13. Wildy P. 1954; The growth of herpes simplex vims. Aust. J. exp. Biol. med. Sci 32:605
    [Google Scholar]
  14. Yoshino K., Taniguchi H. 1956a; Quantitative studies on the interactions between herpes simplex vims and ectodermal cell of chorioallantoic membrane of fertile hen’s egg by means of the cover slip infection system. I. Adsorption of vims on to cells. Jap. J. med. Sci. Biol 9:303
    [Google Scholar]
  15. Yoshino K., Taniguchi H. 1956b; Quantitative studies on the interactions between herpes simplex virus and ectodermal cell of chorioallantoic membrane of fertile hen’s egg by means of the cover slip infection system. II. Growth curve with special reference to the demonstration of non-infectious phase by removal of surface vims. Jap. J. med. Sd. Biol 9:321
    [Google Scholar]
/content/journal/micro/10.1099/00221287-19-2-250
Loading
/content/journal/micro/10.1099/00221287-19-2-250
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