1887

Abstract

SUMMARY

Human cytomegalovirus (HCMV) infection has previously been associated with the production of immunosuppression. The mechanism by which any such immuno-suppressive effect might be mediated is unclear but previous work has implicated an effect of the virus on monocytes. We have attempted to characterize the immunosuppressive activity produced by infection of normal monocytes with HCMV strain AD169. We first examined the ability of HCMV AD169 and recent clinical isolates to infect normal peripheral blood mononuclear cells . We have found by immunofluorescence analysis that only a very limited number of peripheral blood mononuclear cells (0·2 to 0·5%) showed evidence of virus infection as demonstrated by expression of the major immediate early protein. We found that the inhibitory activity of supernatants of monocytes exposed to HCMV which suppressed mitogen-driven T cell responsiveness was associated with a protein of about 95K. Experiments to investigate the mechanism of action of this inhibitor suggested the possibility of mycoplasma contamination and we were subsequently able to isolate from our AD169 virus stock. When a series of low passage clinical isolates of HCMV were examined for their ability to cause immunosuppression, there was a direct correlation between suppression and the presence of contaminating mycoplasmas. Using mycoplasma-free isolates of HCMV we could demonstrate no immunosuppressive effect on mitogen-mediated T cell proliferation of both unseparated human peripheral blood lymphocytes and nylon wool non-adherent T cells; these virus isolates also did not suppress accessory cell function or interleukin 1 production by monocytes infected . We conclude that the previously reported immunosuppressive effects of HCMV may be attributable to the presence of mycoplasmas and are unlikely to be due to expression of HCMV in monocytes. We suggest that mycoplasma contamination of isolates of HCMV may be a more extensive problem than is currently recognized.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-70-3-685
1989-03-01
2024-12-06
Loading full text...

Full text loading...

/deliver/fulltext/jgv/70/3/JV0700030685.html?itemId=/content/journal/jgv/10.1099/0022-1317-70-3-685&mimeType=html&fmt=ahah

References

  1. Amento E. P., Kurnick J. T., Krane S. M. 1985; Interleukin 1 production by the human monocyte line U937 requires a lymphokine induction signal distinct from interleukin 2 or interferons. Journal of Immunology 134:350–357
    [Google Scholar]
  2. Barile M. F. 1981; Mycoplasma infections of cell cultures. Israel Journal of Medical Science 17:555
    [Google Scholar]
  3. Barile M. F., Leventhal B. G. 1968; Possible mechanism for mycoplasma inhibition of lymphocyte transformation induced by phytohaemagglutinin. Nature, London 219:751–752
    [Google Scholar]
  4. Boyum A. 1973; Separation of blood leukocytes, granulocytes and lymphocytes. Tissue Antigens 4:269–274
    [Google Scholar]
  5. Carney W. P., Hirsch M. S. 1981; Mechanisms of immunosuppression in CMV mononucleosis. II. Virus- monocyte interactions. Journal of Infectious Diseases 144:47–54
    [Google Scholar]
  6. Chen T. R. 1977; In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. Experimental Cell Research 14:255–262
    [Google Scholar]
  7. Einhorn L., Ost A. 1984; Cytomegalovirus infection of human blood cells. Journal of Infectious Diseases 149:207–214
    [Google Scholar]
  8. Falkoff R. J. M., Peters M., Fauci A. S. 1982; T cell enrichment and depletion of human peripheral blood mononuclear cell preparations. Unexpected findings in the study of the functional properties of the separated population. Journal of Immunological Methods 50:39–49
    [Google Scholar]
  9. Friis N. F. 1975; Some recommendations concerning primary isolation of Mycoplasma suipneumoniae and Mycoplasma flocculare. Nordisk veterinaermedicin 27:337–339
    [Google Scholar]
  10. Gourlay R. N., Leach R. H., Howard C. J. 1974; Mycoplasma verecundum, a new species isolated from bovine eyes. Journal of General Microbiology 81:475–484
    [Google Scholar]
  11. Ho M. 1984; Immunology of cytomegalovirus. Birth Defects 20:131–147
    [Google Scholar]
  12. Howard R. J., Najarian I. S. 1974; Cytomegalovirus induced immunosuppression. 1. Humoral immunity. Clinical and Experimental Immunology 18:109–118
    [Google Scholar]
  13. Howard R. J., Miller J., Najarian J. S. 1974; Cytomegalovirus-induced immune suppression. II. Cell mediated immunity. Clinical and Experimental Immunology 18:119–126
    [Google Scholar]
  14. Julius M. H., Simpson E., Herzenberg L. A. 1973; A rapid method for the isolation of functional thymus- derived lymphocytes. European Journal of Immunology 3:645–649
    [Google Scholar]
  15. Koren H. S., Anderson S. J., Larrick J. W. 1979; In vitro activation of a macrophage-like cell line. Nature, London 279:328–331
    [Google Scholar]
  16. Rice G. P. A., Schrier R. D., Oldstone M. B. A. 1984; CMV infection of human lymphocytes and monocytes: virus expression is restricted to immediate early gene products. Proceedings of the National Academy of SciencesU.S.A 816134–6138
    [Google Scholar]
  17. Rinaldo C. R. Jr, Black P. H., Hirsch M. S. 1977; Interaction of cytomegalovirus with leukocytes from patients with mononucleosis due to cytomegalovirus. Journal of Infectious Diseases 136:667–678
    [Google Scholar]
  18. Rodgers B. C., Scott D. M., Mundin J., Sissons J. G. P. 1985; Monocyte-derived inhibitor of interleukin 1 induced by human cytomegalovirus. Journal of Virology 55:527–532
    [Google Scholar]
  19. Scala G., Kuang Y. D., Hall R. E., Muchmore A. V., Oppenheim J. J. 1984; Accessory cell function of human B cells. 1. Production of both interleukin 1-like activity and an interleukin 1 inhibitory factor by an EBV- transformed human B cell line. Journal of Experimental Medicine 159:1637–1652
    [Google Scholar]
  20. Schooley R. T. 1983; Association of herpesvirus infections with T lymphocyte subset alterations, glomerulonephropathy and opportunistic infections after renal transplantation. New England Journal of Medicine 308:307–313
    [Google Scholar]
  21. Schrier R. D., Oldstone M. B. A. 1986; Recent clinical isolates of cytomegalovirus suppress human cytomegalovirus-specific human leukocyte antigen-restricted cytotoxic T-lymphocyte activity. Journal of Virology 59:127–131
    [Google Scholar]
  22. Schrier R. D., Nelson J. A., Oldstone M. B. A. 1985; Detection of human cytomegalovirus in peripheral blood lymphocytes in a natural infection. Science 230:1048–1051
    [Google Scholar]
  23. Schrier R. D., Rice G. P. A., Oldstone M. B. A. 1986; Suppression of natural killer cell activity and T cell proliferation by fresh isolates of human cytomegalovirus. Journal of Infectious Diseases 153:1084–1091
    [Google Scholar]
  24. Sinigaglia F., Talmadge K. W. 1985; Inhibition of 3H thymidine incorporation by Mycoplasma arginini-infected cells due to enzymatic cleavage of the nucleoside. European Journal of Immunology 15:692–696
    [Google Scholar]
  25. Smith P. D., Miyake Y., Rook A. H., Armstrong G. R., Manischewitz J. F., Liu J. R., Pearson G., Wahl L. M., Wahl S. M. 1985; Cytomegalovirus infection depresses monocyte production of interleukin 1. In Progress in Leucocyte Biology 2 The Physiologic, Metabolic and Immunologic Actions of Interleukin 1319–326 Kluger M. J., Oppenheim J. J., Powanda M. C. New York: Alan R. Liss;
    [Google Scholar]
  26. Starr S. F., Arenzana-Seisedos F., Michelson S., Virelizier J. L. 1985; Effect of human cytomegalovirus on monocyte interleukin 1 secretion. In Progress in Leucocyte Biology 2 The Physiologic, Metabolic and Immunologic Actions of Interleukin 1327–338 Kluger M. J., Oppenheim J. J., Powanda M. C. Yew York: Alan R. Less;
    [Google Scholar]
  27. Ward P. 1981; The influence of fixation and composition of incubation medium on α-naphthyl acetate esterase staining of T-lymphocytes and monocytes in human peripheral blood. Journal of Immunological Methods 41:23–27
    [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-70-3-685
Loading
/content/journal/jgv/10.1099/0022-1317-70-3-685
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