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. 1995 Mar;69(3):1762–1768. doi: 10.1128/jvi.69.3.1762-1768.1995

Latency, without persistence, of murine cytomegalovirus in the spleen and kidney.

J L Pollock 1, H W Virgin 4th 1
PMCID: PMC188782  PMID: 7853515

Abstract

It is not known if murine cytomegalovirus (MCMV) establishes a state of molecular latency independent of low-level persistent infection. The presence of low levels of infectious MCMV distinguishes persistence from molecular latency. Thus, the distinction between persistence and latency has depended on the sensitivity of plaque assays for detecting low levels of infectious virus in tissue of previously infected mice. To determine whether MCMV establishes molecular latency or remains persistent, we developed two assays for detecting low levels of MCMV in tissue. Using prolonged in vitro culture of virus with either mouse embryonic fibroblasts or the murine 3T12 fibroblast cell line, we reproducibly detected a single PFU of MCMV. Inclusion of undiluted sonicated tissue in this assay decreased sensitivity by up to 100-fold. However, sensitivity was improved to 1 PFU of MCMV when sonicated tissue was appropriately diluted. Severe combined immunodeficient (SCID) mice were also used to detect MCMV in sonicated tissue. Infection of SCID mice with a single PFU of MCMV killed two of eight SCID mice, and the 50% lethal dose of MCMV in SCID mice was 2 to 3 PFU. Applying these two methods, we detected infectious virus in 0 of 34 spleens, 1 of 34 kidneys, and 0 of 37 salivary glands from latently infected mice. Spleens and kidneys assessed for persistent virus contained MCMV DNA by PCR and reactivated after 10 to 50 days in explant cultures. Latently infected kidney cells reactivated after adoptive transfer to SCID mice. Quantitation of the MCMV genome by PCR showed that latently infected spleens without detectable infectious MCMV contained about 3,000,000 copies of the MCMV genome. These results demonstrate that MCMV latency in spleen and kidney exists in the absence of low-level persistent infection. Use of assays with defined sensitivity for detection of MCMV in tissue provides a basis for evaluation of cytomegalovirus gene expression in the spleen and kidney during molecular latency.

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Selected References

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  1. Balthesen M., Messerle M., Reddehase M. J. Lungs are a major organ site of cytomegalovirus latency and recurrence. J Virol. 1993 Sep;67(9):5360–5366. doi: 10.1128/jvi.67.9.5360-5366.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Custer R. P., Bosma G. C., Bosma M. J. Severe combined immunodeficiency (SCID) in the mouse. Pathology, reconstitution, neoplasms. Am J Pathol. 1985 Sep;120(3):464–477. [PMC free article] [PubMed] [Google Scholar]
  3. Heise M. T., Virgin H. W., 4th The T-cell-independent role of gamma interferon and tumor necrosis factor alpha in macrophage activation during murine cytomegalovirus and herpes simplex virus infections. J Virol. 1995 Feb;69(2):904–909. doi: 10.1128/jvi.69.2.904-909.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Henry S. C., Hamilton J. D. Detection of murine cytomegalovirus immediate early 1 transcripts in the spleens of latently infected mice. J Infect Dis. 1993 Apr;167(4):950–954. doi: 10.1093/infdis/167.4.950. [DOI] [PubMed] [Google Scholar]
  5. Jonjić S., Mutter W., Weiland F., Reddehase M. J., Koszinowski U. H. Site-restricted persistent cytomegalovirus infection after selective long-term depletion of CD4+ T lymphocytes. J Exp Med. 1989 Apr 1;169(4):1199–1212. doi: 10.1084/jem.169.4.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jonjić S., Pavić I., Polić B., Crnković I., Lucin P., Koszinowski U. H. Antibodies are not essential for the resolution of primary cytomegalovirus infection but limit dissemination of recurrent virus. J Exp Med. 1994 May 1;179(5):1713–1717. doi: 10.1084/jem.179.5.1713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jordan M. C. Adverse effects of cytomegalovirus vaccination in mice. J Clin Invest. 1980 Apr;65(4):798–803. doi: 10.1172/JCI109730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jordan M. C. Latent infection and the elusive cytomegalovirus. Rev Infect Dis. 1983 Mar-Apr;5(2):205–215. doi: 10.1093/clinids/5.2.205. [DOI] [PubMed] [Google Scholar]
  9. Jordan M. C., Mar V. L. Spontaneous activation of latent cytomegalovirus from murine spleen explants. Role of lymphocytes and macrophages in release and replication of virus. J Clin Invest. 1982 Oct;70(4):762–768. doi: 10.1172/JCI110672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jordan M. C., Shanley J. D., Stevens J. G. Immunosuppression reactivates and disseminates latent murine cytomegalovirus. J Gen Virol. 1977 Nov;37(2):419–423. doi: 10.1099/0022-1317-37-2-419. [DOI] [PubMed] [Google Scholar]
  11. Keil G. M., Ebeling-Keil A., Koszinowski U. H. Sequence and structural organization of murine cytomegalovirus immediate-early gene 1. J Virol. 1987 Jun;61(6):1901–1908. doi: 10.1128/jvi.61.6.1901-1908.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klotman M. E., Henry S. C., Greene R. C., Brazy P. C., Klotman P. E., Hamilton J. D. Detection of mouse cytomegalovirus nucleic acid in latently infected mice by in vitro enzymatic amplification. J Infect Dis. 1990 Feb;161(2):220–225. doi: 10.1093/infdis/161.2.220. [DOI] [PubMed] [Google Scholar]
  13. Koszinowski U. H., Keil G. M., Volkmer H., Fibi M. R., Ebeling-Keil A., Münch K. The 89,000-Mr murine cytomegalovirus immediate-early protein activates gene transcription. J Virol. 1986 Apr;58(1):59–66. doi: 10.1128/jvi.58.1.59-66.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mayo D., Armstrong J. A., Ho M. Activation of latent murine cytomegalovirus infection: cocultivation, cell transfer, and the effect of immunosuppression. J Infect Dis. 1978 Dec;138(6):890–896. doi: 10.1093/infdis/138.6.890. [DOI] [PubMed] [Google Scholar]
  15. Mercer J. A., Wiley C. A., Spector D. H. Pathogenesis of murine cytomegalovirus infection: identification of infected cells in the spleen during acute and latent infections. J Virol. 1988 Mar;62(3):987–997. doi: 10.1128/jvi.62.3.987-997.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pomeroy C., Hilleren P. J., Jordan M. C. Latent murine cytomegalovirus DNA in splenic stromal cells of mice. J Virol. 1991 Jun;65(6):3330–3334. doi: 10.1128/jvi.65.6.3330-3334.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rubin R. H. Impact of cytomegalovirus infection on organ transplant recipients. Rev Infect Dis. 1990 Sep-Oct;12 (Suppl 7):S754–S766. doi: 10.1093/clinids/12.supplement_7.s754. [DOI] [PubMed] [Google Scholar]
  18. SMITH M. G. Propagation of salivary gland virus of the mouse in tissue cultures. Proc Soc Exp Biol Med. 1954 Jul;86(3):435–440. doi: 10.3181/00379727-86-21123. [DOI] [PubMed] [Google Scholar]
  19. Schooley R. T. Cytomegalovirus in the setting of infection with human immunodeficiency virus. Rev Infect Dis. 1990 Sep-Oct;12 (Suppl 7):S811–S819. doi: 10.1093/clinids/12.supplement_7.s811. [DOI] [PubMed] [Google Scholar]
  20. Shanley J. D., Jordan M. C., Cook M. L., Stevens J. G. Pathogenesis of reactivated latent murine cytomegalovirus infection. Am J Pathol. 1979 Apr;95(1):67–80. [PMC free article] [PubMed] [Google Scholar]
  21. Tokunaga K., Taniguchi H., Yoda K., Shimizu M., Sakiyama S. Nucleotide sequence of a full-length cDNA for mouse cytoskeletal beta-actin mRNA. Nucleic Acids Res. 1986 Mar 25;14(6):2829–2829. doi: 10.1093/nar/14.6.2829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Vieira J., Farrell H. E., Rawlinson W. D., Mocarski E. S. Genes in the HindIII J fragment of the murine cytomegalovirus genome are dispensable for growth in cultured cells: insertion mutagenesis with a lacZ/gpt cassette. J Virol. 1994 Aug;68(8):4837–4846. doi: 10.1128/jvi.68.8.4837-4846.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wise T. G., Manischewitz J. E., Quinnan G. V., Aulakh G. S., Ennis F. A. Latent cytomegalovirus infection of BALB/c mouse spleens detected by an explant culture technique. J Gen Virol. 1979 Aug;44(2):551–556. doi: 10.1099/0022-1317-44-2-551. [DOI] [PubMed] [Google Scholar]
  24. Yuhasz S. A., Dissette V. B., Cook M. L., Stevens J. G. Murine cytomegalovirus is present in both chronic active and latent states in persistently infected mice. Virology. 1994 Jul;202(1):272–280. doi: 10.1006/viro.1994.1343. [DOI] [PubMed] [Google Scholar]

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