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. 1988 Dec;26(12):2510–2514. doi: 10.1128/jcm.26.12.2510-2514.1988

Enhanced detection of cytomegalovirus in confluent MRC-5 cells treated with dexamethasone and dimethyl sulfoxide.

P G West 1, B Aldrich 1, R Hartwig 1, G J Haller 1
PMCID: PMC266935  PMID: 2852669

Abstract

Optimum growth conditions for human cytomegalovirus (HCMV) include the use of subconfluent, actively growing cultures of human embryonic fibroblasts. Many clinical virology laboratories, however, use tissue culture cells from commercial sources. These cells are usually confluent, static cultures that tend to be less sensitive to viral infection. To determine whether dimethyl sulfoxide (DMSO) or dexamethasone (DEX), which are known enhancers of HCMV, facilitates the detection of the virus in confluent cells, we tested both HCMV AD169 and a number of clinical specimens suspected to contain HCMV on MRC-5 cells in both shell vials and conventional tube cultures. We found that, in the shell vial test, treatment of the cultures with either DMSO or DEX before and after inoculation increased the number of cells staining positive by three- to sixfold compared with untreated controls. Best results were obtained by pretreating the cultures with DEX alone and by treating the cultures with a combination of DEX and 1% DMSO postinfection. In the conventional MRC-5 culture tubes, treatment with the reagents resulted in the more rapid appearance of cytopathic effect and a more extensive infection of the cell sheet. The experimental findings indicate that the enhancing effect of DEX is attributable mainly to the increased production of a cellular mRNA during the period preceding viral infection.

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Selected References

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  1. Abelson H. T., Penman S. Messenger RNA formation: resistance to inhibition by 3'-deoxycytidine. Biochim Biophys Acta. 1972 Aug 16;277(1):129–133. doi: 10.1016/0005-2787(72)90359-0. [DOI] [PubMed] [Google Scholar]
  2. DeMarchi J. M., Kaplan A. S. Physiological state of human embryonic lung cells affects their response to human cytomegalovirus. J Virol. 1977 Jul;23(1):126–132. doi: 10.1128/jvi.23.1.126-132.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Furukawa T., Tanaka S., Plotkin S. A. Resticted growth of human cytomegalovirus in UV-irradiated WI-38 human fibroblasts. Proc Soc Exp Biol Med. 1975 Apr;148(4):1249–1251. doi: 10.3181/00379727-148-38726. [DOI] [PubMed] [Google Scholar]
  4. Gleaves C. A., Smith T. F., Shuster E. A., Pearson G. R. Rapid detection of cytomegalovirus in MRC-5 cells inoculated with urine specimens by using low-speed centrifugation and monoclonal antibody to an early antigen. J Clin Microbiol. 1984 Jun;19(6):917–919. doi: 10.1128/jcm.19.6.917-919.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Higgins S. J., Gehring U. Molecular mechanisms of steroid hormone action. Adv Cancer Res. 1978;28:313–397. doi: 10.1016/s0065-230x(08)60650-8. [DOI] [PubMed] [Google Scholar]
  6. Jeang K. T., Rawlins D. R., Rosenfeld P. J., Shero J. H., Kelly T. J., Hayward G. S. Multiple tandemly repeated binding sites for cellular nuclear factor 1 that surround the major immediate-early promoters of simian and human cytomegalovirus. J Virol. 1987 May;61(5):1559–1570. doi: 10.1128/jvi.61.5.1559-1570.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Parks W. P., Ransom J. C., Young H. A., Scolnick E. M. Mammary tumor virus induction by glucocorticoids. Characterization of specific transcriptional regulation. J Biol Chem. 1975 May 10;250(9):3330–3336. [PubMed] [Google Scholar]
  8. Parks W. P., Scolnick E. M., Kozikowski E. H. Dexamethasone stimulation of murine mammary tumor virus expression: a tissue culture source of virus. Science. 1974 Apr 12;184(4133):158–160. doi: 10.1126/science.184.4133.158. [DOI] [PubMed] [Google Scholar]
  9. Scott W. A., Shields R., Tomkins G. M. Mechanism of hormonal induction of tyrosine aminotransferase studied by measurement of the concentration of growing enzyme molecules. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2937–2941. doi: 10.1073/pnas.69.10.2937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Tanaka J., Kamiya S., Ogura T., Sato H., Ogura H., Hatano M. Effect of dimethyl sulfoxide on interaction of human cytomegalovirus with host cell: conversion of a nonproductive state of cell to a productive state for virus replication. Virology. 1985 Oct 30;146(2):165–176. doi: 10.1016/0042-6822(85)90001-7. [DOI] [PubMed] [Google Scholar]
  11. Tanaka J., Ogura T., Kamiya S., Sato H., Yoshie T., Ogura H., Hatano M. Enhanced replication of human cytomegalovirus in human fibroblasts treated with dexamethasone. J Gen Virol. 1984 Oct;65(Pt 10):1759–1767. doi: 10.1099/0022-1317-65-10-1759. [DOI] [PubMed] [Google Scholar]
  12. Tanaka J., Ogura T., Kamiya S., Yoshie T., Yabuki Y., Hatano M. Dexamethasone enhances human cytomegalovirus replication in human epithelial cell cultures. Virology. 1984 Jul 30;136(2):448–452. doi: 10.1016/0042-6822(84)90182-x. [DOI] [PubMed] [Google Scholar]

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