Abstract
Immunocompromised patients with disseminated human cytomegalovirus (HCMV) infection have circulating PMN carrying HCMV pp65 (antigenemia), infectious virus (viremia), and viral DNA (leukoDNAemia). Because HCMV does not fully replicate in PMN, it is generally hypothesized that virions and viral materials are taken up by phagocytosis from fully permissive HCMV-infected endothelial cells. However, no experimental evidence has ever been provided for these PMN-endothelium interactions. PMN from 11 donors were cocultured with endothelial cells infected with an endothelium-adapted HCMV strain and with human fibroblasts infected with low-passaged clinical and laboratory-adapted HCMV strains. pp65-positive PMN were detected after coculture with both HCMV-infected endothelial and fibroblast cells, provided that wild and not laboratory-adapted strains were used. In addition, cocultured PMN carried infectious virus as demonstrated by virus isolation and presence of complete virus particles by electron microscopy. Moreover, high levels of viral DNA were consistently detected by quantitative PCR in cocultured PMN. Thus, we have generated in vitro the three most important viral parameters detected in patients with disseminated HCMV infection (antigenemia, viremia, and leukoDNAemia). The failure of laboratory-adapted HCMV strain to induce this phenomenon demonstrates that important modifications have occurred in attenuated viral strains affecting basic biological functions.
Full Text
The Full Text of this article is available as a PDF (335.4 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Brown J. M., Kaneshima H., Mocarski E. S. Dramatic interstrain differences in the replication of human cytomegalovirus in SCID-hu mice. J Infect Dis. 1995 Jun;171(6):1599–1603. doi: 10.1093/infdis/171.6.1599. [DOI] [PubMed] [Google Scholar]
- Cha T. A., Tom E., Kemble G. W., Duke G. M., Mocarski E. S., Spaete R. R. Human cytomegalovirus clinical isolates carry at least 19 genes not found in laboratory strains. J Virol. 1996 Jan;70(1):78–83. doi: 10.1128/jvi.70.1.78-83.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen T. R. In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. Exp Cell Res. 1977 Feb;104(2):255–262. doi: 10.1016/0014-4827(77)90089-1. [DOI] [PubMed] [Google Scholar]
- Elek S. D., Stern H. Development of a vaccine against mental retardation caused by cytomegalovirus infection in utero. Lancet. 1974 Jan 5;1(7845):1–5. doi: 10.1016/s0140-6736(74)92997-3. [DOI] [PubMed] [Google Scholar]
- Friedman H. M., Macarak E. J., MacGregor R. R., Wolfe J., Kefalides N. A. Virus infection of endothelial cells. J Infect Dis. 1981 Feb;143(2):266–273. doi: 10.1093/infdis/143.2.266. [DOI] [PubMed] [Google Scholar]
- Gerna G., Baldanti F., Sarasini A., Furione M., Percivalle E., Revello M. G., Zipeto D., Zella D. Effect of foscarnet induction treatment on quantitation of human cytomegalovirus (HCMV) DNA in peripheral blood polymorphonuclear leukocytes and aqueous humor of AIDS patients with HCMV retinitis. The Italian Foscarnet Study Group. Antimicrob Agents Chemother. 1994 Jan;38(1):38–44. doi: 10.1128/aac.38.1.38. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerna G., Furione M., Baldanti F., Sarasini A. Comparative quantitation of human cytomegalovirus DNA in blood leukocytes and plasma of transplant and AIDS patients. J Clin Microbiol. 1994 Nov;32(11):2709–2717. doi: 10.1128/jcm.32.11.2709-2717.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerna G., Revello M. G., Palla M., Percivalle E., Torsellini M. Microneutralization as a reference method for selection of the cut-off of an enzyme-linked immunosorbent assay for detection of IgG antibody to human cytomegalovirus. Microbiologica. 1992 Apr;15(2):177–181. [PubMed] [Google Scholar]
- Gerna G., Revello M. G., Percivalle E., Morini F. Comparison of different immunostaining techniques and monoclonal antibodies to the lower matrix phosphoprotein (pp65) for optimal quantitation of human cytomegalovirus antigenemia. J Clin Microbiol. 1992 May;30(5):1232–1237. doi: 10.1128/jcm.30.5.1232-1237.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerna G., Revello M. G., Percivalle E., Zavattoni M., Parea M., Battaglia M. Quantification of human cytomegalovirus viremia by using monoclonal antibodies to different viral proteins. J Clin Microbiol. 1990 Dec;28(12):2681–2688. doi: 10.1128/jcm.28.12.2681-2688.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerna G., Zipeto D., Percivalle E., Parea M., Revello M. G., Maccario R., Peri G., Milanesi G. Human cytomegalovirus infection of the major leukocyte subpopulations and evidence for initial viral replication in polymorphonuclear leukocytes from viremic patients. J Infect Dis. 1992 Dec;166(6):1236–1244. doi: 10.1093/infdis/166.6.1236. [DOI] [PubMed] [Google Scholar]
- Grefte A., van der Giessen M., van Son W., The T. H. Circulating cytomegalovirus (CMV)-infected endothelial cells in patients with an active CMV infection. J Infect Dis. 1993 Feb;167(2):270–277. doi: 10.1093/infdis/167.2.270. [DOI] [PubMed] [Google Scholar]
- Grundy J. E., Downes K. L. Up-regulation of LFA-3 and ICAM-1 on the surface of fibroblasts infected with cytomegalovirus. Immunology. 1993 Mar;78(3):405–412. [PMC free article] [PubMed] [Google Scholar]
- Percivalle E., Revello M. G., Vago L., Morini F., Gerna G. Circulating endothelial giant cells permissive for human cytomegalovirus (HCMV) are detected in disseminated HCMV infections with organ involvement. J Clin Invest. 1993 Aug;92(2):663–670. doi: 10.1172/JCI116635. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Plotkin S. A., Farquhar J., Horberger E. Clinical trials of immunization with the Towne 125 strain of human cytomegalovirus. J Infect Dis. 1976 Nov;134(5):470–475. doi: 10.1093/infdis/134.5.470. [DOI] [PubMed] [Google Scholar]
- Radsak K., Fuhrmann R., Franke R. P., Schneider D., Kollert A., Brücher K. H., Drenckhahn D. Induction by sodium butyrate of cytomegalovirus replication in human endothelial cells. Arch Virol. 1989;107(1-2):151–158. doi: 10.1007/BF01313887. [DOI] [PubMed] [Google Scholar]
- Revello M. G., Percivalle E., Di Matteo A., Morini F., Gerna G. Nuclear expression of the lower matrix protein of human cytomegalovirus in peripheral blood leukocytes of immunocompromised viraemic patients. J Gen Virol. 1992 Feb;73(Pt 2):437–442. doi: 10.1099/0022-1317-73-2-437. [DOI] [PubMed] [Google Scholar]
- Sedmak D. D., Knight D. A., Vook N. C., Waldman J. W. Divergent patterns of ELAM-1, ICAM-1, and VCAM-1 expression on cytomegalovirus-infected endothelial cells. Transplantation. 1994 Dec 27;58(12):1379–1385. [PubMed] [Google Scholar]
- Span A. H., Van Boven C. P., Bruggeman C. A. The effect of cytomegalovirus infection on the adherence of polymorphonuclear leucocytes to endothelial cells. Eur J Clin Invest. 1989 Dec;19(6):542–548. doi: 10.1111/j.1365-2362.1989.tb00272.x. [DOI] [PubMed] [Google Scholar]
- Span A. H., van Dam-Mieras M. C., Mullers W., Endert J., Muller A. D., Bruggeman C. A. The effect of virus infection on the adherence of leukocytes or platelets to endothelial cells. Eur J Clin Invest. 1991 Jun;21(3):331–338. doi: 10.1111/j.1365-2362.1991.tb01378.x. [DOI] [PubMed] [Google Scholar]
- Springer T. A. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994 Jan 28;76(2):301–314. doi: 10.1016/0092-8674(94)90337-9. [DOI] [PubMed] [Google Scholar]
- Turtinen L. W., Saltzman R., Jordan M. C., Haase A. T. Interactions of human cytomegalovirus with leukocytes in vivo: analysis by in situ hybridization. Microb Pathog. 1987 Oct;3(4):287–297. doi: 10.1016/0882-4010(87)90062-3. [DOI] [PubMed] [Google Scholar]
- Waldman W. J., Knight D. A., Huang E. H., Sedmak D. D. Bidirectional transmission of infectious cytomegalovirus between monocytes and vascular endothelial cells: an in vitro model. J Infect Dis. 1995 Feb;171(2):263–272. doi: 10.1093/infdis/171.2.263. [DOI] [PubMed] [Google Scholar]
- Waldman W. J., Roberts W. H., Davis D. H., Williams M. V., Sedmak D. D., Stephens R. E. Preservation of natural endothelial cytopathogenicity of cytomegalovirus by propagation in endothelial cells. Arch Virol. 1991;117(3-4):143–164. doi: 10.1007/BF01310761. [DOI] [PubMed] [Google Scholar]
- van der Bij W., Torensma R., van Son W. J., Anema J., Schirm J., Tegzess A. M., The T. H. Rapid immunodiagnosis of active cytomegalovirus infection by monoclonal antibody staining of blood leucocytes. J Med Virol. 1988 Jun;25(2):179–188. doi: 10.1002/jmv.1890250208. [DOI] [PubMed] [Google Scholar]