Summary
Three human hybrid cell lines were generated by the fusion of D 98OR, a HeLa cell variant, and TIG human diploid fibroblasts. Chromosome numbers of the hybrid cells fell between that of D 98OR cells and the combined chromosome number of the two cell lines, with three marker chromosomes identical to those of D 98OR cells. One hybrid, B-3, produced large amounts of human fibronectin as analyzed by immunohistochemistry. Only human cytomegalovirus (HCMV) infected B-3 cells showed positive fluorescence as detected by human antiserum to HCMV. Further cloning of B-3 by limiting dilution resulted in two cloned hybrids with markedly enhanced virus production as compared with B-3 cells. Treatment of these two clones with phorbol ester further enhanced virus production. These cloned hybrids may provide a tool to analyze host cell factors controlling the transcription and replication of HCMV.
Similar content being viewed by others
References
Amtmann E, Sauer G (1982) Activation of non-expressed bovine papilloma virus genomes by tumour promoters. Nature 296: 675–677
Epstein AL, Jacquement B, Patet J (1985) Susceptibility to herpes simplex virus type 1 infection of non-permissive rat XC (HPRT−) X permissive mouse L (TK−) hybrid cells. J Gen Virol 66: 1805–1809
Fisher PB, Weinstein IB, Eisenberg D, Ginsberg HS (1978) Interactions between adenovirus, a tumor promoter, and chemical carcinogens in transformation of rat embryo cell cultures. Proc Natl Acad Sci USA 75: 2311–2314
Gönczöl E, Andrews PW, Plotkin SA (1984) Cytomegalovirus replicates in differentiated but not undifferentiated human embryonal carcinoma cells. Science 224: 159–161
Hynes R, Yamada K (1982) Fibronectin: multifunctional modular glycoproteins. J Cell Biol 95: 369–377
Ihara S, Saito S, Watanabe Y (1982) Suppression of fibronectin synthesis by an early function(s) of human cytomegalovirus. J Gen Virol 59: 409–413
Imbra RJ, Karin M (1986) Phorbol ester induces the transcriptional stimulatory activity of the SV 40 enhancer. Nature 323: 555–558
Kaji K, Mitsuyoshi M (1980) A low-density inoculation method for the serial subcultivation of human diploid fibroblasts: an efficient model system for the study of culture ageing. Mech Ageing Dev 13: 219–225
Kimata K, Sakakura T, Inaguma Y, Kato M, Nishizuka Y (1985) Participation of two different mesenchymes in the developing mouse mammary glands: synthesis of basement menbrane components by fat pad precursor cells. J Embryol Exp Morphol 89: 243–257
Knowlers WA (1976)In vitro cultivation of human cytomegalovirus in thyroid epithelial cells. Arch Virol 50: 119–124
Law LF, Nathans D (1985) Identification of a set of genes expressed during the G0-G1 transition of cultured mouse embryo. EMBO J 4: 3145–3151
Lin JC, Show JE, Smith MC, Pagano JS (1979) Effect of 12-O-tetradecanoyl-phorbol-13-acetate on the replication of Epstein-Barr virus. I. Characterization of viral DNA. Virology 99: 183–187
Lin JC, Smith MC, Pagano JS (1982) Effect of 12-O-tetradecanoyl-phorbol-13-acetate on cell proliferation and Epstein-Barr virus DNA replication. Virology 117: 186–194
Michelson-Fiske S, Arnault J, Febvre H (1975) Cytomegalovirus infection of human lung epithelial cellsin vitro. Intervirology 5: 354–363
Nakane PK (1975) Recent progress in the peroxidase-labeled antibody method. Ann NY Acad Sci 254: 203–211
Nishizuka Y (1984) The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature 308: 693–698
Smith MG (1959) The salivary gland viruses of man and animals (cytomegalic inclusion disease). Prog Med Virol 2: 171–202
St Jeor S, Rapp F (1973) Cytomegalovirus replication in cells pretreated with 5-iodo-2′-deoxyuridine. J Virol 11: 986–990
St Jeor S, Rapp F (1973) Cytomegalovirus: conversion of nonpermissive cells to a permissive state for virus replication. Science 181: 1060–1061
Stanbridge EJ, Der CJ, Doersen CJ, Nishimi RY, Peehl DM, Weissman BE, Wilkinson JE (1982) Human cell hybrids: analysis of transformation and tumorigenicity. Science 215: 252–259
Vonka V, Anisimova E, Mackek M (1976) Replication of cytomegalovirus in a human epithelioid diploid cell line. Arch Virol 52: 283–296
Weissman B, Stanbridge EJ (1980) Characterization of ouabain-resistant, hypoxanthinephosphorybosyltransferase-deficient human cells and their usefulness as a general method for production of human cell hybrids. Cytogenet Cell Genet 28: 227–239
Weller TH (1971) The cytomegalovirus: ubiquitous agents with protean clinical manifestations. N Engl J Med 285: 203–214
Wentworth BB, French L (1970) Plaque assay of cytomegalovirus strains of human origin. Proc Soc Exp Biol Med 135: 253–258
Zur Hausen H, O'Neill FJ, Freese UK, Hecker E (1978) Persisting oncogenic herpes-virus induced by tumor promoter TPA. Nature 272: 373–375
Zur Hausen H, Yamamoto N, Bauer G (1982) Virus induction by tumor promoter. Carcinogenesis 7: 617–623
Author information
Authors and Affiliations
Additional information
With 5 Figures
Rights and permissions
About this article
Cite this article
Tsutsui, Y., Sonta, S., Kashiwai, A. et al. Viral replication in HeLa/fibroblast hybrid cells infected with human cytomegalovirus. Archives of Virology 95, 29–40 (1987). https://doi.org/10.1007/BF01311332
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01311332