Summary
Cell culture systems were developed for rapid antiviral drug screening, using murine cytomegalovirus (MCMV) as an alternative to the slower growing human CMV. Since previous assay methods with MCMV employed mouse embryo fibroblasts (MEF cells), which are labor intensive to prepare and die off after 3–4 passages from primary culture, identification of virus-susceptible continuous cell lines was desirable. Three cell lines were found useful for assaying MCMV: C127I, SC-1, and 3T3. The antiviral agents acyclovir, ganciclovir, 5-fluoroarabinofuranosylcytosine, and 2′-fluoro-2′-deoxy-5-iodoarabinofuranosylcytosine were evaluated in the 3 continuous cell lines and in MEF cells. The 50% virus- or cell-inhibitory concentration values determined for each compound did not vary much from cell to cell. MEF cells were 10-fold more sensitive than the other cell lines to quantify virus from mouse organs, however. Virus propagated in 3T3 and SC-1 cells were as virulent to mice as salivary gland virus, whereas virus from MEF and C127I cells was more attenuated. Overall, C127I cells were judged to be the best for large scale antiviral screening in vitro, but MEF was the cell type of choice for titration of viruses from mouse organs and tissues.
Similar content being viewed by others
References
Cheng Y-C, Huang E-S, Lin J-C, Mar E-C, Pagano JS, Dutschman GE, Grill SP (1983) Unique spectrum of activity of 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine against herpesviruses in vitro and its mode of action against herpes simplex virus type 1. Proc Natl Acad Sci USA 80: 2767–2770
Colacino JM, Lopez C (1983) Efficacy and selectivity of some nucleoside analogs as anti-human cytomegalovirus agents. Antimicrob Agents Chemother 24: 505–508
Duke AE, Smee DF, Chernow M, Boehme R, Matthews TR (1986) In vitro and in vivo activities of phosphate derivatives of 9-(1,3-dihydroxy-2-propoxymethyl)guanine against cytomegaloviruses. Antiviral Res 6: 299–308
Freitas VR, Smee DF, Chernow M, Boehme R, Matthews TR (1985) Activity of 9-(1,3-dihydroxy-2-propoxymethyl)guanine compared with that of acyclovir against human, monkey and rodent cytomegaloviruses. Antimicrob Agents Chemother 28: 240–245
Hudson JB (1988) Further studies on the mechanism of centrifugal enhancement of cytomegalovirus infectivity. J Virol Methods 19: 97–108
Kelsey DK, Kern ER, Overall JC Jr, Glasgow LA (1976) Effect of cytosine arabinoside and 5-iodo-2′-deoxyuridine on a cytomegalovirus infection in newborn mice. Antimicrob Agents Chemother 9: 458–464
Laskin OL, Stahl-Bayliss CM, Kalman CM, Rosecan LR (1987) Use of ganciclovir to treat serious cytomegalovirus infections in patients with AIDS. J Infect Dis 155: 323–327
Macher AM, Reichert CM, Straus SE, Longo DL, Parillo J, Lane HC, Fauci AS, Rook AH, Manischewitz J, Quinnan GV (1983) Death in the AIDS patient: role of cytomegalovirus. N Engl J Med 309: 1454
Overall JC Jr, Kern ER, Glasgow LA (1976) Effective antiviral chemotherapy in cytomegalovirus infection in mice. J Infect Dis 133: A237–244
Raju N, Smee DF, Robins RK, Vaghefi MM (1989) Synthesis and biological properties of purine and pyrimidine 5′-deoxy-5′-dihydroxyphosphinyl-β-D-ribofuranosyl analogs of AMP, GMP, IMP, and CMP. J Med Chem 32: 1307–1313
Selgrade MK, Osborn JE (1973) Divergence of mouse brain interferon response following virulent or avirulent Newcastle disease virus inoculation. Proc Soc Exp Biol Med 143: 12–18
Selgrade MK, Nedrud JG, Collier AM, Gardner DE (1981) Effects of cell source, mouse strain, and immunosuppressive treatment on production of virulent and attenuated murine cytomegalovirus. Infect Immun 33: 840–847
Smee DF, Martin JC, Verheyden JPH, Matthews TR (1983) Anti-herpesvirus activity of the acyclic nucleoside 9-(1,3-dihydroxy-2-propoxymethyl)guanine. Antimicrob Agents Chemother 23: 676–682
Smee DF, McKernan PA, Alaghamandan HA, Frank KB, Ramasamy K, Revankar GR, Robins RK (1988) Antiviral activities of 2′-deoxyribofuranosyl and arabinofuranosyl analogs of sangivamycin against retro- and DNA viruses. Antiviral Res 10: 263–278
Smee DF, McKernan PA, Nord LD, Willis RC, Petrie CR, Riley TM, Revankar GR, Robins RK, Smith RA (1987) Novel pyrazolo[3,4-d]pyrimidine nucleoside analog with broad-spectrum antiviral activity. Antimicrob Agents Chemother 31: 1535–1541
Sommadossi J-P, Carlisle R (1987) Toxicity of 3′-azido-3′-deoxythymidine and 9-(1,3-dihydroxy-2-propoxymethyl)guanine for normal human hematopoietic progenitor cells in vitro. Antimicrob Agents Chemother 31: 452–454
Wingard JR, Stuart RK, Saral R, Burns WH (1981) Activity of trifluorothymidine against cytomegalovirus. Antimicrob Agents Chemother 20: 286–290
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Smee, D.F., Colletti, A., Alaghamandan, H.A. et al. Evaluation of continuous cell lines in antiviral studies with murine cytomegalovirus. Archives of Virology 107, 253–260 (1989). https://doi.org/10.1007/BF01317921
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01317921