Bovine herpesvirus-4 (BHV-4), a gammaherpesvirus lacking a clear disease association, productively infects multiple cell lines of various species and causes cell death. A human rhabdomyosarcoma cell line, RD-4, infected with BHV-4 produced low levels of early and late viral RNAs and infectious virus, but exhibited no cytopathic effect. Using a recombinant BHV-4 containing a neomycin-resistance gene, we established RD-4-derived cell lines persistently infected with BHV-4. The viral genome in these cells was predominantly circular. Because of drug selection, every cell contained a viral genome. In addition, all cells stained with a BHV-4-specific antiserum. Therefore, these cell lines are not carrier cultures. These cells produced infectious virus at all passages tested. Even though cells were selected and maintained at a concentration of geneticin at least 2·5 times that necessary to kill uninfected RD-4 cells, selected cells contained only approximately one viral genome per diploid host cell genome. Persistently infected cells grew more slowly than uninfected cells, even in the absence of drug. The slower growth of these cells suggests that any growth advantage conferred by multiple copies of the neomycin-gene-carrying viral genome might be offset by the detrimental effects of viral gene expression. This situation contrasts with other gammaherpesviruses, which are able to growth-transform cells.
Ballestas, M. E., Chatis, P. A. & Kaye, K. M. (1999). Efficient persistence of extrachromosomal KSHV DNA mediated by latency-associated nuclear antigen.Science284, 641-644.[CrossRef][Google Scholar]
Bartha, A., Fadol, A. M., Liebermann, H., Ludwig, H., Mohanty, S. B., Osorio, F. A., Reed, D. E., Storz, J., Straub, O. C., Van der Maaten, M. J. & Wellermans, G. (1987). Problems concerning the taxonomy of the ‘Movar-Type’ bovine herpesvirus.Intervirology28, 1-7.[CrossRef][Google Scholar]
Bataille, D. & Epstein, A. (1994). Herpes simplex virus replicative concatemers contain L components in inverted orientation.Virology203, 384-388.[CrossRef][Google Scholar]
Bermudez-Cruz, R., Zhang, L. & van Santen, V. L. (1997). Characterization of an abundant, unique 1·7-kilobase bovine herpesvirus 4 (BHV-4) late RNA and mapping of a BHV-4 IE2 transactivator-binding site in its promoter-regulatory region.Journal of Virology71, 527-538.
[Google Scholar]
Broll, H., Buhk, H.-J., Zimmermann, W. & Goltz, M. (1999). Structure and function of the prDNA and the genomic termini of the γ2-herpesvirus bovine herpesvirus type 4.Journal of General Virology80, 979-986.
[Google Scholar]
Cesarman, E., Moore, P. S., Rao, P. H., Inghirami, G., Knowles, D. M. & Chang, Y. (1995). In vitro establishment and characterization of two acquired immunodeficiency syndrome-related lymphoma cell lines (BC-1 and BC-2) containing Kaposi’s sarcoma-associated herpesvirus-like (KSHV) DNA sequences.Blood86, 2708-2714.
[Google Scholar]
Chang, L.-Y. & van Santen, V. L. (1992). Immediate-early, early, and late RNAs in bovine herpesvirus-4-infected cells.Virology191, 909-920.[CrossRef][Google Scholar]
Decker, L. L., Shankar, P., Khan, G., Freeman, R. B., Dezube, B. J., Lieberman, J. & Thorley-Lawson, D. A. (1996). The Kaposi sarcoma-associated herpesvirus (KSHV) is present as an intact latent genome in KS tissue but replicates in the peripheral blood mononuclear cells of KS patients.Journal of Experimental Medicine184, 283-288.[CrossRef][Google Scholar]
Desrosiers, R. C., Kamine, J., Bakker, A., Silva, D., Woychik, R. P., Sakai, D. D. & Rottman, F. M. (1985). Synthesis of bovine growth hormone in primates by using a herpesvirus vector.Molecular and Cellular Biology5, 2796-2803.
[Google Scholar]
Drexler, H. G., Uphoff, C. C., Gaidano, G. & Carbone, A. (1998). Lymphoma cell lines: in vitro models for the study of HHV-8+ primary effusion lymphomas (body cavity-based lymphomas).Leukemia12, 1507-1517.[CrossRef][Google Scholar]
Dubuisson, J., Boulanger, D., Bublot, M., Thiry, E. & Pastoret, P.-P. (1989a). Proteins specified by bovine herpesvirus type 4: structural proteins of the virion and identification of two major glycoproteins by using monoclonal antibodies.Journal of General Virology70, 1743-1753.[CrossRef][Google Scholar]
Dubuisson, J., Thiry, E., Bublot, M., Thomas, I., van Bressem, M. F., Coignoul, F. & Pastoret, P.-P. (1989b). Experimental infection of bulls with a genital isolate of bovine herpesvirus-4 and reactivation of latent virus with dexamethasone.Veterinary Microbiology21, 97-114.[CrossRef][Google Scholar]
Dubuisson, J., Pastoret, P.-P. & Thiry, E. (1991). Temporal control of bovine herpesvirus 4 glycoprotein synthesis.Journal of General Virology72, 1429-1434.[CrossRef][Google Scholar]
Egyed, L. (1998). Replication of bovine herpesvirus type 4 in human cells in vitro.Journal of Clinical Microbiology36, 2109-2111.
[Google Scholar]
Egyed, L. & Bartha, A. (1998). PCR studies on the potential sites for latency of BHV-4 in calves.Veterinary Research Communications22, 209-216.[CrossRef][Google Scholar]
Ehlers, B., Buhk, H.-J. & Ludwig, H. (1985). Analysis of bovine cytomegalovirus genome structure: cloning and mapping of the monomeric polyrepetitive DNA unit, and comparison of European and American strains.Journal of General Virology66, 55-68.[CrossRef][Google Scholar]
Flore, O., Rafii, S., Ely, S., O’Leary, J. J., Hyjek, E. M. & Cesarman, E. (1998). Transformation of primary human endothelial cells by Kaposi’s sarcoma-associated herpesvirus.Nature394, 588-592.[CrossRef][Google Scholar]
Galik, P. K., van Santen, V. L., Stringfellow, D. A., Bird, R. C., Wright, J. C. & Smith, P. C. (1993). Development of a DNA probe for identification of bovine herpesvirus 4.American Journal of Veterinary Research54, 653-659.
[Google Scholar]
Gardella, T., Medveczky, P., Sairenji, T. & Mulder, C. (1984). Detection of circular and linear herpesvirus DNA molecules in mammalian cells by gel electrophoresis.Journal of Virology50, 248-254.
[Google Scholar]
Grassmann, R. & Fleckenstein, B. (1989). Selectable recombinant herpesvirus saimiri is capable of persisting in a human T-cell line.Journal of Virology63, 1818-1821.
[Google Scholar]
Huang, E.-S. (1975). Human cytomegalovirus. IV. Specific inhibition of virus-induced DNA polymerase activity and viral DNA replication by phosphonoacetic acid.Journal of Virology16, 1560-1565.
[Google Scholar]
Jung, J. U., Choi, J. K., Ensser, A. & Biesinger, B. (1999). Herpesvirus saimiri as a model for gammaherpesvirus oncogenesis.Seminars in Cancer Biology9, 231-239.[CrossRef][Google Scholar]
Keil, G. M., Heinze, A. & Mächtig, B. (1990). Bovine herpesvirus 4 vector for life-virus vaccines?Zentralblatt für Bakteriologie272, 375.
[Google Scholar]
Kieff, E. (1996). Epstein–Barr virus and its replication. In Fields Virology, pp. 1109-1162. Edited by B. N. Fields, D. M. Knipe & P. M. Howley. Philadelphia: Lippincott–Raven Publishers.
Krogman, L. A. & McAdaragh, J. P. (1982). Recrudescence of bovine herpesvirus-5 in experimentally infected calves.American Journal of Veterinary Research43, 336-338.
[Google Scholar]
Kung, S. H. & Medveczky, P. G. (1996). Identification of a herpesvirus saimiri cis-acting DNA fragment that permits stable replication of episomes in transformed T cells.Journal of Virology70, 1738-1744.
[Google Scholar]
Lomonte, P., Bublot, M., van Santen, V., Keil, G., Pastoret, P. P. & Thiry, E. (1996). Bovine herpesvirus 4: genomic organization and relationship with two other gammaherpesviruses, Epstein–Barr virus and herpesvirus saimiri.Veterinary Microbiology53, 79-89.[CrossRef][Google Scholar]
Lupton, S. & Levine, A. J. (1985). Mapping genetic elements of Epstein–Barr virus that facilitate extrachromosomal persistence of Epstein–Barr virus-derived plasmids in human cells.Molecular and Cellular Biology5, 2533-2542.
[Google Scholar]
Miller, G. (1990). Epstein–Barr virus. In Virology, pp. 1921-1958. Edited by B. N. Fields, D. M. Knipe, R. M. Chanock, M. S. Hirsch, J. L. Melnick, T. P. Monath & B. Roizman. New York: Raven Press.
Moore, P. S. & Chang, Y. (1998). Kaposi’s sarcoma-associated herpesvirus-encoded oncogenes and oncogenesis.Journal of the National Cancer Institute Monograph Series23, 65-71.
[Google Scholar]
Moses, A. V., Fish, K. N., Ruhl, R., Smith, P. P., Strussenberg, J. G., Zhu, L., Chandran, B. & Nelson, J. A. (1999). Long-term infection and transformation of dermal microvascular endothelial cells by human herpesvirus 8.Journal of Virology73, 6892-6902.
[Google Scholar]
Naeem, K., Caywood, D. D., Goyal, S. M., Werdin, R. E. & Murtaugh, M. P. (1993). Persistence of bovid herpesvirus-4 in experimentally inoculated pregnant rabbits.Microbiologica16, 87-94.
[Google Scholar]
Nilsson, K. (1979). The nature of lymphoid cell lines and their relationship to the virus. In The Epstein–Barr Virus, pp. 225-281. Edited by M. A. Epstein & B. G. Achong. Berlin: Springer-Verlag.
Osorio, F. A. & Reed, D. E. (1983). Experimental inoculation of cattle with bovine herpesvirus-4: evidence for a lymphoid-associated persistent infection.American Journal of Veterinary Research44, 975-980.
[Google Scholar]
Osorio, F. A., Reed, D. E. & Rock, D. L. (1982). Experimental infection of rabbits with bovine herpesvirus-4: acute and persistent infection.Veterinary Microbiology7, 503-513.[CrossRef][Google Scholar]
Osorio, F. A., Rock, D. L. & Reed, D. E. (1985). Studies on the pathogenesis of a bovine cytomegalo-like virus in an experimental host.Journal of General Virology66, 1941-1951.[CrossRef][Google Scholar]
Peterson, R. B. & Goyal, S. M. (1988). Propagation and quantitation of animal herpesviruses in eight cell culture systems.Comparative Immunology, Microbiology and Infectious Diseases11, 93-98.[CrossRef][Google Scholar]
Poffenberger, K. M. & Roizman, B. (1985). A noninverting genome of a viable herpes simplex virus 1: presence of head-to-tail linkages in packaged genomes and requirements for circularization after infection.Journal of Virology53, 587-595.
[Google Scholar]
Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989).Molecular Cloning: A Laboratory Manual., 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
Severini, A., Morgan, A. R., Tovell, D. R. & Tyrrell, D. L. (1994). Study of the structure of replicative intermediates of HSV-1 DNA by pulsed-field gel electrophoresis.Virology200, 428-435.[CrossRef][Google Scholar]
Simmer, B., Alt, M., Buckreus, I., Berthold, S., Fleckenstein, B., Platzer, E. & Grassmann, R. (1991). Persistence of selectable herpesvirus saimiri in various human haematopoietic and epithelial cell lines.Journal of General Virology72, 1953-1958.[CrossRef][Google Scholar]
Sugden, B., Phelps, M. & Domoradzki, J. (1979). Epstein–Barr virus DNA is amplified in transformed lymphocytes.Journal of Virology31, 590-595.
[Google Scholar]
Thiry, E., Bublot, M., Dubuisson, J. & Pastoret, P.-P. (1989). Bovine herpesvirus-4 (BHV-4) infections of cattle. In Herpesvirus Diseases of Cattle, Horses, and Pigs, pp. 96-115. Edited by G. Wittmann. Boston: Kluwer Academic Publishers.
Truman, D., Ludwig, H. & Storz, J. (1986). Bovine herpesvirus type 4: studies on the biology and spread in cattle herds and in insemination bulls.Journal of Veterinary Medicine, Series B33, 485-501.[CrossRef][Google Scholar]
Tsygankov, A. Y. & Romano, G. (1999). Mechanisms of cell transformation by Herpesvirus saimiri.Anticancer Research19, 973-983.
[Google Scholar]
Usherwood, E. J., Stewart, J. P. & Nash, A. A. (1996). Characterization of tumor cell lines derived from murine gammaherpesvirus-68-infected mice.Journal of Virology70, 6516-6518.
[Google Scholar]
van Santen, V. L. (1991). Characterization of the bovine herpesvirus 4 major immediate-early transcript.Journal of Virology65, 5211-5224.
[Google Scholar]
van Santen, V. L. & Chang, L.-Y. (1992). Cloning and mapping of EcoRI, HindIII, and PstI fragments of bovine herpesvirus 4 (DN-599).Intervirology34, 44-52.
[Google Scholar]
Vanderplasschen, A., Goltz, M., Lyaku, J., Benarafa, C., Buhk, H. J., Thiry, E. & Pastoret, P. P. (1995). The replication in vitro of the gammaherpesvirus bovine herpesvirus 4 is restricted by its DNA synthesis dependence on the S phase of the cell cycle.Virology213, 328-340.[CrossRef][Google Scholar]
Yates, J., Warren, N., Reisman, D. & Sugden, B. (1984). A cis-acting element from the Epstein–Barr viral genome that permits stable replication of recombinant plasmids in latently infected cells.Proceedings of the National Academy of Sciences, USA81, 3806-3810.[CrossRef][Google Scholar]
Yates, J. L., Warren, N. & Sugden, B. (1985). Stable replication of plasmids derived from Epstein–Barr virus in various mammalian cells.Nature313, 812-815.[CrossRef][Google Scholar]
Zhang, X., Efstathiou, S. & Simmons, A. (1994). Identification of novel herpes simplex virus replicative intermediates by field inversion gel electrophoresis: implications for viral DNA amplification strategies.Virology202, 530-539.[CrossRef][Google Scholar]