Transformation of non-lymphoid cells by herpesviruses: a review.
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Biomedical subjects
Publications and source records attributed to F Rapp.
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We have made use of a novel filter hybridization approach in order to map the herpes simplex virus (HSV) DNA sequences which are present in three HSV thymidine kinase (TK)-transformed cell lines. The cell line 33A+ which was produced by infection of 3T3 TK- cells with UV-irradiated HSV-2 (333) was found to contain one contiguous stretch of viral DNA sequences which maps between 0.15 and 0.57 on the HSV-2 genome. The sequences mapping from 0.31 to 0.37 were present in 3--4-fold higher abundance than the rest of the viral DNA sequences in this cell line. Cell lines 5A and 8N were produced by transfection of mouse CL1D cells with sheared HSV-1 (1023) DNA. The cell line 5A was found to contain a contiguous set of viral DNA sequences mapping between 0.26 and 0.41 on the HSV-1 genome. The cell line 8N was found to contain three non-contiguous sets of viral DNA sequences, mapping between 0.09 and 0.41, 0.53 and 0.58, and 0.94 and 1.0 on the HSV-1 genome. These results seem to indicate that many different sets of viral DNA sequences can be incorporated into the cell during HSV-mediated biochemical transformation.
The DNA of varicella-zoster virus (VZV) has been characterized by sucrose gradient sedimentation, restriction enzyme cleavage with either EcoRI or HindIII site-specific endonucleases, and by isopycnic banding in caesium chloride. Comparisons of the DNAs from different clinical isolates have been made. DNAs from VZVs isolated from either varicella or herpes zoster are indistinguishable on the basis of size and restriction enzyme cleavage pattern. The buoyant density in caesium chloride of the DNA of VZV isolated from varicella was reproducibly slightly lower than that of the DNA of VZV isolated from herpes zoster.
Diverse alterations in biological properties of CMV-transformed cell lines were observed during prolonged in vitro cultivation. In the CMV-Mj-HEL-2 parent line there was a gradual decrease in the number of cells expressing CMV-related antigens; at the same time, an increase in oncogenicity was observed. One tumour line, designated CMV-Mj-HEL-2,T-1, however, retained the original ratio of cells expressing CMV-related antigens for over 100 in vitro passages. The cells lost their original moderate oncogenicity during this period. A later increase in the ratio of cells without CMV antigenic markers was accompanied by the return of moderate tumorigenicity and karyotypic changes. Both cell lines were studied to determine sensitivity to superinfection with herpesviruses, induction of immune response in nude mice, and release of infectious virus.
The thymidine analogue, 1-beta-arabinofuranosylthymine (ara-T), has previously been found to selectively inhibit herpes simplex virus (HSV) replication. At a relatively non-toxic concentration (50 microgram/ml), ara-T reduced HSV yields by a factor of 10,000-100,000. Ara-T was also effective in inhibiting the replication of variecella-zoster virus (VZV) in vitro in human embryo fibroblasts, completely preventing VZV-specific cytopathic effects (CPE). Ara-T reduced the cell-free virus and plaque-forming cell (PFC) yields of VZV as well as of the simian varicella-like virus, Delta herpesvirus. In contrast to HSV and VZV, cytomegalovirus (CMV) replication was relatively resistant to ara-T. Neither CPE nor the incorporation of 3H-thymidine into acid-insoluble material in CMV-infected cells was markedly affected. Interpretation of these results with regard to virus-induced deoxypyrimidine kinase is discussed.
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The etiology of cancer resembles that of many other diseases in that multiple factors may be required. Because of this, the role or viruses in the etiology of human cancers is especially difficult to assess. When animal tumor systems were used as models, the roles of various predisposing characteristics in virus oncogenesis were elucidated. Extrapolation of these findings to the human diseases suggests the importance of genetics, age, hormones, immune competence, and stress in determining susceptibility to tumor development in individuals infected with an oncogenic virus. The importance of cofactors in induction of those human tumors most strongly associated with virus infection, including Burkitt's lymphoma, nasopharyngeal carcinoma, cerviccal carcinoma, acute myelogenous leukemia, and breast cancer, is reviewed. Understanding of the role of these cofactors in virus carcinogenesis may lead to disease prevention through elimination of one or more of the cofactors.
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A procedure is described which yields a significant percentage of long-term mixed cell cultures of human prostatic tissue. Attempts were made to suppress the proliferation of stromal fibroblasts and to characterize the cultured cells as those of prostatic origin. The problems associated with establishing epithelial cell lines are discussed.
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A human genital isolate of cytomegalovirus is shown to have transformed human embryonic lung cells in vitro. These cells produce tumors when injected into athymic nude mice. Two cell lines derived from tissue from human prostatic carcinoma have survived more than 20 passages in vitro and demonstrate cytomegalovirus-specific membrane antigen. Significant humoral antibody titers against cytomegalovirus have been demonstrated. Cell-mediated lymphocytotoxicity against these transformed cells has been demonstrated in patients with urinary tract tumors. This evidence indicates that an association between cytomegalovirus and human prostatic cancer may be more than coincidental.
Lymphocytes from patients with urologic cancer were tested in microcytotoxicity assays against human cells transformed by cytomegalovirus. Human lymphocytes were significantly cytotoxic against the transformed cell line when compared to a normal human control cell line. Patients with prostatic carcinoma demonstrated greater target cell reduction than those with benign prostatic hyperplasia.
Ten polypeptides were detected on the surface of the virion of herpes simplex virus type 1. Of these ten polypeptides, three were detected in hamster cells transformed by herpes simplex type 1.