[Herpesvirus and human cancer].
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Biomedical subjects
Publications and source records attributed to F Rapp.
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A new human endometrial cell line, SCRC-1, has been established from a uterine adenocarcinoma. During a period of 57 weeks in culture, the SCRC-1 cells have been passaged over 100 times. Cultures have an epitheloid morphology, high mitotic activity, and a tendency to form multiple cell layers. The cells are polygonal and are arranged in pavement-like fashion. Most have multiple nucleoli, some are multinucleated, and all have heavily vacuolated cytoplasm. Pretreatment with iododeoxyuridine did not reveal viral markers by cocultivation or immunofluorescence techniques. Karyologic studies indicate that the SCRC11 cell line is essentially diploid with a small subpopulation of tetraploid cells. Over a period of 20 weeks, 1 of 14 athymic nude mice inoculated with SCRC-1 cells developed a tumor which histologically closely resembled the original human tumor. Cells were subsequently propagated from this tumor and the resulting epitheloid-like cell line was designated SCRC-1, Tu 1.
The incidence of trigeminal ganglion infection after corneal inoculation of guinea pigs with thymidine kinase-negative mutants of herpes simplex virus was markedly reduced compared to infection after inoculation of thymidine kinase-positive virus. Thymidine kinase-negative herpes simplex virus replicated well in ocular tissues in which dividing or potentially dividing cells were present, but not in trigeminal ganglion infection of nondividing neurons. Thymidine kinase-positive virus, however, replicated well in ocular tissues as well as in trigeminal ganglion. These results suggest that thymidine kinase expression of herpes simplex virus may be important in infections of sensory ganglia.
Enhancement of host cell reactivation (HCR) of ultraviolet (UV)-irradiated herpes simplex virus (HSV) was demonstrated in cell cultures pretreated with caffeine, hydroxyurea, or 5-bromodeoxyuridine (BrdUrd). The effect of caffeine on HCR was shown to depend on the time of drug treatment with respect to infection. In cultures treated with caffeine during the course of virus replication, the infectivity of irradiated HSV was reduced about nine-fold, while cultures pretreated with the drug before infection showed an increase in infectivity. The extent of HCR enhancement depended on the time interval between treatment with caffeine and infection, drug concentration, and the UV irradiation dose to which HSV was exposed. Magnitude of enhancement of HCR by caffeine differed in various cell species. The results suggest that enhanced HCR of UV-irradiated HSV by DNA antimetabolities is associated with DNA repair activated in consequence of cell DNA damage.
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Biochemical and genetic methods have been used to investigate a defective variant of measles virus previously isolated from a patient with subacute sclerosing panencephalitis (SSPE). Since its isolation, this syncytiogenic strain (SSPE-BIKEN) has remained cell-associated; infected cells do not hemadsorb and do not release infectious virus. Immune precipitation and polyacrylamide gel electrophoresis were used to study the synthesis of measles virion proteins in SSPE-BIKEN-infected cells. All of the virion proteins were detected in immune precipitated whole cell extracts. However, the hemagglutinin (HA) protein was not detected on the cell surface by lactoperoxidase iodination. These results suggest that the failure of the HA protein to insert into the cell membrane accounts for the block in the release of infections virus. Radioactively labeled, noninfectious, virus-like particles have been purified from the media of SSPE-BIKEN-infected cells. These particles contain virus nucleocapsid, nucleocapsid-associated, and membrane proteins, but very little HA and hemolysin proteins. Genetic complementation between SSPE-BIKEN and a temperature-sensitive mutant of measles virus was observed and suggests that the SSPE isolate defect is due to a mutation. Additional evidence of a mutation is provided by the detection of low frequency revertant progeny in SSPE-BIKEN stocks. Our results support the hypothesis that genetic variants of measles virus are involved in the etiology of SSPE.
The ability of human cytomegalovirus to stimulate replication of herpes simplex virus type 2 (HSV-2) was examined. The system used involved HSV-2-infected human embryonic lung cells under conditions (39.5-40 degrees C) in which HSV-2 remains undetectable. Reactivation of HSV-2 was maximal and persisted for the longest duration when cultures were superinfected with 0.02 plaque-forming unit of human cytomegalovirus per cell. Infectious HSV-2 appeared 2 days after superinfection with human cytomegalovirus and ranged from 10(2) to 10(6) plaque-forming units per culture. Virus reactivated from these cultures was neutralized by rabbit immune serum produced against HSV-2. The specificity of this interaction was demonstrated by various criteria: production of HSV-2 was not observed in cultures treated with mock infecting fluid, and inactivation of human cytomegalovirus by heat, ultraviolet irradiation, or immune serum prior to superinfection eliminated its ability to induce HSV-2 replication. These results sugges that interaction between these two human herpesviruses may be of importance in herpesvirus latency in vivo.
The guanine derivative 9-(2-hydroxyethoxymethyl) guanine (acycloguanosine), a potent inhibitor of herpes simplex virus (HSV) multiplication, was found to have marked anticellular activity against HSV-transformed thymidine kinase-positive cells. HSV type I-transformed cells were more sensitive to the drug than HSV type 2-transformed cells.
The antiviral effect of the nucleoside analog 5-iodo-5'-amino-2',5'-dideoxyuridine (AIU) was tested with three isolates of varicella-zoster virus (VZV). AIU concentrations of 10 to 800 muM (3.5 to 288 mug/ml) reduced the number of plaques produced by VZV-infected cells and cell-free VZV from approximately 30 to 95%. Smaller plaque size was also observed in the presence of AIU. AIU was less effective than arabinofuranosylthymine in reducing VZV-induced plaques since as little as 5 mug of arabinofuranosylthymine per ml completely blocked plaque formation by cell-free VZV. Toxicity assays with human diploid embryo fibroblast cells were also carried out. Drug concentrations as high as 800 muM were not toxic to human diploid embryo fibroblast cells as determined by radiolabeling of cell deoxyribonucleic acid, ribonucleic acid, and protein.
Hamster embryo fibroblasts persistently infected with a derivative of the Schwarz vaccine strain of measles virus spontaneously released virus particles with an average buoyant density considerably lower than that of the parental virus. The released virus contained all of the measles virus structural proteins and interfered with replication of standard virus. All of the virus structural proteins were associated with a membrane-free cytoplasmic extract from the persistently infected cells. Membrane-free cytoplasmic extracts prepared from Vero cells lytically infected with Schwarz strain measles contained little or no virus envelope structural protein. Maintenance of persistent infection may involve both the presence of virus variants and a defect in the ability of the infected cell to replicate the virus efficiently.
Plasminogen activator was produced by both human embryo fibroblasts (a permissive system) and hamster embryo fibroblasts (a nonpermissive system) after exposure to human cytomegalovirus. The level of this activator was measured by using plates coated with [125I]fibrin. The production of plasminogen activator was enhanced when the human cells were exposed to human cytomegalovirus previously irradiated with UV light (5,520 to 55,200 ergs/mm2).
About 1% of Raji cells showed sensitivity to herpes simplex virus type 2 (HSV-2) infection when tested by infectious center assays or immunofluorescence tests, and the percentage did not change during cell passage. The addition of hydrocortisone to Raji cells persistently infected with HSV-2 caused a marked increase in virus production and in the number of HSV-producing cells. In the case of HSV-1, it was shown that the addition of hydrocortisone was required to maintain persistent infection. These observations suggest that control of replication of HSV-1 and HSV-2 in these cells is regulated by different mechanisms.
The 32P-labeled DNAs from seven different clinical isolates of human varicella-zoster virus (VZV) were independently digested with five site-specific restriction endonucleases, EcoRI, HindIII, SmaI, BamHI, and AvaI. The digestion products were analyzed by electrophoresis on 0.5% agarose gels followed by autoradiography of the dried gels. Evaluation of the restriction enzyme cleavage patterns revealed small variations among the VZV DNAs. The VZV DNAs were also compared based on their buoyant densities in CsCl. No significant buoyant density differences were detected among the VZV DNAs.
Human embryo fibroblasts treated with diethylstilbestrol (DES) gave rise to slightly increased plaque production by cytomegalovirus (CMV): plaques were also slightly larger in treated cells when compared to those in untreated cultures. There was no significant enhancement of plaque production by herpes simplex virus (HSV) types 1 and 2 in the human cells or in primary rabbit kidney cells treated with DES. Growth analyses of HSV and CMV in DES-treated cells failed to reveal enhancement of virus production when compared to untreated cells. The frequency of biochemical transformation of mouse cells deficient in thymidine kinase (TK) to the TK+ phenotype by HSV was markedly enhanced when cells were pretreated with DES. TK+-transformed colonies arising from DES-treated cells were also larger than those derived from untreated cultures. These reveal that DES increases susceptibility of human cell lines to CMV infection and enhances efficiency of biochemical transformation of mouse cells by HSV.
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A quantitative assay for the biochemical transformation of thymidine kinaseless mouse cells (N cIA cI10 cells) to an enzyme-positive phenotype by herpes simplex virus has been used to examine the effect of the carcinogen nitrosomethylurea on cell transformation. Exposure of cells to the chemical carcinogen, followed by virus infection, resulted in enhancement of transformation when compared to that seen with virus or chemical alone. Enhancement occurred after doses of nitrosomethylurea that were nontoxic to treated cells. A strong time dependence after treatment was demonstrated for enhancement which was correlated with the presence and excision-repair of alkylated DNA in treated cells.