Viral Encephalomyelitis of Pigeons. IV. Growth of the Virus in Tissue Cultures
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Growth and cytopathogenicity of pigeon herpes encephalomyelitis virus (PHEV) in avian and mammalian cell cultures were investigated. The virus was cytopathogenic to all avian primary cell cultures tested and produced large syncytia with intranuclear inclusions. Viral antigen was detected in the nuclei of infected cells 6 hr postinoculation. Infective virus, however, was obtained 8 hr post-inoculation. Maximum virus yields in avian cell cultures were reached 72 hr postinoculation. In mammalian cell lines tested, the virus proved to be cytopathogenic except in swine embryo kidney cell lines. The cytopathic effect in mammalian cell lines was characterized by the rounding and clumping of cells., Moderate virus yields were obtained with lamb kidney and bovine embryo thymus cell lines, but not with other cell lines tested. Growth behavior of the virus in cell cultures in comparison with other human and avian herpesviruses is discussed.Keywords:
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SummaryVaricella-zoster virus was shown to replicate and to cause a cytopathic effect in guinea pig embryo cell culture, characterized by syncytia and intranuclear inclusions. Small amounts of cell-free virus were released from the cells, which also produced CF antigen. Electron microscopy showed herpes-type virus particles in the infected cells.
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Rinderpest virus
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The outbreak of sialoadenitis occurred in a laboratory rat colony and the causative agent was isolated from the affected salivary glands of diseased rats using the established cell line LBC. The isolate readily multiplied, producing clear cytopathic effects with syncytium formation, and it was identified virologically and serologically as rat sialodacryoadenitis virus. In attempts to isolate the virus by primary rat kidney (PRK) cells and suckling mice as well as LBC cells, the LBC cells showed higher susceptibility for the virus growth as compared with PRK cells or the brain of suckling mice. The isolation rate of virus was 100% (5/5) in LBC, 40% (2/5) in PRK cells and 60% (3/5) in suckling mice. After four passages in the LBC cells, the virus did not produce disease in adult rats, while the mouse brain-passaged virus did.
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Hinze, Harry C. (University of Wisconsin, Madison), and Duard L. Walker . Variation of herpes simplex virus in persistently infected tissue cultures. J. Bacteriol. 82: 498–504. 1961.—Cultures of human conjunctiva, HeLa, and KB cells infected with herpes simplex virus (HF strain) were cultured for prolonged periods in medium containing low levels of antibody. After continuous culture of the infected cell lines for 6 months, two major changes were noted in the character of the virus present in the cultures. These changes consisted of an alteration in the type of cytopathic effect produced by the virus and marked loss of virulence for mice. Further study of the virus from the cultures revealed that it was still antigenically similar to the original strain of herpes virus. It was found that the variant produced an altered, proliferative type of cytopathic effect only in the presence of herpes antibody. The variant multiplied more slowly in human conjunctiva cells in culture than did the parent virus, and the variant had lost practically all capacity for multiplication in the brain of mice.
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Growth and cytopathogenicity of pigeon herpes encephalomyelitis virus (PHEV) in avian and mammalian cell cultures were investigated. The virus was cytopathogenic to all avian primary cell cultures tested and produced large syncytia with intranuclear inclusions. Viral antigen was detected in the nuclei of infected cells 6 hr postinoculation. Infective virus, however, was obtained 8 hr post-inoculation. Maximum virus yields in avian cell cultures were reached 72 hr postinoculation. In mammalian cell lines tested, the virus proved to be cytopathogenic except in swine embryo kidney cell lines. The cytopathic effect in mammalian cell lines was characterized by the rounding and clumping of cells., Moderate virus yields were obtained with lamb kidney and bovine embryo thymus cell lines, but not with other cell lines tested. Growth behavior of the virus in cell cultures in comparison with other human and avian herpesviruses is discussed.
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The LFBK-α v β 6 cell line is highly sensitive for the isolation of foot-and-mouth disease virus (FMDV) and porcinophilic vesicular viruses. However, LFBK-α v β 6 cells are contaminated with a non-cytopathic bovine viral diarrhea virus (BVDV), which complicates handling procedures in areas where other cell lines are maintained, as well downstream use of viral isolates. In this study, we used an aromatic cationic compound (DB772) to treat LFBK-α v β 6 cells using an approach that has been previously used to eliminate persistent BVDV from fetal fibroblast cell lines. After three cell passages with 4 μM DB772, BVDV could no longer be detected in unclarified cell suspensions using a pan-pestivirus real-time RT-PCR assay, and remained undetectable after treatment was stopped (nine passages) for an additional 28 passages. The analytical sensitivity of the DB772-treated LFBK-α v β 6 cultures (renamed WRL-LFBK-α v β 6 ) to titrations of FMDV and other vesicular virus isolates was comparable to untreated LFBK-α v β 6 cells. These new BVDV-free cells can be handled without the risk of cross-contaminating other cells lines or reagents, and used for routine diagnostics, in vivo studies and/or preparation of new vaccine strains.
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The interaction between 2 strains of group A streptococci in L-forms and the cells of the continuous cell lines HEp, HeLa, L, GPK and PEK, as well as the cells of primary human and chick embryo cell cultures was studied under conditions of infection with different doses. In most of the cell cultures used in this study L-form of streptococci showed no pronounced cytopathic effect. They could be isolated, when using cell cultures as inoculum, from the cultivated cells of continuous cell lines during the whole period of the cultivation of the infected monolayer (6--7 days), primary human embryo cell culture up to days 8--11 and from chick embryo cell culture up to days 1--3. In the cells of the continuous cell lines the maximum amount of L-forms was revealed on days 2--3 by the immunofluorescent technique. The ultrathin sections of HEp and HeLa cells infected with L-forms of streptococci were found to contain small elements similar to L-forms inside the cells and on their surface, which was not detected in the infected primary cell cultures.
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XC cells, derived from a Rous sarcoma virus-induced Wistar strain rat tumor, form syncytia when cultured in the presence of murine leukemia virus-producing mouse cells. However, one XC cell culture (designated as XC-v cells), found to produce type C virus particles, fails to form syncytia in the presence of murine leukemia virus-producing mouse cells. Coculture of XC-v cells and XC cells negative for type C virus particles leads to a moderate degree of syncytium formation. Infection of XC cells with either the Moloney (M) strain of mouse leukemia virus or type C virus particles released by XC-v cells results in the loss of ability of XC cells to form syncytia in the mixed culture cytopathogenicity test. The syncytium-forming ability of XC cells, therefore, is altered by the presence of a type C virus in these cells.
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Aedes albopictus
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