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Cellular mutations and drug resistance probed by herpes simplex virus
Author(s) -
Campisi Judith,
Pardee Arthur B.
Publication year - 1981
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041090313
Subject(s) - herpes simplex virus , thymidine kinase , biology , cell culture , virology , virus , fibroblast , virus quantification , microbiology and biotechnology , thymidine , viral replication , immunofluorescence , in vitro , immunology , biochemistry , antibody , genetics
The growth of herpes simplex virus type 1 (HSV‐1), monitored by plaque assay, was inhibited by the cellular antimetabolites thioguanine (TG), cytosine arabinoside (AraC), methotrexate (MTX), and 5‐fluorodeoxyuridine (5‐FUdR). These results suggested a rapid means for assaying cellular drug sensitivity, based on the ability of infected cells to support viral replication. We have explored the feasibility of a virus‐mediated assay for cellular metabolic function in two model systems. Using an immunofluorescence assay to assess viral growth, we found that all of the antimetabolites tested were effective in diminishing HSV‐1 specific fluorescence in human fibroblasts. However, a DNA‐damaging agent, bleomycin, was lethal to cells but was completely ineffective in reducing viral fluorescence. HSV‐1 growth was markedly decreased by TG in a normal human fibroblast strain, FS‐2. In contrast, a Lesch‐Nyhan strain (LNF), resistant to TG owing to its genetic defect, showed no suppression of viral growth in the presence of TG. The drug's effect on viral fluorescence closely paralleled its effect on cellular colony forming ability and rate of cellular DNA synthesis. Thymidine kinase‐deficient HSV‐1 (TK − HSV‐1) did not grow in a normal mouse fibroblast line (A31) in the presence of 5‐FUdR. However, a TK − derivative of the A31 line allowed full production of the TK − HSV‐1 antigens at low to moderate doses of 5‐FUdR. Two potential applications for this assay are the prenatal diagnosis of some genetic disorders and the rapid detection of drug resistant populations in tumor specimens. Toward these ends, we demonstrated that human fibroblasts from patients with the hereditary disorder Xeroderma pigmentosum (group A) were easily distinguished from normal human fibroblasts by their inability to support the growth of UV‐irradiated HSV‐1. We also investigated the effects of TG upon HSV‐1 fluorescence in two human tumor cell lines isolated from head and neck squamous cell carcinomas (SCC‐15) and (SCC‐25). Whereas TG was effective in reducing viral fluorescence in SCC‐15 cells, it was only marginally so in SCC‐25 cells. These latter cells showed the greater resistance to TG by growth and isotope incorporation experiments.