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>Hereditary orotic aciduria, Lesch‐Nyhan syndrome, and xeroderma pigmentosum probed by herpes simplex virus:125I‐Iododeoxycytidine incorporation as an assay for viral growth
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Hereditary orotic aciduria, Lesch‐Nyhan syndrome, and xeroderma pigmentosum probed by herpes simplex virus:125I‐Iododeoxycytidine incorporation as an assay for viral growth
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机译：Hereditary orotic aciduria, Lesch‐Nyhan syndrome, and xeroderma pigmentosum probed by herpes simplex virus:125I‐Iododeoxycytidine incorporation as an assay for viral growth
Abstract125I‐Iododeoxycytidine (125IdC) incorporation into acid‐insoluble material was a sensitive, rapid, and quantitative assay for the growth of herpes simplex virus type 1 (HSV‐1) in human fibroblasts. Cellular utilization of the isotope was 10 to 25 of the incorporation by infected cells and could be 80 inhibited by tetrahydrouridine (THU). Viral utilization was inhibited by acycloguanosine, thioguanine (TG), and cytosine arabinoside. Isotope was incorporated equally well by growing or quiescent infected cells. HSV‐1 was used to probe the metabolic capabilities of three mutant human fibroblast strains.125IdC incorporation quantitatively measured the ability of the virus to grow in these cells. Viral125IdC incorporation was sensitive to TG in normal fibroblasts but showed a 8‐ to 10‐fold greater resistance to TG in fibroblasts derived from patients with Lesch‐Nyhan syndrome (LN). Similarly, the growth of ultraviolet irradiated HSV‐1 in normal fibroblasts, as judged by125IdC incorporation, was 5‐fold greater than in fibroblasts derived from patients with xeroderma pigmentosum. In fibroblasts derived from patients with hereditary orotic aciduria, viral125IdC incorporation was sensitive to adenosine (AD) at concentrations which were slightly stimulatory in normal fibroblasts. This was a 2‐fold difference in AD sensitivity, which the radioassay reliably and quantitatively documented. Previous attempts to quantify and characterize this difference by immunofluorescence were unsuccessful. HSV‐1 infected cells could be individually identified by their incorporated125IdC; such cells had blackened nuclei in autoradiograms prepared 12 hr after infection. Normal cells infected in the presence of TG had many fewer labeled nuclei than LN cells similarly infected in the presence of the drug. Thus, this viral assay can be a rapid and accurate probe of cellular function and has potential for the identification of mutant cells in amniocentesis samples or tumor speciments. Towards this end, we determined that the AD and TC dose responses of infected amniotic fluid cells closely paralleled that of the nor
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