Isolation and Characterization of Acyclovir-resistant Mutants of Herpes Simplex Virus

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Abstract
Rat embryo fibroblasts (REF) were transformed in vitro with photoinactivated herpes simplex virus. Low passage (7 to 10) HSV-transformed rat cells (t-REF-line G) produced multiple tumours in 49% of newborn rats with a latent period of 20 to 24 weeks. An in vitro cloning procedure for transformants in the uncloned t-REF-line G cells produced clonal lines which varied from non-oncogenic to clonal lines producing tumours with shorter latent periods (10 to 14 weeks) compared to uncloned cells. At passage 30, t-REF-line G-clone 1 cells produced rapidly growing tumours in 100% of the newborn rats with a latent period of only 2 to 3 weeks. Tumour cells (RFS 12-22-75) established in culture produced tumours within 2 weeks after subcutaneous (s.c.) inoculation of weanling rats (100% with tumours) and they were transplantable to 100% of inoculated adult rats. Histopathological examination of all tumours produced in newborn, weanling or adult rats revealed large, poorly differentiated malignant fibrosarcomas; metastatic tumours were observed in the lungs of 10 to 20% of newborn rats inoculated s.c. with RSF cells. Approx. 25 to 50% of the clonal transformed or tumour cells synthesized HSV-specific-antigens detected by immunofluorescence. HSV-transformed and tumour cells are resistent to superinfection by the homologous transforming virus. Since the in vitro cloning procedure for transformant cells can readily segregate cells producing clonal lines varying in oncogenic potential, the procedure might have useful application in elucidating HSV oncogenesis.

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