Transformation of Rat Embryo Cells by Temperature-sensitive Mutants of Herpes Simplex Virus

Abstract

SUMMARYRat embryo cells have been transformed by temperature-sensitive mutants of herpes simplex type 2 and by wild-type virus of herpes simplex types I and 2. Transformation was scored using the morphological criterion of focus formation and the cell changes that lead to the final focus are described.Herpes specified antigens can be detected in at least 5o ~ of these transformed rat embryo cells by immunofluorescence studies on unfixed material.Transformation experiments using hamster embryo cells and HSV-2 strain 333 are also described. These transformed cells are shown to be tumourigenic in newborn hamsters. Tumour cells in turn, as a single cell suspension, produce fresh tumours in animals up to 2 weeks old with almost Ioo ~ efficiency. Tumour cells also continue to retain the herpes specific antigens.

Induction of a Latent Herpes Simplex Virus from a Rat Tumour Initiated by Herpes Simplex Virus-transformed Cells

“…Nor, indeed, has treatment of explanted tumour cells with IUdR, BUdR or TPA succeeded in induction of replicating virus. These results are similar to those reported by Macnab (1974) where it was not possible to isolate virus from HSV-2-transformed cell lines.…”

Herpes Simplex Virus and Human Cytomegalovirus: Their Role in Morphological Transformation and Genital Cancers

“…One result, previously interpreted as the continuing expression of HSV functions in transformed cells (Macnab, 1974(Macnab, , 1975(Macnab, , 1979Bfifiltjens & Macnab, 1981) remained to be explained. When cells were initially transformed they formed a focus, which was subcultured and the cells were tested for immunofluorescence.…”

“…Both tsK and tsD induce the large subunit of RR but its transforming function has been associated only with established cells Hayashi et al, 1985). Macnab (1974Macnab ( , 1975Macnab ( , 1979 and Cameron et al (1985) used primary cells, and therefore the large subunit of RR is not likely to be the transforming function.…”

“…When HSV ts mutants were used to transform cells (Macnab, 1974(Macnab, , 1975(Macnab, , 1976 and unpublished results) all, including HSV-1 tsK, a DNA-negative mutant which produces only the four functional IE polypeptides [Vmw110 (ICP0), Vmw68 (ICP22), Vmw63 (ICP27), Vmwl2 (ICP47)], a non-functional Vmw 175 (ICP4) and Vmw136 (ICP6) (Marsden et al, 1976;Preston, 1979) were very cytotoxic (Schek & Bachenheimer, 1985) in the primary or secondary rat embryo cells used. Cytotoxicity varied from experiment to experiment.…”

Localization of the Coding Region for a 35000 Dalton Polypeptide on the Genome of Herpes Simplex Virus Type 2

“…Inactivated herpes simplex virus type 2 (HSV-2) (Duff & Rapp, 1971) and DNA or cloned DNA fragments from HSV-2 (Wilkie et al, 1974;Galloway & McDougall, 1981 ;Jariwalla et al, 1980;Macnab, 1974) have been used for oncogenic transformation of rodent fibroblasts in culture. In a number of laboratories, the presence of HSV-specific antigens has been detected in transformed cells, including membrane glycoproteins and the non-structural proteins VPI43 and ICP10 (Camacho & Spear, 1978;Duff& Rapp, 1973;Lewis et al, 1982;Reed et al, 1975;Flannery et al, 1977;Jariwalla et al, 1980).…”