No abstract

Cell cultures of simian and human origin infected with two strains each of herpesvirus saimiri (HVS) and herpesvirus ateles (HVA) were compared for production of infectious virus and early and late antigens (EA, LA) in the presence and absence of the tumor promoting agent 12-0-tetradecanoylphorbol-13-acetate (TPA). Second, Epstein-Barr virus (EBV) infected human and simian lymphoblastoid cell lines of high and low cell passages were also compared for enhanced production of early antigen and virus capsid antigen (VCA) using two concentrations of TPA, owl monkey kidney (OMK) and squirrel monkey lung. Cell cultures infected with HVS and HVA with 20 ng/ml of TPA exhibited higher percentages of early and late antigen producing cells and contained 1.0-1.6 logs more virus. Such cells also had earlier cytopathic effects, larger plaques, and a 3-fold increase in the number of plaques. The TPA also enhanced HVS-EA, and LA both in OMK and human skin fibroblast (HSF) cells. The enhancement of EA was approximately 17.0% more in OMK cells and 4.0% more in HSF. The HVS-LA was 22% higher in OMK and 6.0% higher in HSF cells. Pretreatment of OMK cells with TPA prior to HVS or HVA infection showed only a 0.5-0.7 log enhancement of virus. A dose-response of TPA in P,HRl cells showed that both 20 and 40 ng/ml doses were able to enhance EBV, EA, and VCA significantly with peak enhancement at 40 ng/ml. Higher doses of TPA (60 and 100 ng/ml) resulted in considerable cell death and reduction in antigen production. TPA (20 and 40 ng/ml) stimulated both VCA and EA antigen production in P3HR1 and B95-8 cells, with lesser effects on other human and simian lymphoblastoid cells. However, the stimulation of EA and VCA with these doses of TPA varied for each cell line. Moreover, TPA-treated P,HRl, B95-8, and 407-1 cells on the average also produced 1.0-1.5 logs more virus than the untreated cells. The higher percentage of EA-VCA production from P3HRl and B95-8 cells and lower EA-VCA from other human and simian cells suggests that such virus-cell interaction may be influenced by in vitro passages.

A combination of technical advances (most notably heterologous cell fusion, high resolution G-banding, and molecular cloning) has contributed to an accelerated advance in genetic analysis in mammals. The present human genetic map contains over 400 gene assignments and the map is growing rapidly as each ncw molecular clone or immunological reagent is developed. In our laboratory, we have devclopcd a panel of rodent x human somatic cell hybrids that have been utilized in chromosomc assignment of several classes of genes including oncogenes @us, ruj) and endogcnous human retroviral sequences (ERM, 2, etc). Using similar techniques, a biochemical gcnetic map of the domestic cat has been derived. The cat has 19 chromosome pairs and, to date, 40 genes have been mapped to 16 linkage or syntenic groups. Comparison of linkage relationships between homologous enzymes has revealed a striking conversation of chromosomal linkage association between cat and man. A comparison of syntenically homologous, highly extended high resoultion G-banded chromosomes between the two mammalian families revealed that 20-25%, by length, of the human karyotype can be precisely aligned (chromomere to chromomere) between cats and man despite the evolutionary divergence of the species nearly 80 million years ago.Moderately repetitive families of retrovirus-related DNAs exist within the feline and the human genomes. We have isolated molecular clones of several members of the feline RD-114 retrovirus family from a genomic library of normal cat cellular DNA. The endogenous sequences analyzed were similar to each other in that they were colinear with RD-114 proviral DNA, were bounded by long terminal redundancies, and conserved many restriction sites in the gag and pol regions. Several sequences were apparently deleted, relative to the previously characterized inducible RD-114 gcnome. The env regions of a number of endogenous RD-114 sequences examined were suhstantially deleted or diverged; a subset of these sequences contained information at the position of the env region that was not homologous to inducible RD-114. The RD-114 virogenes were dispersed to several cat *Based on a lecture delivered in September 1983, at the International Conference in Biochemical and Developmental Genetics, Kos, Greece.