Author:
Steffen M,Krieg P,Pernfuss M,Sauer E,Eisinger V,Sauer G
Abstract
The stumptailed macaque papovavirus strain HD was discovered in a persistently infected cell line of primate origin designated Vero 76 (K. Bosslet and G. Sauer, J. Virol. 25:596--607, 1978; W. Waldeck and G. Sauer, Nature [London] 269:171--173, 1977). In clonal derivatives of Vero 76 cells a minor and variable proportion of cells is engaged in the productive synthesis of the HD virus strain. A combination of immunofluorescence using simian virus 40 polyoma subgroup-specific antiserum and in situ hybridization with HD complementary RNA revealed that only those cells which harbor discernible amounts of HD DNA also contain the subgroup-specific antigen. Treatment with arabinofuranosylcytosine caused irreversible disappearance of the antigen, whereas actinomycin D, in contrast, reversibly inhibited both HD DNA replication and synthesis of the subgroup-specific antigen. The proportion of HD DNA and subgroup-specific antigen-synthesizing cells in Vero 76 clonal lines could be either decreased or increased by the mode of passaging of the cell cultures. When cell cultures were split every 3 to 7 days at a 1:4 ratio, the amount of HD DNA sequences as revealed by DNA-DNA reassociation and by the Southern blotting technique fell below the level of detection after only a few passages. Furthermore, expression of the viral subgroup-specific antigen was no longer discernible. However, viral DNA persists in such latently infected cells, because a change in the splitting protocol to a 2-week passaging rhythm led to reinitiation of both viral DNA replication and expression of the subgroup-specific antigen. The HD DNA is perpetuated in a restricted state in latently infected cells in an episomal, unintegrated form as shown by Southern blot analysis. This finding complies with the fact that HD DNA-free subclones could be derived from persistently infected clonal Vero 76 cells. Such subclones have lost the viral genomes, probably owing to segregation during cell division.
Publisher
American Society for Microbiology
Subject
Virology,Insect Science,Immunology,Microbiology
Cited by
6 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献