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Studies of simian virus 40-infected human cell lines
Abstract
Simian virus 40 (SV40) infections of semipermissive human cells and permissive monkey cells were compared. Viral specific events were examined on per cell basis using immunocytochemistry to detect viral antigen-producing cells and in situ hybridization to identify viral DNA-replicating cells. The SV40 infection aborted before T-antigen expression in many cells of each of the human cell lines examined. This was further investigated by in situ hybridizations to probe for early SV40 message. The results showed that the same number of cells synthesize early message as produce T antigen indicating that the block to T-antigen expression is prior to transcription. In all but one of the human cell lines studied, most of the T-antigen-producing cells replicated viral DNA. However, in the A172 line of human glial cells only a small percentage of the T-antigen-expressing cells replicated viral DNA. Different structural and functional classes of T antigen have been characterized using anti-T monoclonal antibodies. Therefore, infected A172 cells were examined with a panel of ten anti-T monoclonal antibodies to determine if viral DNA replication might correlate with the expression of a particular epitope of T antigen. One anti-T monoclonal, PAb 100, did specifically recognize that subset of A172 cells which replicated SV40 DNA. The percentage of PAb 100-reactive A172 cells was dramatically increased by the DNA synthesis inhibitors hydroxyurea and aphidicolin. Removal of the hydroxyurea was followed by an increase in the percentages of cells replicating viral DNA corresponding to the increased percentage reactive with PAb 100. The pattern of SV40 infection in A172 cells was not altered by infection with viable viral mutants containing lesions in the small t protein, the agnoprotein, or the enhancer region. Additional studies on carrier systems of SV40-infected A172 and rhesus cells are also described.
Subject Area
Microbiology|Cellular biology
Recommended Citation
Deminie, Carol Ann, "Studies of simian virus 40-infected human cell lines" (1991). Doctoral Dissertations Available from Proquest. AAI9120869.
https://scholarworks.umass.edu/dissertations/AAI9120869