The replication of herpes simplex virus (HSV) in cultures of human T lymphocytes was investigated. Virus replication occurred only in lymphocyte cultures prestimulated with mitogen. At least one of the factors responsible for this phenomenon is adsorption of the virus to the cells, which was 5 times less efficient in nonstimulated cells than in stimulated cultures. Growth of virus in infected cultures was restricted to lymphocytes of T origin. This was shown by successful infection of highly purified T-cell cultures and by virus replication in four continuous T-cell lines. No virus production was obtained in cultures of lymphocytes of other than T-cell origin. Virus titers in T-lymphocyte cultures reached 107 PFU/ml, which was a four logio step increase over the input dose. Lymphocytes infected with a syn strain of HSV, HSV-ANG, induced polykaryocyte formation in the cultures and therefore generated a CPE visible by microscopic examination. Infection with syn+ strains of HSV (i.e., strains which do not cause cell fusion) did not lead to such phenomena. Not more than 1% of infected T cells was replicating HSV, as revealed by infectious center assays. In contrast, about 5% of these cells showed positive immunofluorescence with anti-HSV antibodies, indicating the presence of cells which express viral protein but do not actively produce infectious virus.
Herpes simplex virus (HSV) replicated in mitogen-stimulated human T cells. Virus replication was obtained in highly enriched mitogen-stimulated T cells of the OKT 3+, OKT 4+, or OKT 8+ subtype, in stimulated B cells, and in macrophages precultured for 7 days. In contrast, no virus replication was obtained in unstimulated T or B cells, in macrophages grown in culture for 1 day, in Null/NK cells, or in granulocytes. Infectious center assays revealed that below 1% of the infected T cell subpopulations supported virus replication, whereas up to 42% of infected B cells and 80% of macrophages cultured for 1 wk were able to replicate HSV. By indirect double immunofluorescence studies, complement-mediated mass cytolysis, and positive selection experiments, it was shown that only T cells expressing Ia antigen actively replicated the virus. T cells activated in the mixed lymphocyte culture and with UV-inactivated HSV were also susceptible to HSV infection. Several human leukocyte cell lines were tested for their ability to support virus replication and were tested for a correlation with the expression of Ia antigen. Only cell lines expressing Ia antigen on more than about 5% of the total population produced new progeny virus. Ia-expressing T cells that spontaneously replicated HSV without any mitogenic prestimulation were found to occur in variable numbers in human cord blood. It is suggested, that such T cells, permissive for HSV replication, might contribute to an outspread of viral infection in vivo.
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