Improvement of Herpesvirus saimiri T cell immortalization procedure to generate multiple CD4+ T-cell clones from peripheral blood lymphocytes of AIDS patients.

Herpesvirus saimiri (HVS) can infect and immortalize human T lymphocytes of both CD4- and CD8-positive phenotypes. We have previously shown that infection of peripheral blood lymphocytes (PBL) from AIDS patients with HVS predominantly yielded immortalized CD8-positive T cell clones. Here we show that CD4-positive T cells from AIDS patients can be efficiently immortalized by HVS if patient PBL are enriched for CD4-positive T cell subpopulation prior to HVS infection. Such cells can be cloned and maintained in culture for prolonged times, and they exhibit activated T cell phenotype of Th1 class and are susceptible to HIV-1 infection. Several immortalized T cell clones obtained from one out of three AIDS patients tested here were HIV-1 positive and produced infectious virus. This approach permits efficient generation of multiple CD4-positive T cell clones from AIDS patients for functional and virological studies.