Recent studies indicate that Fas and Fas ligand are involved in apoptosis of T-cells in HIV-infected patients. We have demonstrated that calcium/calmodulin is involved in Fas-mediated apoptosis in human T-cell lines transfected with HIV recombinant cDNA. In the present study, we examined spontaneous apoptosis of T-cells in vitro in peripheral blood obtained from 11 patients with AIDS and 8 HIV-seronegative normal donors and the effect of the calmodulin antagonists, trifluoperazine (TFP) or tamoxifen (TMX), on apoptosis. The results show that: (1) levels of spontaneous apoptosis were higher in PBMCs obtained from patients with AIDS than HIV-negative normal controls and the levels of apoptosis correlated with the severity of disease. (2) The accelerated apoptosis occurred predominantly in CD4+ cells in patients with AIDS. (3) Calmodulin antagonists inhibited the spontaneous apoptosis of CD4+ T-cells from patients with AIDS, which resulted in an increase in the ratio of CD4+ to CD8+ T-cells. (4) The inhibitory effect of calmodulin antagonists on apoptosis was more significant in patients with advanced disease (CDC category C) compared to less severe disease (CDC category B). These results indicate that calmodulin antagonists inhibit HIV-associated apoptosis of CD4+ T-cells, and imply that the calcium/calmodulin play important roles in mediating apoptosis of CD4+ T-cells induced by HIV infection.