Vpu increases susceptibility of human immunodeficiency virus type 1-infected cells to fas killing.

The importance of the Fas death pathway in human immunodeficiency virus (HIV) infection has been the subject of many studies. Missing from these studies is direct measurement of infected cell susceptibility to Fas-induced death. To address this question, we investigated whether T cells infected with HIV are more susceptible to Fas-induced death. We found that Fas cross-linking caused a decrease in the number of HIV-infected Jurkat T cells and CD4(+) peripheral blood leukocytes (PBLs). We confirmed this finding by demonstrating that there were more apoptotic infected than uninfected cells after Fas ligation. The increase in sensitivity of HIV-infected cells to Fas killing mapped to vpu, while nef, vif, vpr, and second exon of tat did not appear to contribute. Furthermore, expression of Vpu in Jurkat T cells rendered them more susceptible to Fas-induced death. These results show that HIV-infected cells are more sensitive to Fas-induced death and that the Vpu protein of HIV contributes to this sensitivity. The increased sensitivity of HIV-infected cells to Fas-induced death might help explain why these cells have such a short in vivo half-life.

Apoptosis in HIV-1 Infection.

We have studied apoptosis in lymph node and peripheral blood mononuclear cells (PBMCs) from HIV-infected children and adults and SIV-infected rhesus macaques. In lymph nodes, we found that apoptosis and productive infection occurred only rarely in the same cell. There was, however, a direct correlation between the numbers of apoptotic and productively-infected cells. In HIV-infected children, we found a direct correlation between disease severity and percentage CD4+ T cell apoptosis (p = 0.001). Both CD4+ and CD8+ T cell apoptosis were directly related to CD4+ T cell depletion (p = 0.006 and p = 0.01, respectively). In addition, we found a trend towards diminished CD4+ T cell apoptosis on anti-retroviral agents. These findings suggest that apoptosis of uninfected cells may be important in HIV pathogenesis and that measurement of apoptosis may be a useful marker of disease activity.

Apoptosis of CD4+ and CD8+ T cells isolated immediately ex vivo correlates with disease severity in human immunodeficiency virus type 1 infection.

Apoptosis of CD4+ and CD8+ T cells has been shown in peripheral blood mononuclear cells (PBMCs) from HIV-infected adults analyzed after overnight culture. Because cell death may be an artifact of in vitro culture, and because there is little information on apoptosis in pediatric HIV disease, we undertook a cross-sectional analysis of apoptosis in PBMCs analyzed immediately ex vivo in HIV-infected children and adults. PBMCs from 22 children, four adolescents, and nine adults and seronegative age-matched control subjects were stained for CD4 and CD8 surface markers. Apoptotic cells were detected in a newly characterized flow cytometric assay by diminished forward and increased side scatter. Children with the most advanced disease had 9.9% (SEM 1.8) apoptotic CD4+ T cells above control, significantly higher than in asymptomatic patients [0.4% (SEM 2.3)], those with mild disease [2.2% (SEM 1.83)], and those with moderate disease [2.5 (SEM 3.6)] (p = 0.015). The percentages of both CD4+ and CD8+ T cell apoptosis were directly related to CD4+ T cell depletion (R2 = 0.23; p = 0.006; n = 32 and R2 = 0.2; p = 0.012; n = 30, respectively). Patients who responded to antiretroviral therapy with the greatest increase in CD4+ T cell percentage had the least CD4+ T cell apoptosis (R2 = 0.15; p = 0.1; n = 19). These findings show that the rate or extent of T cell death by apoptosis percentage of T cell apoptosis is significantly increased in HIV-infected children. The observed correlation of both CD4+ and CD8+ T cell apoptosis with CD4+ T cell depletion suggests that apoptosis plays a role in HIV pathogenesis and may be a useful marker of disease activity.