Induction and detection of apoptosis in human periphery blood T-cells

Freshly isolated, human peripheral blood T (PBT) cells are resistant to induction of apoptosis. In this study, however, we have shown that although small numbers of monocytes (Mo) are required for PBT cells to proliferate optimally in response to mitogenic challenge, a relatively higher percentage of Mo results in a significant decrease in PHA-, but not ConA-induced T-cell proliferation. Interestingly, the decrease in T-cell proliferation correlated to an increase in apoptotic cell death. Moreover, ConA-induced PBT-cells underwent apoptosis in the presence of PHA-pretreated Mo, suggesting a key role of monocyte activation in this system. This apoptosis-promoting effect of activated Mo appeared to depend on contact or close proximity between Mo and PBT-cells, rather than via soluble mediators. Despite an increase in apoptosis by the presence of high numbers of Mo, PHA-stimulated PBT-cells released IL-2 at elevated levels proportional to the increasing numbers of Mo in cultures. They also expressed activation marker CD69 and the IL-2R-gamma chain on the cell surface at comparable or higher levels in the presence of high versus low numbers of Mo. These data suggest that PBT-cells can embark on a normal early phase of activation prior to undergoing apoptosis, thereby providing a model system to study how T-cells are committed to either proliferation or activation-induced apoptosis.