Apoptosis can be induced in the human leukemic T-cell line Jurkat when these cells are stimulated by several known T-cell activators. Among the stimuli that activate Jurkat cells, calcium ionophore A23187, phytohemagglutinin and cross-linked monoclonal antibody anti-CD3 are the best inducers of apoptosis. The activator of protein kinase C phorbol-12,13-dibutyrate causes a rapid but transient stimulation of DNA fragmentation and cell death, and soluble monoclonal antibody anti-CD3 has no effect. Furthermore, apoptosis is delayed when Jurkat cells are stimulated simultaneously with calcium ionophore and phorbol ester. We have analysed the mechanism underlying the induction of apoptosis by these agents and found that down-regulation of c-myc expression by pretreatment with the DNA binding antibiotic mithramycin, an inhibitor of c-myc transcription, does not prevent the activation of the apoptotic process. In the light of these results we can conclude that in these leukemic T-cells, activation of apoptosis may occur by a mechanism independent of c-myc expression.