Complex Ca2+ flux inhibition as primary mechanism of staurosporine-induced impairment of T cell activation.

The inhibitory effect of the highly effective drug staurosporine on the early activation signal Ca2+ flux was investigated via multiparameter flow cytometry in human peripheral blood T lymphocytes. Staurosporine has been reported to be a specific inhibitor of protein kinase C. However, we show that it inhibits the Ca2+ influx in anti-CD3 and phytohemagglutinin-stimulated human CD4+ and CD8+ lymphocytes at concentrations between 1.0 and 10.0 ng/ml. Staurosporine decreases the number of Ca2+-positive CD4+ and CD8+ lymphocytes as well as the Ca2+ influx per cell; the drug also delays the time of the maximum response to polyclonal stimulation. In addition, we demonstrate that staurosporine affects the primary Ca2+ response via inhibition of the release of the membrane-bound Ca2+ from the endoplasmic reticulum in CD4+ and CD8+ lymphocytes. Binding studies of the anti-CD3 antibody to T lymphocytes indicate normal binding capacities in the presence of staurosporine. With respect to the classical scheme of T cell activation via phospholipase C, our data suggest that staurosporine may inhibit T cell activation primarily by its effect on the early Ca2+ flux signal.