A number of T cell surface antigens including CD45R0, CD58, CD11 alpha, CD29, CD44, and CD26 are present on differentiated T cells and identify T cell populations that respond to recall antigens. To further study the biochemical basis for this immune memory, we used an anti-CD26 mAb to identify the memory T cell population. We have previously shown that CD26+ T cells display an increased proliferative response to anti-CD3 and anti-CD2 mAbs and furthermore have significantly greater PMA-induced phosphorylation of the invariant gamma and delta chains of the T cell receptor (TCR)/CD3 complex when compared to CD26-T cells. This suggested that differential distribution of protein kinase C (pkC) in the CD26 subsets may be related to the reduced activation requirements observed in memory T cells upon recall antigen challenge. We now directly demonstrate that when peripheral blood T cells are sorted into CD26+ and CD26- T cells the majority of pkC activity can be recovered from the cytosol fraction of the memory (CD26+) T cell population. When activated through the TCR/CD3 pathway, the CD2 pathway, or directly by the phorbol ester, PMA, the memory (CD26+) T cells showed an increased proliferative response that was inhibited by the pkC inhibitor, staurosporine. When CD26+ T cells were cultured in the presence of PMA, which depletes pkC activity, CD26 antigen expression was down-regulated. PMA was also able to inhibit phytohemagglutinin (PHA)-induced expression of the CD26 antigen in CD26- T cells. These data demonstrate a relation between CD26 expression and pkC activity and suggest that enhanced pkC activity is associated with memory T cell function.