It has been considered before that human naive and memory/effector CD4+ T-cells cannot be subdivided solely according to the differential expression of CD45 isoforms. By the lack of expression of CD31 we have identified a subset of CD4+ CD45RA+ CD31- cells which show distinct features of antigen-experienced Th1 cells. Short term stimulation of highly purified human peripheral blood CD4+ T-cells with PMA/ionomycin, followed by the cytometric analysis of intracellular cytokines, showed that a minor subpopulation of CD4+ CD45RA+ CD45RO- cells is able to produce interferon-gamma (IFN-gamma) rapidly, a characteristic of antigen-experienced Th1 cells. Whereas among CD45RA+ CD4+ T-cells both CD31+ and CD31- subsets produce interleukin-2 (IL-2) upon PMA/ionomycin stimulation, only the CD31- subpopulation is able to produce IFN-gamma. Thus, our phenotypic and functional characterization of CD45RA+ CD45RO- Th cells shows that CD45RA+ CD45RO- cells do not represent a homogeneous population of antigen-unexperienced, naive T-cells. We speculate that a certain subset of human CD4+, CD45RO+ memory T-cells reverts to expression of the CD45RA isoform, and that this subset can be identified by the lack of CD31 expression.