Using two-color fluorescence flow cytometry, we were able to detect the presence of small numbers of T4+T8+ cells (about 3%) in freshly isolated peripheral T cell populations derived from normal healthy donors. Coexpression of T4 and T8 was predominantly found on large blastlike cells and appeared to be related to activation. Stimulation of peripheral T cells with concanavalin A (Con A) for 5 days resulted in the generation of up to 60% of T4+T8+ cells. Coexpression was accompanied by a twofold increase in the number of T8 antigenic sites per cell. The T4+T8+ cells in lectin-stimulated cultures expressed high levels of the activation antigens T9, T10, and the IL-2 receptor but lacked T6, an antigen found on a majority of stage II thymocytes. Coexpression of T4 and T8 appeared to be a transitory process, because prolonged culture of T cells in the absence of lectin resulted in the loss of the T4+T8+ phenotype. Our data suggest that T cell activation in peripheral blood results in the generation of a T4+T8+ cell population which is distinct from previously described thymic and peripheral blood cells. Because T4 and T8 molecules may interact directly with MHC antigens, coexpression of these molecules may have an important role in immune function.