The ability of cultured lymphoblasts to stimulate autologous lymphocyte transformation in "one-way" mixed leukocyte culture has been studied. Intact, cultured lymphoblasts required physical contact with responding lymphocytes to induce transformation. In quantitative terms, lymphocytes incorporate as much thymidine when mixed with irradiated cultured lymphoblasts as they do in response to phytohemagglutinin. The stimulation of lymphocyte transformation by allogeneic cultured lymphoblasts did not parallel the stimulation of lymphocyte transformation by leukocytes from the donor of the lymphoblast culture. The stimulatory determinants on the cultured lymphoblast are unaffected by neuraminidase but destroyed by trypsin. The trypsin-treated cultured lymphoblasts regain their capacity to stimulate autologous lymphocyte transformation within 48 hr in culture. Cultured lymphoblasts possess concanavalin A binding sites. Concanavalin A inhibits the capacity of cultured lymphoblasts to stimulate autologous lymphocyte transformation. The relevance of these findings to EB virus infection of cultured lymphoblasts and to immune surveillance is discussed.