It is now well established that allogeneic lymphocytes can mediate a potent graft-vs.-leukemia (GVL) reaction when administered to bone marrow transplant (BMT) recipients. The benefit of allogeneic lymphocyte transfusion is limited because many patients develop graft-vs.-host disease (GVHD) with prolonged pancytopenia, which sometimes proves fatal. The object of the present study was to determine the antileukemic potential and GVHD risk of in vivo-generated tumor-specific allogeneic T cells given shortly after BMT. BALB/C (H-2d) mice were inoculated with different cell doses (10(5) and 5 x 10(5)) of the A20 leukemia (BALB/C origin) 2 days prior to lethal total-body irradiation (TBI) and transplantation of allogeneic, major histocompatibility complex (MHC)-matched DBA marrow grafts (H-2d, minor difference to BALB/C). Donors of BM grafts and T cells were allogeneic, MHC-matched mice (DBA, H-2d, minor difference to BALB/C). Donor-type T cells were generated from mice immunized with irradiated A20 leukemia cells or nonmalignant BALB/C splenocytes and restimulated in vitro. Whereas no significant immunotherapeutic effect was seen in mice with high tumor burden (5 x 10(5)), allogeneic BMT in mice inoculated with 1 x 10(5) A20 cells resulted in a modest antileukemic effect. This survival rate remained unchanged when 10(6) T cells obtained from donors immunized with nonmalignant BALB/C derived cells were given posttransplantation. In contrast, a single injection of 10(6) T cells from leukemia-immunized donors led to potent GVL effects without mediating clinically overt GVHD. Our data provide evidence for the hypothesis that minimal residual disease can be eradicated without inducing GVHD by administering small amounts of specific allogeneic cytotoxic T cells after BMT.