Unrelated donor (URD) bone marrow transplantation (BMT) is associated with more frequent and more therapy-resistant graft-versus-host disease (GVHD). We tested an in vivo immunotoxin with direct cytolytic potency against CD5-expressing T lymphocytes (Xomazyme-CD5) for GVHD prophylaxis after URD BMT. The immunotoxin was given in vivo (0.1 mg/kg/day) for 3 weeks following transplantation in combination with methotrexate+prednisone (MXP; n = 16) or methotrexate+cyclosporine (MCX; n = 6). The 22 patients (10 phenotypically matched with their donors and 12 partially matched) received unmanipulated marrow. MXP was well tolerated, while MCX led to unacceptable nephrotoxicity, weight gain and edema. Four patients died of early complications. Thirteen of 17 evaluable patients achieved myeloid engraftment by 17-40 days (median 24 days). Acute GVHD developed in 9 of 15 evaluable patients (5 grade III/IV). Six of 8 evaluable patients developed chronic GVHD. Four patients survive 1.1-2 years after BMT. Although this immunotoxin has previously shown potency in prophylaxis of murine GVHD and therapy of human GVHD, in this trial inadequate immunosuppressive potency of the immunotoxin combinations was associated with unacceptable clinical toxicity. Aggressive immunoprophylaxis against GVHD is required to improve the success of URD BMT.