We developed a new method of donor-specific blood transfusion (DST) for prospective kidney transplant recipients from a living related donor. Forty-four patients were transfused with 100 ml of one-haplotype matched kidney donor's whole blood at 1, 8, and 15 days after its storage. Ten of these patients received azathioprine (1.5 mg/kg per day) in addition to DST. After DST, three patients (6.8%) developed donor-specific lymphocytotoxic antibodies. The incidence of sensitization was not different between azathioprine-treated and untreated patients. Following DST, donor-specific mixed lymphocyte culture (MLC) was significantly suppressed without any accelerated (secondary type) response in early MLC. Subsequently, 24 patients received a kidney transplant from the donor. Graft survival rates were 96% and 90% at 12 and 24 months, respectively. Nine additional patients, seven from a two-haplotype matched sibling and two from a no-haplotype matched sibling also received DST. None of these patients became sensitized following DST, and all have functioning grafts for 6 to 18 months. This study indicates that 100 ml of stored whole blood DST three times at weekly intervals is a practical, less immunizing, and effective approach to enhance graft survival in recipients of a living related donor graft, DST produces donor-specific adaptive responses that might be conductive to successful graft outcome, and this protocol could be used in sibling donor-recipient pairs who do not share a haplotype.