Failure to develop antibodies to nonself A and B blood group antigens is well described after infant ABO-incompatible heart transplantation and suggests that exposure to incompatible ABO antigens early in life may lead to tolerance rather than immunogenicity. If this finding is also true following ABO-incompatible cryopreserved homograft implantation, then such patients who require transplantation may be able to accept certain ABO-incompatible organs. In this study, we measured anti-A and -B antibody titers (isohemagglutinins) and allosensitization to human leukocyte antigens (HLA) in 21 patients after homograft placement (12 of whom were <1 year of age at initial homograft exposure) in childhood. We also examined homograft explant specimens for endothelial preservation and expression of HLA and A and B blood group antigens. We observed no differences in isohemagglutinins between patients who received ABO-incompatible versus ABO-compatible homografts. Allosensitization to HLA was present in 88% of patients (9 of 9 ABO-incompatible recipients and 5 of 7 ABO-compatible recipients). In 7 homograft explant specimens (median implant duration 10.1 years), the vasa vasorum endothelium was intact with ABO blood group antigen expression on 3 of 5 non-O homografts. These data suggest that tolerance to incompatible A and B blood group antigens does not occur following placement of ABO-incompatible homografts in childhood.