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The portal venous (p.v.) administration of foreign cells induces donor‐specific tolerance. Recently, we have demonstrated that the p.v. administration of donor cells elicits donor‐specific tolerance across major histocompatibility complex barriers. In the present study, utilizing the intrahepatic tolerance‐inducing system, we have established a new method for organ transplantation using both busulfan ([Bu] to provide a sufficient “space” for the donor hematopoietic cells to expand in the recipient) and neuraminidase ([Neu] to enhance the trapping of i.v.‐injected cells in the liver).  Radiolabeled bone marrow cells (BMCs) were found to exclusively accumulate in the livers of the recipients as a result of the Neu treatment. Furthermore, hematopoietic progenitors (forming hematopoietic foci) in the accumulated BMCs were retained in the recipient livers for at least 18 days.  C57BL/6 (B6) mice that had been transplanted with skins of BALB/c mice immediately after the injection of BALB/c BMCs showed a 90% skin graft survival rate over 400 days as a result of using the combination of injecting 50 mg/kg Bu into the B6 mice and treatment of the BALB/c BMCs with 0.25 U/ml Neu (50 Bu + 0.25 Neu). However, the survival rate significantly decreased when either the Bu or Neu treatment was omitted. In tolerant recipients, microchimerism was observed in the various hematolymphoid organs. T cells collected from the tolerant recipients suppressed proliferative responses to the donor‐alloantigens but enhanced the production of Th2 and Th3 cytokines.  These findings suggest that the enhanced retention of donor BMCs in the recipient livers as a result of the Bu and Neu treatments efficiently induces tolerance induction. Therefore, this “single‐day protocol” would be of great advantage for human organ transplantation.

A New Method for Tolerance Induction: Busulfan Administration Followed by Intravenous Injection of Neuraminidase‐Treated Donor Bone Marrow