PREVENTION AND RESTORATION OF SECOND-SET LIVER ALLOGRAFT REJECTION IN PRESENSITIZED MICE

Abstract

The aim was to determine whether sublethal donor total body irradiation (TBI) might be as effective as lethal TBI in preventing mouse second-set liver allograft rejection, and to evaluate the role of passenger leukocytes, donor major histocompatibility complex (MHC) antigens, and host effector mechanisms in the response to livers from sublethally irradiated donors. B10 (H2b) donors received various doses of TBI at different times before their livers were transplanted orthotopically into normal or donor skin-presensitized C3H (H2k) recipients. The influence of irradiation on graft non-parenchymal cells (NPC) was determined by monoclonal antibody staining, and flow cytometric analysis. Hematopoietic cells within the grafts were reconstituted by intravenous infusion of syngeneic or third-party bone marrow cells. Allograft survival was determined in recipients that received no treatment, or that were given spleen cells from either normal B10 donors, or MHC class I - or class II-deficient mice syngeneic with the donors. Cytotoxic activity of graft-infiltrating cells and host spleen cells, and complement-dependent cytotoxic alloantibody titers were determined by isotype release assays. The protective effect of donor TBI was observed both at lethal (9.5 Gy) and sublethal doses (5 and 3 Gy; graft median survival time: >100 days). Extended delay in liver transplantation, allowing hematopoietic recovery and graft reconstitution eliminated the effect. Liver NPC were reduced about 80% within 24 hr of 3 Gy TBI, with a selective reduction in the incidence of B cells. The NPC-depleted livers underwent accelerated rejection when donor (but not third-party) spleen cells (5 x 10(7) were administered systemically to the recipient immediately after graft revascularization. Spleen cells from MHC class I-deficient (but not MHC class II-deficient) mice failed to fully restore accelerated rejection of TBI liver grafts. Freshly isolated graft NPC, or spleen cells from TBI liver recipients, harvested 4 days after transplantation, exhibited lower, donor-specific cytotoxic activity than cells from mice given normal livers. Recipients of TBI livers also showed much lower serum complement-dependent cytotoxic alloantibody titers. By substantially depleting "passenger leukocytes," sublethal donor TBI undermines anti-donor cell-mediated and humoral immune reactivity and inhibits second-set liver allograft rejection in presensitized recipients. The interval between irradiation and transplantation is important in conferring resistance to rejection. Expression of MHC class I on donor leukocyte infusions is important for overcoming resistance to second-set rejection induced by donor irradiation.