The cellular basis of cardiac allograft rejection: VIII. Mechanisms underlying delayed allograft rejection in PVG C6-deficient rats

Background. The delayed allograft rejection in C6-deficient PVG C6^- rats compared with normal PVG rats has been attributed to the lack of alloantibody activation of the membrane attack complex of complement. As T cells alone have been shown to effect graft rejection, we examined T-cell responses in PVG C6^- rats. Methods. The cellular infiltrate and its mRNA for cytokines and effector molecules in DA heart allografts to PVG and PVG C6^- rats was compared by immunoperoxidase staining and semiquantitative reverse transcriptase polymerase chain reaction. The ability of pure populations oft cells or alloantibody to mediate DA heart graft rejection in irradiated (750 tads) PVG and PVG C6^- rats was also compared. Results. The median rejection time of DA heart allografts was 8 days in PVG rats and 17.5 days in PVG C6^- . PVG C6^- rats sensitized to DA by two skin grafts rejected DA heart grafts in 5-6 days. CD3⁺, CD4⁺, CD8⁺, interleukin-2 receptor-positive T cell, macrophage, and natural killer cell infiltration, as well as class II major histocompatibility complex and intercellular adhesion molecule-1 up- regulation, in grafts was similar in naive PVG and PVG C6^- rats. mRNA for T helper 1 cytokine interleukin-2, interferon-γ, tumor necrosis factor-β, macrophage molecules tumor necrosis factor-α, and inducible nitric oxide synthase, as well as cytotoxic T-cell effector molecules perforin and granzyme A and B, were found to be the same in the grafts from both naive PVG and naive PVG C6^- rats. Thus, there appeared to be no difference in the T- cell effector response between the PVG and PVG C6^- groups. There were higher alloantibody titers in PVG C6^- rats than in PVG hosts. Irradiation ablated rejection and alloantibody responses and reconstitution with naive T cells alone restored rejection in both PVG and PVG C6^- rats. Irradiated rats given serum from PVG rats that had rejected DA grafts did not effect rejection of DA grafts even if given naive T cells. Sensitized T cells restored second set. Conclusions. PVG C6^- rats have normal T-cell responses and can mediate allograft rejection in the absence of alloantibody. The failure of PVG C6^- to reject allografts rapidly may be a result of the poor clearance of alloantisera leading to enhancement of graft survival rather than a critical role for complement and membrane attack complex in acute rejection.