Tregs Expanded In Vivo by TNFRSF25 Agonists Promote Cardiac Allograft Survival

Background. Thymic-derived Foxp3(+)CD4(+) regulatory T cells (Tregs)-also called natural Tregs-are critical for the induction and maintenance of transplantation tolerance. Using an agonistic tumor necrosis factor-receptor super family (TNFRSF) 25 antibody, clone 4C12, we showed that TNFRSF25 is a powerful regulator of Treg proliferation-mediating expansion of natural Tregs in vivo. In the present study, we investigate the role of Tregs expanded in vivo by TNFRSF25 on cardiac allograft survival in a mouse model of fully major histocompatibility complex-mismatched ectopic heart transplants. Methods. C57BL/6 mice were treated with 20 mu g of TNFRSF25 agonist 4C12 4 days before heterotopic allogeneic heart transplantation. The survival of the graft was monitored daily by abdominal palpation until the cessation of cardiac contraction. The severity of immune rejection was evaluated by histopathology. Infiltration of inflammatory cells and Tregs into the graft were characterized by flow cytometry. The expression of cytokines and other regulatory proteins was measured by quantitative real-time polymerase chain reaction. Results. Treatment with 4C12 resulted in expansion of Tregs to 30%-35% of CD4(+) cells and was associated with a significant prolongation of median graft survival from 8 days to 17 days (P=0.0049). On day 7 after transplantation, the time point when controls reject the graft, the transplants of 4C12-treated animals beat strongly and showed increased accumulation of Foxp3(+) Tregs within the graft and decreased infiltration of inflammatory cells. Conclusions. TNFRSF25 agonists expand Tregs in vivo and delay allograft rejection.