CD25+CD4+ regulatory T cells prevent graft rejection:: CTLA-4- and IL-10-dependent immunoregulation of alloresponses

Specific and selective immunological unresponsiveness to donor alloantigens can be induced in vivo. We have shown previously that CD25(+)CD4(+) T cells from mice exhibiting long-term operational tolerance to donor alloantigens can regulate rejection of allogeneic skin grafts mediated by CD45RB(high)CD4(+) T cells. In this study, we wished to determine whether donor-specific regulatory cells can be generated during the induction phase of unresponsiveness, i.e., before transplantation. We provide evidence that pretreatment with anti-CD4 Ab plus a donor-specific transfusion generates donor-specific regulatory CD25(+)CD4(+) T cells that can suppress rejection of skin grafts mediated by naive CD45RB(high)CD4(+) T cells. Regulatory cells were contained only in the CD25(+) fraction, as equivalent numbers of CD25(-)CD4(+) T cells were unable to regulate rejection. This pretreatment strategy led to increased expression of CD122 by the CD25(+)CD4(+) T cells. Blockade of both the IL-10 and CTLA-4 pathways abrogated immunoregulation mediated by CD25(+) T cells, suggesting that IL-10 and CTLA-4 are required for the functional activity of this population of immunoregulatory T cells. In clinical transplantation, the generation of regulatory T cells that could provide dynamic control of rejection responses is a possible route to permanent graft survival without the need for long-term immunosuppression.