C5aR1 regulates migration of suppressive myeloid cells required for costimulatory blockade-induced murine allograft survival

Costimulatory blockade-induced murine cardiac allograft survival requires intragraft accumulation of CD11b ⁺ Ly6C ^lo Ly6G ⁻ regulatory myeloid cells (Mregs) that expand regulatory T cells (Tregs) and suppress effector T cells (Teffs). We previously showed that C5a receptor (C5aR1) signaling on T cells activates Teffs and inhibits Tregs, but whether and/or how C5aR1 affects Mregs required for transplant survival is unknown. Although BALB/c hearts survived >60 days in anti-CD154 (MR1)-treated or cytotoxic T-lymphocyte associated protein 4 (CTLA4)-Ig–treated wild-type (WT) recipients, they were rejected at ~30 days in MR1-treated or CTLA4-Ig–treated recipients selectively deficient in C5aR1 restricted to myeloid cells (C5ar1 ^fl/fl xLysM-Cre). This accelerated rejection was associated with ~2-fold more donor-reactive T cells and ~40% less expansion of donor-reactive Tregs. Analysis of graft-infiltrating mononuclear cells on posttransplant day 6 revealed fewer Ly6C ^lo monocytes in C5ar1 ^fl/fl xLysM-Cre recipients. Expression profiling of intragraft Ly6C ^lo monocytes showed that C5aR1 deficiency downregulated genes related to migration/locomotion without changes in genes associated with suppressive function. Cotransfer of C5ar1 ^fl/fl and C5ar1 ^fl/fl xLysM-Cre myeloid cells into MR1-treated allograft recipients resulted in less accumulation of C5ar1 ^{− / −} cells within the allografts, and in vitro assays confirmed that Ly6C ^hi myeloid cells migrate to C5a/C5aR1-initiated signals. Together, our results newly link myeloid cell–expressed C5aR1 to intragraft accumulation of myeloid cells required for prolongation of heart transplant survival induced by costimulatory blockade.