A critical question facing the field of transplantation is how to control effector T cell (T_eff) activation while preserving regulatory T cell (T_reg) function. Standard calcineurin inhibitor–based strategies can partially control T_effs, but breakthrough activation still occurs, and these agents are antagonistic to T_reg function. Conversely, mechanistic target of rapamycin (mTOR) inhibition with sirolimus is more T_reg-compatible but is inadequate to fully control T_eff activation. In contrast, blockade of OX40L signaling has the capacity to partially control T_eff activation despite maintaining T_reg function. We used the nonhuman primate graft-versus-host disease (GVHD) model to probe the efficacy of combinatorial immunomodulation with sirolimus and the OX40L-blocking antibody KY1005. Our results demonstrate significant biologic activity of KY1005 alone (prolonging median GVHD-free survival from 8 to 19.5 days), as well as marked, synergistic control of GVHD with KY1005 + sirolimus (median survival time, >100 days; P < 0.01 compared to all other regimens), which was associated with potent control of both T_H/T_C1 (T helper cell 1/cytotoxic T cell 1) and T_H/T_C17 activation. Combined administration also maintained T_reg reconstitution [resulting in an enhanced T_reg/T_eff ratio (40% over baseline) in the KY1005/sirolimus cohort compared to a 2.9-fold decrease in the unprophylaxed GVHD cohort]. This unique immunologic signature resulted in transplant recipients that were able to control GVHD for the length of analysis and to down-regulate donor/recipient alloreactivity despite maintaining anti–third-party responses. These data indicate that combined OX40L blockade and sirolimus represents a promising strategy to induce immune balance after transplant and is an important candidate regimen for clinical translation.