The combination of total lymphoid irradiation and anti-T-cell antibodies safely induces immune tolerance to combined hematopoietic cell and organ allografts in humans. Our mouse model required host natural killer T (NKT) cells to induce tolerance. Because NKT cells normally depend on signals from CD8⁺ dendritic cells (DCs) for their activation, we used the mouse model to test the hypothesis that, after lymphoid irradiation, host CD8⁺ DCs play a requisite role in tolerance induction through interactions with NKT cells. Selective deficiency of either CD8⁺ DCs or NKT cells abrogated chimerism and organ graft acceptance. After radiation, the CD8⁺ DCs increased expression of surface molecules required for NKT and apoptotic cell interactions and developed suppressive immune functions, including production of indoleamine 2,3-deoxygenase. Injection of naive mice with apoptotic spleen cells generated by irradiation led to DC changes similar to those induced by lymphoid radiation, suggesting that apoptotic body ingestion by CD8⁺ DCs initiates tolerance induction. Tolerogenic CD8⁺ DCs induced the development of tolerogenic NKT cells with a marked T helper 2 cell bias that, in turn, regulated the differentiation of the DCs and suppressed rejection of the transplants. Thus, reciprocal interactions between CD8⁺ DCs and invariant NKT cells are required for tolerance induction in this system that was translated into a successful clinical protocol.