Therapeutic Efficacy of Polyclonal Tregs Does Not Require Rapamycin in a Low-Dose Irradiation Bone Marrow Transplantation Model

Background. Mixed chimerism is an effective strategy for the induction of transplantation tolerance but the toxicity of recipient conditioning makes current bone marrow (BM) transplantation (BMT) protocols unsuitable for widespread clinical application. Therapies promoting BM engraftment under minimal conditioning would facilitate translation of this concepttotheclinic.Recently,wehaveshownthatregulatoryTcell(Treg)therapyhaspotentengraftment-enhancingeffects in an irradiation-free noncytotoxic BMT protocol, but only if it is combined with rapamycin treatment. Methods. Here, we investigated whether polyclonal Treg therapy is effective in promoting chimerism and tolerance in anotherwiseunsuccessfulBMTprotocolusinglow-dosetotalbodyirradiation(1Gy)andcostimulationblockadeand determined whether Tregs do so on their own without rapamycin. Results. The application of polyclonal FoxP3-transduced recipient Tregs led to durable multilineage chimerism and donor-specific skin graft tolerance whereas recipients receiving costimulation blockade alone or green flourescent protein (GFP)-transduced cells failed to develop chimerism. Infused Tregs had a limited life span as indicated by polymerasechainreactionanalysisbutrathercontributetodenovoinductionofsubsequentTreggenerations.Deletion of donor-reactive T cells was observed but progressed more slowly over time compared with recipients of a nonmyeloablative BMT protocol using 3 Gy total body irradiation. Conclusions. In conclusion, Treg therapy promotes BM engraftment on its own in a low-dose irradiation BMT protocol, leading to chimerism and tolerance maintained through deletional and nondeletional mechanisms.