Ex Vivo Expanded Human Non-Cytotoxic CD8+CD45RClow/− Tregs Efficiently Delay Skin Graft Rejection and GVHD in Humanized Mice

Abstract

Both CD4⁺ and CD8⁺ Tregs play a critical role in the control of immune responses and immune tolerance; however, our understanding of CD8⁺ Tregs is limited while they are particularly promising for therapeutic application. We report here existence of highly suppressive human CD8⁺CD45RC^low/- Tregs expressing Foxp3 and producing IFNγ, IL-10, IL-34, and TGFβ to mediate their suppressive activity. We demonstrate that total CD8⁺CD45RC^low/- Tregs can be efficiently expanded in the presence of anti-CD3/28 mAbs, high-dose IL-2 and IL-15 and that such expanded Tregs efficiently delay GVHD and human skin transplantation rejection in immune humanized mice. Robustly expanded CD8⁺ Tregs displayed a specific gene signature, upregulated cytokines and expansion in the presence of rapamycin greatly improved proliferation and suppression. We show that CD8⁺CD45RC^low/- Tregs are equivalent to canonical CD4⁺CD25^highCD127^low/- Tregs for suppression of allogeneic immune responses <i>in vitro</i>. Altogether, our results open new perspectives to tolerogenic strategies in human solid organ transplantation and GVHD.