Efficient and selective prevention of GVHD by antigen-specific induced Tregs via linked-suppression in mice

Naturally occurring regulatory T cells (nTregs) suppress the development of graft-versus-host disease (GVHD) and may spare graft-versus-leukemia (GVL) effect. Because nTreg is a rare population in a healthy individual, the limited source and the non-selective suppression are major hurdles towards the application of nTregs in the control of clinical GVHD after allogeneic hematopoietic cell transplantation (HCT). An alternative approach is to generate induced Tregs (iTregs) from naïve CD4 precursors, but the effectiveness of iTregs in the control of GVHD is highly controversial and requires further investigation. The other critical but unsolved issue in Treg therapy is how to achieve antigen (Ag)-specific tolerance that distinguishes GVHD and GVL effects. To address the important issues on the effectiveness of iTregs and Ag-specificity of Tregs, we generated Ag-specific iTregs and tested their potential in the prevention of GVHD in a pre-clinical bone marrow transplantation (BMT) model. CD4(+)CD25(+)Foxp3(+) iTregs generated from OT-II TCR transgenic T cells specific for OVA target Ag efficiently prevented GVHD induced by polyclonal T effector cells (Teffs) only in the allogeneic recipients that express OVA protein but not in OVA(-) recipients. The efficacy of these Ag-specific iTregs was significantly higher than polyclonal iTregs. As controls, OT-II CD4(+)Foxp3(-) cells had no effect on GVHD development in OVA(-) recipients and exacerbated GVHD in OVA(+) recipients when transplanted together with polyclonal Teffs. Because the iTregs recognize OVA whereas Teffs recognize alloAg bm12, our data reveal for the first time, to our knowledge, that Tregs prevent GVHD through a linked suppression. Mechanistically, OT-II iTregs expanded extensively, and significantly suppressed expansion and infiltration of Teffs in OVA(+) but not in OVA(-) recipients. These results demonstrate that Ag-specific iTregs can prevent GVHD efficiently and selectively, providing a proof of principle that Ag-specific iTregs may represent a promising cell therapy for their specificity and higher efficacy in allogeneic HCT.