Unstable regulatory T cells, enriched for naïve and Nrp1^negcells, are purged after fate challenge

Abstract

Regulatory T cells (T_regs) are indispensable for the control of immune homeostasis and have clinical potential as a cell therapy for treating autoimmunity. T_regs can lose expression of the lineage-defining Foxp3 transcription factor and acquire effector T cell (T_eff) characteristics, a process referred to as T_reg plasticity. The extent and reversibility of such plasticity during immune responses remain unknown. Here, using a murine genetic fate-mapping system, we show that T_reg stability is maintained even during exposure to a complex microbial/antigenic environment. Furthermore, we demonstrate that the observed plasticity of T_regs after adoptive transfer into a lymphopenic environment is a property limited to only a subset of the T_reg population, with the nonconverting majority of T_regs being resistant to plasticity upon secondary stability challenge. The unstable T_reg fraction is a complex mixture of phenotypically distinct T_regs, enriched for naïve and neuropilin-1-negative T_regs, and includes peripherally induced T_regs and recent thymic emigrant T_regs These results suggest that a "purging" process can be used to purify stable T_regs that are capable of robust fate retention, with potential implications for improving cell transfer therapy.