Pancreatic islet-specific engineered T_regs exhibit robust antigen-specific and bystander immune suppression in type 1...

Adoptive transfer of regulatory T cells (T_regs) is therapeutic in type 1 diabetes (T1D) mouse models. T_regs that are specific for pancreatic islets are more potent than polyclonal T_regs in preventing disease. However, the frequency of antigen-specific natural T_regs is extremely low, and ex vivo expansion may destabilize T_regs, leading to an effector phenotype. Here, we generated durable, antigen-specific engineered T_regs (EngT_regs) from primary human CD4⁺ T cells by combining FOXP3 homology-directed repair editing and lentiviral T cell receptor (TCR) delivery. Using TCRs derived from clonally expanded CD4⁺ T cells isolated from patients with T1D, we generated islet-specific EngT_regs that suppressed effector T cell (T_eff) proliferation and cytokine production. EngT_regs suppressed T_effs recognizing the same islet antigen in addition to bystander T_effs recognizing other islet antigens through production of soluble mediators and both direct and indirect mechanisms. Adoptively transferred murine islet-specific EngT_regs homed to the pancreas and blocked diabetes triggered by islet-specific T_effs or diabetogenic polyclonal T_effs in recipient mice. These data demonstrate the potential of antigen-specific EngT_regs as a targeted therapy for preventing T1D.