CD4⁺ T cell activation and concomitant mTOR metabolic inhibition can ablate microbiota-specific memory cells and...

Microbiota-reactive CD4⁺ T memory (T_M) cells are generated during intestinal infections and inflammation, and can revert to pathogenic CD4⁺ T effector (T_E) cells, resulting in chronicity of inflammatory bowel disease (IBD). Unlike T_E cells, T_M cells have a low rate of metabolism unless they are activated by reencountering cognate antigen. Here, we show that the combination of cell activation and metabolic checkpoint inhibition (CAMCI), by targeting key metabolic regulators mTORC and AMPK, resulted in cell death and anergy, but enhanced the induction of the regulatory subset. Parenteral application of this treatment with a synthetic peptide containing multiple flagellin T cell epitopes (MEP1) and metabolic inhibition successfully prevented the development of CD4⁺ T cell-driven colitis. Microbiota-specific CD4⁺ T cells, especially the pathogenic T_E subsets, were decreased 10-fold in the intestinal lamina propria. Furthermore, using the CAMCI strategy, we were able to prevent antigen-specific T_M cell formation upon initial antigen encounter, and ablate existing T_M cells upon reactivation in mice, leading to an altered transcriptome in the remaining CD4⁺ T cells after ablation. Microbiota flagellin-specific CD4⁺ T cells from patients with Crohn's disease were ablated in a similar manner after CAMCI in vitro, with half of the antigen-specific T cells undergoing cell death. These results indicate that parenteral activation of microbiota-specific CD4⁺ T cells with concomitant metabolic inhibition is an effective way to ablate pathogenic CD4⁺ T_M cells and to induce T regulatory (T_reg) cells that provide antigen-specific and bystander suppression, supporting a potential immunotherapy to prevent or ameliorate IBD.