Novel Effector Phenotype of Tim-3+ Regulatory T Cells Leads to Enhanced Suppressive Function in Head and Neck Cancer Patients

Purpose: Regulatory T (Treg) cells are important suppressive cells among tumor-infiltrating lymphocytes (TIL). Treg cells express the well-known immune checkpoint receptor PD-1, which is reported to mark exhausted Treg with lower suppressive function. T-cell immunoglobulin mucin (Tim)-3, a negative regulator of Th1 immunity, is expressed by a sizeable fraction of TIL Tregs, but the functional status of Tim-3(+) Tregs remains unclear. Experimental Design: CD4(+)CTLA-4(+)CD25(high) Treg cells were sorted from freshly excised head and neck squamous cell carcinoma (HNSCC) TIL based on Tim-3 expression. Functional and phenotypic features of these Tim-3(+) and Tim-3 similar to TIL Tregs were tested by in vitro suppression assays and multi-color flow cytometry. Gene-expression profiling and NanoString analysis of Tim-3(+) TIL Treg were performed. A murine HNSCC tumor model was used to test the effect of anti-PD-1 immunotherapy on Tim-3(+) Treg. Results: Despite high PD-1 expression, Tim-3(+) TIL Treg displayed a greater capacity to inhibit naive T-cell proliferation than Tim-3(+) Treg. Tim-3(+) Treg from human HNSCC TIL also displayed an effector-like phenotype, with more robust expression of CTLA-4, PD-1, CD39, and IFN-gamma receptor. Exogenous IFN-gamma treatment could partially reverse the suppressive function of Tim-3(+) TIL Treg. AntiPD- 1 immunotherapy downregulated Tim-3 expression on Tregs isolated from murine HNSCC tumors, and this treatment reversed the suppressive function of HNSCC TIL Tregs. Conclusions: Tim-3(+) Treg are functionally and phenotypically distinct in HNSCC TIL, and are highly effective at inhibiting T-cell proliferation despite high PD-1 expression. IFN-gamma induced by anti-PD-1immunotherapymay be beneficial by reversing Tim-3(+) Treg suppression. (C) 2018 AACR.