Targeting of Toll-like receptors inhibits CD4+ regulatory T cell function and activates lymphocytes in human PBMCs

摘要

Accumulating evidence suggests elements within tumors induce exhaustion of effector T cells and infiltration of immune-suppressive regulatory T cells (Tregs) thus preventing the development of durable anti-tumor immunity. Therefore, the discovery of agents that simultaneously block Treg suppressive function and reinvigorate effector function of lymphocytes is key to the development of effective cancer immunotherapy. Previous studies have shown that Toll-like receptor ligands (TLRL) could modulate the function of these T-cell targets; however, those studies relied on cell-free or accessory cell-based assay systems that do not accurately reflect in vivo responses. In contrast, we employed a human PBMC-based proliferation assay system to simultaneously monitor the effect of TLRLs on T cells (CD4⁺, CD8⁺, Tregs), B cells and NK cells, which gave different and even conflicting results. We found that the TLR7/8L:CL097 could simultaneously activate CD8⁺ T cells, B cells and NK cells plus block Treg suppression of T cells and B cells. The TLRLs TLR1/2L:Pam3CSK4, TLR5L:flagellin, TLR4L:LPS and TLR8/7L:CL075 also blocked Treg suppression of CD4⁺ or CD8⁺ T cell proliferation but not B cell proliferation. Besides CL097, TLR2L:PGN, CL075 and TLR9L:CpG-(A-C) were strong activators of NK cells. Importantly, we found that Pam3CSK4 could: 1) activate CD4+ T cells proliferation; 2) inhibit the expansion of IL-10⁺ nTregs and induction of IL-10⁺ CD4⁺ Tregs (Tr1); and 3) block nTreg suppressive function. Our results suggest these agents could serve as adjuvants to enhance the efficacy of current immunotherapeutic strategies in cancer patients.