Tim3/Gal9 interactions between T cells and monocytes result in an immunosuppressive feedback loop that inhibits Th1 responses in osteosarcoma patients

Abstract

The Tim3/Gal9 pathway is associated with immunosuppression and worse clinical outcome in multiple cancers. To illustrate the specific mechanism of Tim3/Gal9 interaction in osteosarcoma, we examined expression, function, and regulation of Tim3/Gal9 in various cells from osteosarcoma patients. Data showed that CD4⁺ T cells, CD8⁺ T cells, and monocytes from both peripheral blood and tumor of osteosarcoma patients contained high frequencies of Tim3⁺ cells, while the Gal9 expression was primarily found in regulatory T cells (Tregs) from osteosarcoma patients and was elevated compared to that in non-cancer controls. The Tim3⁺ CD4⁺ and CD8⁺ T cells presented lower proliferation capacity compared to their Tim3^- counterparts, which could be reverted by blocking Tim3 or Gal9. Interestingly, purified Tim3⁺ CD4⁺ T cells secreted more interferon gamma (IFNγ) than purified Tim3^- CD4⁺ T cells, but IFNγ production by Tim3⁺ CD4⁺ T cells was vulnerable to Gal9-mediated suppression. In monocytes, Tim3 expression was associated with high interleukin (IL)-10 and low IL-12 cytokine secretion profile. Exogenous recombinant Gal9, as well as CD4⁺CD25⁺ Treg supernatant, further decreased IL-12 expression in monocytes. In CD4⁺ T cell-monocyte coculture experiments, Tim3⁺ monocytes inhibited IFNγ expression from total CD4⁺ T cells and the development of IFNγ response in naive CD4⁺ T cells. Blocking the Tim3/Gal9 pathway reverted these effects. Together, these results suggested that in osteosarcoma patients, Tim3 expression did not directly mediate immune suppression, but the interaction between Tim3⁺ T cells and monocytes, naive CD4⁺ T cells, and Gal9-expressing CD4⁺CD25⁺ Tregs could resulting in progressive suppression of Th1 responses.

Keywords: Gal9; Osteosarcoma; Tim3.