Adenosine Generated by Regulatory T Cells Induces CD8(+) T Cell Exhaustion in Gastric Cancer through A2aR Pathway

BACKGROUND: Adenosine, derived from the degradation of ATP via ectonucleotidases CD39 and CD73, is a critical immunosuppressive metabolite in the hypoxic microenvironment of tumor tissue. Adenosine signaling via A2aR can inhibit the antitumor immune response of CD8(+) T cells. CD39 and CD73 high-expressing Tregs play a critical role in tumor immune evasion of gastric cancer (GC). The present study investigated the underlying mechanism by which Tregs suppress antitumor immune responses in GC. MATERIALS AND METHODS: Fifty-two GC samples were collected, and the frequency of FoxP3(+) Tregs and CD8(+) T cells and density ratios of A2aR(+)/CD8(+) T cells, CD39(+)/FoxP3(+) Tregs, and CD73(+)/FoxP3(+) Tregs in GC were assessed with multiplex immunofluorescence. The expression of FoxP3 and A2aR in GC tissues was also detected by the immunoblotting assay. We next investigated the relationship between density of FoxP3(+) Tregs, ratio of A2aR(+)/CD8(+) T cells, and clinicopathological parameters. At the same time, Tregs and CD8(+) T cells were isolated from peripheral blood of five GC patients, and the antagonists of CD39 and CD73 were used to assess the ability of Tregs to decompose ATP into adenosine. In addition, we cocultured CD8(+) T cells and Tregs with antagonists of A(2a)R and A(2b)R in order to examine the alterations in immune function of CD8(+) T cells. RESULTS: The density of both FoxP3(+) Tregs and A2aR(+)/CD8(+) T cells was higher in GC tissue compared to peritumoral normal tissue and significantly correlated with the TNM stage, lymph node metastasis, and distant metastasis of GC. The process of Treg hydrolysis of ATP into adenosine was blocked by the antagonists of CD39 and CD73. In addition, Tregs could induce apoptosis and inhibit proliferation of CD8(+) T cells, while this effect could be obviously reduced by applying the antagonist of A(2a)R or A(2a)R(+)A(2b)R. Moreover, IFN-γ, TNF-α, and perforin generated by CD8(+) T cells could also be inhibited through the adenosine A2aR pathway. CONCLUSIONS: The FoxP3(+) Tregs and A2aR(+)/CD8(+) T cells were excessively infiltrated in GC tissue. Tregs from GC can decompose ATP to adenosine and in turn induce apoptosis and inhibit the proliferation of CD8(+) T cells through the A2aR pathway, further leading to immune escape of GC.