Persistent exposure to tumor antigens results in exhausted tumor-infiltrating T cells (TILs) that express the immune checkpoint molecules, PD-1 and Tim3, and lack anti-tumor immunity. To examine the exhausted status of TILs in ovarian cancer, the potential for cytokine production, proliferation and cytotoxicity by purified PD-1⁺ Tim3⁺ CD8 TILs was assessed. The production of IFN-γ and TNF-α by PD-1⁺ Tim3⁺ CD8 TILs remained the same in an intracellular cytokine staining assay and was higher in a cytokine catch assay than that by PD-1⁻ Tim3⁻ and PD-1⁺ Tim3⁻ CD8 TILs. %Ki67⁺ was higher in PD-1⁺ Tim3⁺ CD8 TILs than in PD-1⁻ Tim3⁻ CD8 TILs. However, patients with high PD-1⁺ Tim3⁺ CD8 TILs had a poor prognosis. The potential for cytotoxicity was then examined. %Perforin⁺ and %granzyme B⁺ were lower in PD-1⁺ Tim3⁺ CD8 TILs than in PD-1⁻ Tim3⁻ and PD-1⁺ Tim3⁻ CD8 TILs. To observe the potential for direct cytotoxicity by T cells, a target cell line expressing membrane-bound anti-CD3scFv was newly established and a cytotoxic assay targeting these cells was performed. The cytotoxicity of PD-1⁺ Tim3⁺ CD8 TILs was significantly lower than that of PD-1⁻ Tim3⁻ and PD-1⁺ Tim3⁻ CD8 TILs. Even though PD-1⁺ Tim3⁺ CD8 TILs in ovarian cancer showed a sustained potential for cytokine production and proliferation, cytotoxicity was markedly impaired, which may contribute to the poor prognosis of patients with ovarian cancer. Among the impaired functions of exhausted TILs, cytotoxicity may be an essential target for cancer immunotherapy.

中文翻译：

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PD-1 + Tim3 +肿瘤浸润性CD8 T细胞维持产生IFN-γ的潜力，但在卵巢癌中丧失细胞毒活性。

长期暴露于肿瘤抗原会导致耗尽的肿瘤浸润性T细胞（TIL）表达免疫检查点分子PD-1和Tim3，并且缺乏抗肿瘤免疫力。为了检查卵巢癌中TIL的衰竭状态，评估了纯化的PD-1 ⁺ Tim3 ⁺ CD8 TIL对细胞因子产生，增殖和细胞毒性的潜力。通过PD-1的产生IFN-γ和TNF-α的⁺ TIM3 ⁺ CD8的TIL仍留在细胞内细胞因子染色测定法相同，并且在细胞因子捕获测定比由PD-1是高^- TIM3 ^-和PD-1 ⁺ Tim3 ^- CD8 TIL。PD-1 ^+中％Ki67 ⁺较高Tim3 ⁺ CD8 TIL比PD-1 ^- Tim3 ^- CD8 TIL中的高。然而，PD-1 ⁺ Tim3 ⁺ CD8 TILs高的患者预后较差。然后检查细胞毒性的潜力。％穿孔⁺和％粒酶B ⁺在PD-1均较低⁺ TIM3 ⁺ CD8的TIL比PD-1 ^- TIM3 ^-和PD-1 ⁺ TIM3 ^- CD8的TIL。为了观察T细胞直接细胞毒性的可能性，新建立了表达膜结合抗CD3scFv的靶细胞系，并进行了针对这些细胞的细胞毒性测定。PD-1的细胞毒性⁺ TIM3 ⁺ CD8的TIL比PD-1的被显著降低^- TIM3 ^-和PD-1 ⁺ TIM3 ^- CD8的TIL。尽管PD-1 ⁺ Tim3 ⁺ CD8 TILs在卵巢癌中显示出持续的细胞因子产生和增殖潜能，但细胞毒性明显受损，这可能导致卵巢癌患者预后不良。在疲惫的TIL的功能受损中，细胞毒性可能是癌症免疫治疗的重要靶标。