Li Lifeng, Yang Li, Wang Liping, Wang Fei, Zhang Zhen, Li Jieyao, Yue Dongli, Chen Xinfeng, Ping Yu, Huang Lan, Zhang Bin, Zhang Yi
Biotherapy Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, People's Republic of China.
Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, People's Republic of China.
Int J Cancer. 2016 Nov 15;139(10):2261-9. doi: 10.1002/ijc.30289. Epub 2016 Aug 6.
Malignant pleural effusion (MPE) is an indication of advanced cancer. Immune dysfunction often occurs in MPE. We aimed to identify the reason for impaired T cell activity in MPE from lung cancer patients and to provide clues toward potential immune therapies for MPE. The surface inhibitory molecules and cytotoxic activity of T cells in MPE and peripheral blood (PB) were analyzed using flow cytometry. Levels of inflammatory cytokines in MPE and PB were tested using ELISA. TGF-β expression in tumor-associated macrophages (TAMs) was also analyzed. The effect of TAMs on T cells was verified in vitro. Lastly, changes in T cells were evaluated following treatment with anti-TGF-β antibody. We found that expression levels of Tim-3, PD-1 and CTLA-4 in T cells from MPE were upregulated compared with those from PB, but levels of IFN-γ and Granzyme B were downregulated (p < 0.05). The amount of TGF-β was significantly higher in MPE than in PB (p < 0.05). TGF-β was mainly produced by TAMs in MPE. When T cells were co-cultured with TAMs, expression levels of Tim-3, PD-1 and CTLA-4 were significantly higher than controls, whereas levels of IFN-γ and Granzyme B were significantly decreased, in a dose-dependent manner (p < 0.05). In vitro treatment with anti-TGF-β antibody restored the impaired T cell cytotoxic activity in MPE. Our results indicate that macrophage-derived TGF-β plays an important role in impaired T cell cytotoxicity. It will therefore be valuable to develop therapeutic strategies against TGF-β pathway for MPE therapy of lung cancer.
恶性胸腔积液（MPE）是晚期癌症的一种表现。免疫功能障碍在MPE中经常出现。我们旨在确定肺癌患者MPE中T细胞活性受损的原因，并为MPE的潜在免疫治疗提供线索。使用流式细胞术分析MPE和外周血（PB）中T细胞的表面抑制分子和细胞毒性活性。使用酶联免疫吸附测定（ELISA）检测MPE和PB中炎性细胞因子的水平。还分析了肿瘤相关巨噬细胞（TAM）中转化生长因子-β（TGF-β）的表达。在体外验证了TAM对T细胞的作用。最后，在用抗TGF-β抗体治疗后评估T细胞的变化。我们发现，与PB中的T细胞相比，MPE中T细胞上T细胞免疫球蛋白和粘蛋白结构域分子3（Tim-3）、程序性死亡受体1（PD-1）和细胞毒性T淋巴细胞相关蛋白4（CTLA-4）的表达水平上调，但γ干扰素（IFN-γ）和颗粒酶B的水平下调（p<0.05）。MPE中TGF-β的量显著高于PB（p<0.05）。TGF-β主要由MPE中的TAM产生。当T细胞与TAM共培养时，Tim-3、PD-1和CTLA-4的表达水平显著高于对照组，而IFN-γ和颗粒酶B的水平显著降低，呈剂量依赖性（p<0.05）。用抗TGF-β抗体进行体外治疗可恢复MPE中受损的T细胞细胞毒性活性。我们的结果表明，巨噬细胞衍生的TGF-β在T细胞细胞毒性受损中起重要作用。因此，开发针对TGF-β途径的治疗策略用于肺癌MPE治疗将具有重要价值。
