非小细胞肺癌患者外周血及肿瘤组织中CD4^+T淋巴细胞上Tim-3的表达及其对巨噬细胞活性的影响

目的研究非小细胞肺癌(NSCLC)患者外周血及肿瘤组织中CD4^+T淋巴细胞上T细胞免疫球蛋白黏蛋白结构域分子3(Tim-3)的表达及其临床意义。方法收集西南医科大学附属医院2017年1月至2018年6月收治住院的64例NSCLC患者外周血、肺癌组织和癌旁正常肺组织标本,同时选取40例健康人群作为对照。采用流式法检测Tim-3和程序性死亡蛋白配体(PD-1)在CD4^+T细胞上的表达,实时荧光定量PCR(qPCR)测定巨噬细胞诱导型一氧化氮合酶(iNOS)和精氨酸酶1(Arg1)mRNA的水平,分析Tim-3和PD-1与巨噬细胞活性的关系。结果肺癌患者外周血中CD4^+T淋巴细胞上Tim-3、PD-1表达分别为4.96%和24.13%,均显著高于健康对照人群的2.05%及10.25%(P<0.05);且肺癌患者外周血CD4^+T淋巴细胞上Tim-3^+PD-1^+的表达水平为4.36%,也明显高于健康对照组的2.63%(P<0.05)。肺癌组织中CD4^+T淋巴细胞上Tim-3、PD-1表达及Tim-3^+PD-1^+共表达均显著高于癌旁正常组织,差异均有统计学意义(P<0.05)。与癌旁正常组织比较,肺癌组织中iNOS mRNA略有降低,而Arg1 mRNA水平则显著升高,且肺组织中Tim-3、PD-1表达和Tim-3^+PD-1^+共表达与巨噬细胞活性iNOS/Arg1比值均呈负相关(P<0.05)。结论NSCLC患者外血周及肿瘤组织中CD4^+T淋巴细胞上Tim-3和PD-1表达上调,促使M1型巨噬细胞转变为M2型,具有负性免疫调节作用。

Tim-3 expression of CD4^+T lymphocytes in peripheral blood and tumor tissue of patients with NSCLC and its effect on macrophage activity

Objective To investigate the T-cell immunoglobulin and mucin-domain-containing molecule-3(Tim-3)expression of CD4^+T lymphocytes in peripheral and tumor tissue of patients with non-small cell lung cancer(NSCLC)and its clinical significance.Methods Samples of peripheral blood,lung cancer tissue and adjacent normal lung tissue of 64 NSCLC patients admitted and hospitalized in Affiliated Hospital of Southwest Medical University were collected from January 2017 to June 2018.Meanwhile,40 cases of healthy people were chosen as control.Flow cytometry was used to detect the expression of Tim-3 and PD-1 on CD4^+T cells,and the content of iNOS and Arg1 mRNA in macrophages was determined by qPCR to analyze the relationship between Tim-3 and PD-1 and the activity of macrophages.Results The percentages of Tim-3 and PD-1 expression on CD4^+T lymphocytes in peripheral blood of lung cancer patients were 4.96%and 24.13%,which were significantly higher than those in healthy control groups(2.05%and 10.25%,P<0.05).The Tim-3^+PD-1^+expression level of CD4^+T lymphocytes in peripheral blood of lung cancer patients was 4.36%,which was also significantly higher than 2.63%in the healthy control group(P<0.05).The percentages of Tim-3,PD-1 and Tim-3^+PD-1^+expression on CD4^+T lymphocytes in lung cancer tissue were significantly higher than that of normal tissue adjacent to carcinoma with statistically significant differences(P<0.05).Compared with normal tissue adjacent to carcinoma,iNOS mRNA in lung cancer tissues was slightly lower,but Arg1 mRNA levels were significantly increased and Tim-3,PD-1 expression and Tim-3^+PD-1^+expression in the lung tissue were negatively correlated with the activity of macrophages iNOS/Arg1 ratio(P<0.05).Conclusion Tim-3 and PD-1 expression of CD4^+T cells in the peripheral blood and tumor tissues of NSCLC patients is upregulated,and prompted the M1 to M2 macrophages,which presents negative immune regulating effect.