青蒿琥酯上调肺癌患者CD4^(+)和CD8^(+)T淋巴细胞亚群CD39、CD279和颗粒酶B表达

Artesunate upregulates the expression of CD39,CD279 and granzyme B in CD4^(+)and CD8^(+)T lymphocytes of patients with lung cancer

目的探讨青蒿琥酯(ART)对肺癌患者T淋巴细胞免疫功能的影响。方法随机选取健康体检者15例(NC组),肺癌患者21例(LC组),收集临床信息,采集并分离外周血单个核细胞(PBMC)。用ART处理PBMC 24h后,流式细胞术(FCM)检测ART的半数杀伤浓度(LC_(50))及诱导T细胞高表达CD39和CD279的最佳浓度;最佳浓度的ART诱导后,FCM检测NC组CD8^(+)和CD4^(+)T细胞CD39、CD279的表达水平,以及LC组CD8^(+)和CD4^(+)T细胞CD39、CD279、CD38、CD28、颗粒酶B(GrzB)、穿孔素(PerF)、γ干扰素(IFN-γ)和白细胞介素2(IL-2)的水平。结果ART的LC_(50)为522μmol/L,最佳浓度为200μmol/L。LC组CD8^(+)和CD4^(+)T细胞CD279的基线表达显著高于NC组;200μmol/L ART诱导后,在NC组CD8^(+)和CD4^(+)T细胞中,CD39的表达显著升高;在LC组CD8^(+)和CD4^(+)T细胞中,CD39、CD279和GrzB的表达显著升高,IL-2表达显著降低,而CD38、CD28、IFN-γ和PerF等表达未见显著差异。促进ART诱导CD8^(+)T细胞表达CD39的临床因素有未放疗;促进ART诱导CD8^(+)T细胞表达CD279的因素有年龄>60岁、淋巴细胞数>1.26×10^(9)/L、中性粒细胞/淋巴细胞比率(NLR)<5、放疗、0.29×10^(9)/L≤单核细胞数≤0.95×10^(9)/L。结论肺癌患者T淋巴细胞高表达CD279;ART诱导肺癌患者CD8^(+)和CD4^(+)T细胞CD39、CD279和GrzB的表达上调,从而调节T细胞亚群的免疫功能。

Objective To investigate the effect of artesunate(ART)on T lymphocyte immune function in patients with lung cancer.Methods Fifteen healthy people(NC group)and twenty-one lung cancer patients(LC group)were randomly selected to collect their clinical information and isolate peripheral blood mononuclear cells(PBMCs).After 24 hour-treatment of PBMCs with ART,the median lethal concentration(LC_(50))and the optimal concentration of ART induced high expression of CD39 and CD279 in T cell membrane were determined by flow cytometry(FCM).Following the induction of ART with the best concentration,the expression levels of CD39 and CD279 on CD8^(+)and CD4^(+)T cells in NC group,and the expression levels of CD39,CD279,CD38,CD28,granzyme B(GrzB),perforin(PerF),interferonγ(IFN-γ)and interleukin-2(IL-2)on CD8^(+)and CD4^(+)T cells in LC group were detected by FCM.Results LC_(50) and optimal concentration of ART were 522μmol/L and 200μmol/L,respectively.Compared with the NC group,the baseline expression levels of CD279 on CD8^(+)and CD4^(+)T cells in LC group was significantly higher.Moreover,the expression levels of CD39 increased significantly after inducing 200μmol/L ART,in the CD8^(+)and CD4^(+)T cell of NC groups;In CD8^(+)and CD4^(+)T cells of LC group,the expression of CD39,CD279 and GrzB increased significantly,while that of IL-2 decreased markedly.No significant difference was detected in the expression levels of CD38,CD28,IFN-γand PerF.The clinical factors that promote the expression of CD39 on CD8^(+)T cells induced by ART showed no radiotherapy.The clinical factors that promote the expression of CD279 on CD8^(+)T cells induced by ART include age>60 years old,lymphocyte count>1.26×10~9/L,NLR<5,radiotherapy,0.29×10~9/L≤monocyte count≤0.95×10~9/L.Conclusion The expression of CD279 on T lymphocytes is higher in lung cancer patients;ART induces the upregulation of CD8^(+)and CD4^(+)T cells CD39,CD279 and GrzB in lung cancer patients,thus regulating the immune function of T cell subsets.