肝细胞癌TACE术后肿瘤内T细胞表型与CD73的表达及其与预后的相关性

Changes of intratumoral T cell phenotype and CD73 expression in hepatocellular carcinoma after trans-arterial chemoembolization and its value in predicting prognosis

目的探讨TACE术后肿瘤微环境的T细胞表型、PD-1(程序性细胞死亡受体)及其配体PD-L1和免疫抑制相关蛋白CD73表达变化及其在预测预后中的价值。方法本研究收集了20例2019年5月到2020年12月间经TACE治疗后转化切除的肝癌组织样本(TACE组,简称T+组),同时收集20例同期未接受过TACE治疗的肝癌组织样本作为对照组(无TACE组,简称T-组)。通过对肿瘤组织切片进行多重荧光免疫组化,比较两组患者肿瘤内(FOXP3^(+))调节性T细胞(Treg)、免疫耗竭的(CD8^(+)/PD-1^(+))T细胞数量及PD-L1和CD73的表达情况。统计分析不同的T细胞表型及CD73表达水平对患者无进展生存期的影响,并通过Cox多因素回归分析影响患者预后的主要因素。结果T+组中的肿瘤组织中的CD73表达水平明显高于T-组(11.86 vs 7.7,P=0.047)。T+组的Treg、CD8^(+)和CD8^(+)PD-1^(+)T细胞数量分别为35.93、86.09、13.91,明显低于T-组的60.02(P=0.013)、120.27(P=0.043)和28.36(P=0.015),而PD-L1表达水平在T+与T-中无明显差异(23.68 vs 21.04,P=0.380)。CD73高表达组显示更少的CD8^(+)(84.83 vs 121.53,P=0.029)和CD8^(+)PD-1^(+)(15.00 vs 27.27,P=0.041)T细胞数量,FOXP3^(+)数量两组无差异(45.50 vs 50.45,P=0.623)。Kaplan-Meier生存分析显示,CD73高表达以及CD8^(+)PD-1^(+)高表达患者的无进展生存期更差(P值分别为0.009和0.019)。结论TACE术后肿瘤免疫微环境CD73表达升高,而FOXP3^(+)和CD8^(+)/PD-1^(+)T细胞数量降低,而CD73高表达以及CD8^(+)PD-1^(+)高表达与患者不良预后密切相关,提示靶向CD73可能会成为TACE联合治疗的新方向。

Objective To study the changes of T cell phenotype,PD-1(programmed cell death receptor),PD-L1 and immunosuppression-related protein CD73 expression in tumor microenvironment after trans-arterial chemoembolization(TACE)and its value in predicting prognosis.Methods Hepatocellular carcinoma tissue samples from 20 patients with TACE(T+group)and 20 without TACE(T-group)prior to surgery were collected from May 2019 to December 2020.We profiled the number of regulatory(FOXP3^(+))Treg,immune-exhausted(CD8^(+)/PD-1^(+))T cells and PD-L1,CD73 expression in the tumor by multiple fluorescence immunohistochemistry on tumor tissue sections.We profiled the effects of different T cell phenotypes and CD73 expression levels on progression-free survival.The main factors affecting the prognosis of patients were analyzed by multivariate Cox regression.Results The expression level of CD73 in tumor tissues in the T+group was higher than that in T-group(11.86 vs 7.7,P=0.047).The numbers of Treg,CD8^(+)and CD8^(+)PD-1^(+)T cells in the T+group were 35.93,86.09,and 13.91,which were lower than that in T-group of 60.02(P=0.013),120.27(P=0.043),and 28.36(P=0.015),while the expression level of PD-L1 was not different between T+group and T-group(23.68 vs 21.04,P=0.380).The CD73 high expression group showed lower CD8^(+)(84.83 vs 121.53,P=0.029)and CD8^(+)PD-1^(+)(15.00 vs 27.27,P=0.041)T cell numbers,and there was no difference in the number of FOXP3^(+)between the two groups(45.50 vs 50.45,P=0.623).Kaplan-Meier survival analysis showed that patients with high CD73 expression and high CD8^(+)PD-1^(+)expression had worse progression free survival(P=0.009 and 0.019,respectively).Conclusions After TACE,the expression of CD73 in the tumor immune microenvironment increased,while the number of FOXP3^(+)and CD8^(+)/PD-1^(+)T cells decreased.The high expression of CD73 and the low expression of CD8^(+)PD-1^(+)were related to the poor prognosis of patients,suggesting that targeting CD73 may become a new idea of TACE combination therapy.