EBV阳性鼻咽癌相关性中性粒细胞抑制肿瘤微环境中的CD8^(+)T细胞活化  

Tumor-associated neutrophils in EBV-positive nasopharyngeal carcinoma suppress the activation of CD8^(+)T cells in tumor microenvironment

作　　者：欧阳蒂君 陈楠[1] 杨洁莹 陈媛媛[2] 向橦 夏建川[2] OUYANG Dijun;CHEN Nan;YANG Jieying;CHEN Yuanyuan;XIANG Tong;XIA Jianchuan(Department of Oncology,The Fifth Affiliated Hospital of Sun Yat-sen University,Zhuhai 519000,Guangdong,China;Department of Experimental Research,State Key Laboratory of Oncology in South China,Collaborative Innovation Center for Cancer Medicine,Guangdong Key Laboratory of Nasopharyngeal Carcinoma Diagnosis and Therapy,Sun Yat-sen University Cancer Center,Guangzhou 510060,Guangdong,China)

机构地区：[1]中山大学附属第五医院肿瘤科,广东珠海519000 [2]中山大学肿瘤防治中心实验研究部,华南肿瘤学国家重点实验室,肿瘤医学省部共建协同创新中心,广东省鼻咽癌诊断与治疗重点实验室,广东广州510060

出　　处：《中国肿瘤生物治疗杂志》2022年第5期399-409,共11页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81874129)。

摘　　要：目的:分析鼻咽癌(NPC)肿瘤微环境(TME)中的肿瘤相关中性粒细胞(TAN)浸润与患者的预后和临床病理特征之间的相关性,初步探讨EBV阳性NPC的TAN对CD8^(+)T细胞活化的作用。方法:收集2008年至2012年间中山大学肿瘤防治中心收治的118例初治且无转移的EBV阳性NPC患者的肿瘤组织标本,通过免疫组织化学染色法观察并分析NPC组织中CD8^(+)T细胞、TAN浸润和EBV感染之间的关系,检测NPC组织标本中的TAN浸润程度并分析其与患者预后和临床病理特征之间的关系;流式细胞术检测HK1-EBV细胞培养上清液对TAN极化的N2型标志物(CD182和CD206)表达水平的影响,并进一步检测极化后的TAN对CD8^(+)T细胞活化标志物(CD69和PD-1)表达水平的影响。结果:EBV阳性NPC组织中CD8^(+)T细胞和TAN浸润增多且两者在数量上呈负相关关系(P=0.005 2);EBV阳性NPC组织中高水平浸润的TAN与患者的不良预后密切相关(OS:P=0.025,PFS:P=0.027),TAN浸润水平是EBV阳性NPC患者总生存时间(P=0.035)的独立预后因子;与对照组相比,HK1-EBV细胞培养上清液可诱导TAN极化并高表达N2型标志物(CD182:P<0.001;CD206:P<0.01);极化后的TAN可抑制CD8^(+)T细胞CD69的表达并促进PD-1的表达(P<0.001)。结论:EBV阳性NPC能够促进TME中的TAN浸润增多并使其极化成N2型,从而抑制CD8+T细胞的活化,发挥免疫抑制作用,与NPC患者的不良预后密切相关。Objective: To analyze tumor-associated neutrophils(TANs) infiltration in EBV-positive nasopharyngeal carcinoma(NPC) tumor microenvironment(TME) and its correlation with the prognosis and clinicopathological features of patients with NPC, and to preliminarily investigate the role of TANs in activating CD8^(+)T cells in EBV-positive NPC. Methods: Tumor tissue specimens from 118patients with EBV-positive NPC without metastases admitted to Sun Yat-sen University Cancer Center from 2008 to 2012 were collected. Immunohistochemistry was performed to analyze CD8^(+)T cells infiltration and TANs infiltration in NPC and their correlation with EBV infection, and to evaluate the infiltration of TANs in 118 human EBV-positive NPC tissue samples and analyze its correlation with the prognosis and clinicopathological features of NPC patients;flow cytometry was used to detect the expression of N2 marker(CD182 and CD206) of neutrophils stimulated by HK1-EBV-conditioned medium and the expression of CD69 and PD-1 of CD8+T cells co-cultured with TANs. Results: There were more CD8^(+)T cells and neutrophils infiltrating in EBV-positive NPC where they were negatively correlated(P=0.0052);densely infiltrated TANs in EBV-positive NPC tissues were positively correlated with poor prognosis of patients(P=0.025 in OS;P=0.027 in PFS);the infiltration of TANs was proved to be an independent risk factor for overall survival(P=0.035) in patients with NPC;compared with Control group, freshly isolated neutrophils cultured in HK1-EBV-conditioned medium showed an increased expression of N2 markers(P<0.001 in CD182;P<0.01 in CD206) which could suppress the expression of CD69 and promote the expression of PD-1 of CD8^(+)T cells. Conclusion: More TANs infiltrate in EBV-positive NPC where they polarize into N2 neutrophils, exerting an immunosuppressive effect by suppressing the activation of CD8^(+)T cells and having a positive correlation with the poor prognosis of NPC patients.

关 键 词：EB病毒 鼻咽癌 肿瘤相关中性粒细胞 CD8^(+)T细胞 肿瘤微环境 

分 类 号：R739.6[医药卫生—肿瘤] R730.3[医药卫生—临床医学]