宫颈鳞癌患者外周血和肿瘤组织中PD-1/PD-L1及相关免疫细胞的变化特点及其临床意义

Characteristics and clinical significance of the changes in PD-1/PD-L1 and related immune cells in peripheral blood and tumor tissues of patients with cervical squamous cell cancer

目的:探讨PD-1/PD-L1通路及相关免疫细胞在宫颈鳞癌(cervical squamous cell cancer,CSCC)发生、发展中的变化特点及其临床意义。方法:收集2018年12月至2020年9月在福州市第一医院接受手术的CSCC患者和健康体检人员的癌组织/宫颈组织和外周血样本,分为健康对照组、宫颈上皮内癌变(cervical intraepithelial neoplasia,CIN)Ⅱ级组、CINⅢ级组和CSCC组,代表CSCC发生、发展进程各阶段,每组50例。ELISA法检测各组人员的外周血血浆中PD-1、PD-L1、叉头状转录因子P3(FOXP3)的表达水平,FCM法检测各组人员外周血PD-1^(+)CD4^(+)CD25^(+)CD127^(-/low)细胞的数量,应用多色荧光组织染色法检测肿瘤浸润性淋巴细胞(TIL)在CSCC组织中的分布特点。结果:随着模拟的CSCC发生和发展,外周血中PD-1、PD-L1和FOXP3的表达呈上升趋势,术后则呈下降趋势。在CSCC患者抗凝全血中,CD4^(+)、CD4^(+)CD25^(+)CD127^(-/low)以及PD-1^(+)CD4^(+)CD25^(+)CD127^(-/low)细胞占淋巴细胞的比例增加。在CSCC组织中可见大量CD4^(+)、CD8^(+)和FOXP3^(+)细胞浸润,其中CD4^(+)和FOXP3^(+)细胞主要围绕肿瘤细胞聚集区分布、CD8^(+)和PD-L1^(+)细胞则呈广泛弥漫性分布。结论:PD-1、PD-L1、FOXP3和适应性调节性T细胞是促进CSCC发生发展的重要因素,其可作为ESCC免疫治疗的靶点和临床预后的潜在标志物。

Objective:To investigate the changes in PD-1/PD-L1 pathway and related immune cells in the occurrence and development of cervical squamous cell cancer(CSCC)and their clinical significance.Methods:Cervical tissue/cancerous tissue and peripheral blood samples were collected from CSCC patients who underwent surgery and healthy controls who had physical examination at the First Hospital of Fuzhou City from December 2018 to September 2020 and were divided into healthy control group,cervical intraepithelial neoplasia(CIN)gradeⅡgroup,CIN gradeⅢgroup,and CSCC group to represent the occurrence and development of CSCC(n=50 in each group).The expression levels of PD-1,PD-L1 and forkhead box transcription factor P3(FOXP3)in the peripheral blood of each group were detected by ELISA,the numbers of PD-1;CD4;CD25;CD127;cells in the peripheral blood of each group were detected by FCM,and the distribution of tumor-invasive lymphocytes(TIL)in CSCC tissues was detected by multicolor fluorescent immunohistochemistry.Results:The expression of PD-1,PD-L1 and FOXP3 in peripheral blood tended to increase with the onset and progression of mock CSCC and to decrease after surgery.The proportion of CD4;,CD4;CD25;CD127;and PD-1;CD4;CD25;CD127;cells in lymphocytes increased in the anticoagulated whole blood of CSCC patients.Massive infiltration of CD4;,CD8;and FOXP3;cells was observed in CSCC tissues,with CD4;and FOXP3;cells mainly distributed around tumor cells and CD8;and PD-L1;cells widely and diffusely distributed.Conclusion:PD-1,PD-L1,FOXP3 and adaptive regulatory T cells are important factors contributing to the development of CSCC,which can be used as potential immunotherapeutic targets and potential prognostic markers for CSCC.