CD45RO^(+)记忆T细胞作为非小细胞肺癌患者预后标志物的研究

CD45RO^(+) Memory T Lymphocytes As A Candidate Marker for Non-small Cell Lung Cancer

背景与目的肺癌是世界范围内最常见的恶性肿瘤之一。肿瘤微环境中多种多样的免疫浸润细胞,是肿瘤免疫的重要组成,对患者预后具有临床意义。CD45RO^(+)肿瘤浸润淋巴细胞(tumor infiltrating lymphocytes,TILs),即记忆T细胞,其表达与多种肿瘤预后相关。本研究旨在探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)中评估肿瘤和基质区CD45RO^(+)TILs密度与患者临床特征和预后的关系,及其联合程序性死亡受体配体1(programmed cell deathligand 1,PD-L1)作为预后标志物的临床价值。方法对167例NSCLC患者的组织微阵列进行多重荧光免疫组织化学染色,标记CD45RO、细胞角蛋白(cytokeratin,CK)和PD-L1。利用人工智能图像识别技术和肿瘤细胞特异性CK染色,划分组织中的肿瘤区和基质区,评估肿瘤区和基质区CD45RO^(+)TILs的密度以及肿瘤细胞的PD-L1表达水平。采用非参检验分析CD45RO^(+)TILs与患者临床特征的关系,使用Kaplan-Meier方法和Cox风险比例模型分析CD45RO^(+)TILs独立或与PD-L1联合与肿瘤预后的关系。结果CD45RO^(+)TILs的密度与患者年龄、吸烟、肿瘤分期和病理类型显著相关。在NSCLC和肺腺癌(lung adenocarcinoma,LUAD)患者中,基质区高密度CD45RO^(+)TILs具有更长的总生存期(overall survival,OS)(NSCLC:P=0.007;LUAD:P<0.001),并且是OS的独立预后因素(NSCLC:HR=0.559,95%CI:0.377-0.829,P=0.004;LUAD:HR=0.352,95%CI:0.193-0.641,P=0.001)。联合肿瘤细胞的PD-L1评分以及所有区域CD45RO^(+)TILs的浸润评分将患者分为四组:其中PD-L1^(+)/CD45RO^(+)患者无病生存期(disease-free survival,DFS)最长,PD-L1^(+)/CD45RO-的患者DFS时间最短,并可作为DFS预后的独立因素(HR=2.221,95%CI:1.258-3.919,P=0.006)。结论肿瘤组织中CD45RO^(+)TILs密度以及CD45RO^(+)TILs联合肿瘤区PD-L1,与NSCLC的临床病理特征及预后显著相关,可作为新的生存预后标志物。

Background and objective Lung cancer is the most common malignancy world-wide.There are a variety of immune infiltrating cells in tumor microenvironment,which is an important component of tumor immunity and has clinical significance for the prognosis of patients.CD45RO is a surface marker of memory T cells.The expression of CD45RO+tumor infiltrating lymphocytes(TILs)is associated with the prognosis of many tumors.The purpose of this study was to evaluate the relationship between the density of CD45RO+TILs in tumor and stromal area and the clinical characteristics of patients with non-small cell lung cancer(NSCLC)and its impact on the prognosis of patients.We aimed to explore the clinical value of CD45RO+TILs and programmed cell death ligand 1(PD-L1)as prognostic markers.Methods Multiple fluorescent immunohistochemical staining was used to stain the tissue microarray chips of 167 patients with NSCLC,marking CD45RO,cytokeratin(CK)and PD-L1.Using artificial intelligence image recognition technology and tumor cell-specific CK staining,divide the tumor and stromal area in the tissue,evaluate the density of CD45RO+TILs in the tumor and stromal area,and the expression level of PD-L1 in tumor cells.The non-parametric test was used to analyze the relationship between CD45RO+TILs and the clinical characteristics of patients,and the Kaplan-Meier method and Cox risk ratio model were used to analyze the relationship between CD45RO+TILs independently or in combination with PD-L1 and tumor prognosis.Results The density of CD45RO+TILs was significantly associated with patient age,smoking,tumor stage,and pathological type.Single-factor survival analysis showed that NSCLC(P=0.007)stromal region and lung adenocarcinoma(LUAD)(P<0.001)with CD45RO+TILs high density had better OS.Multivariate survival analysis showed that the high density of CD45RO+TILs in the stromal region of NSCLC(HR=0.559,95%CI:0.377-0.829,P=0.004)and lung adenocarcinoma(HR=0.352,95%CI:0.193-0.641,P=0.001)were independent prognostic factors for overall survival time(OS).Combined with PD-L1 score of tumor cells in tumor tissues and infiltration score of CD45RO+TILs in all tumor tissues,the patients were divided into 4 groups:patients with PD-L1+/CD45RO+had the longest disease-free survival(DFS)time,and patients with PD-L1+/CD45RO-had the shortest DFS time.Multivariate Cox regression analysis showed that PD-L1+/CD45RO-was an independent prognostic factor for DFS and had a higher risk of poor prognosis compared to the other three groups(HR=2.221,95%CI:1.258-3.919,P=0.006).Conclusion In tumor tissues,the density of CD45RO+TILs,as well as the combination of CD45RO+TILs and PD-L1 in tumor areas,significantly correlated with clinicopathological features and prognosis of NSCLC,which can be used as a new prognosis marker.