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目的:基于弥漫大B细胞淋巴瘤（DLBCL）基因芯片表达数据库，分析肿瘤相关巨噬细胞及亚型浸润情况，探讨其与患者预后的相关性。方法:数据来源于PubMed基因表达数据库（GEO）中的DLBCL患者微阵列（Affymetrix U133 plus 2.0）数据库（编号GSE10846）。该数据库共包含414例患者，其中306例有完整的临床、细胞起源分型（COO分型）及治疗随访信息。通过基于评估RNA转录本相对占比进行细胞类型识别（CIBERSORT）的在线计算机程序进行数据分析；从生成的分析结果文件中识别出GSE10846数据库中所有患者包含各亚型巨噬细胞在内的免疫细胞在微环境中所有可识别免疫细胞中的比例。以巨噬细胞亚群占微环境所有免疫细胞比例中位值为临界值：≥临界值为高浸润。基于GSE10846数据库中所有414例DLBCL患者的myc、bcl-2、程序性死亡受体配体1（PD-L1）及程序性死亡受体配体2（PD-L2）基因表达量的中位值为临界值，≥临界值者为高表达。分析各型及总体巨噬细胞水平与临床因素、一些基因表达及生存的相关性，采用Cox比例风险模型对DLBCL患者预后进行多因素分析。采用R 4.0.4中surv_miner包中的surv_cutpoint函数确定各项巨噬细胞亚群占微环境所有免疫细胞比例的最佳临界值，<最佳临界值为统计学低浸润，≥最佳临界值为统计学高浸润。结果:经CIBERSORT分析，414例DLBCL患者中识别出肿瘤微环境中M0型巨噬细胞[15.00%（0~44.41%）]、M1型巨噬细胞[7.46%（0~23.00%）]、M2型巨噬细胞[6.28%（0~43.35%）]。具有完整的临床和随访资料的306例患者中，M0、M1、M2型和总巨噬细胞高浸润患者分别有155例（50.7%）、152例（49.7%）、156例（51.0%）、152例（49.7%），M0型巨噬细胞高浸润与COO分型为生发中心B细胞（GCB）型、PD-L1基因高表达及无myc、bcl-2 RNA水平双高表达（R-DEL）有关（均n P<0.05），M1型巨噬细胞高浸润与女性、PD-L1基因高表达、PD-L2基因高表达有关（均n P<0.05），M2型巨噬细胞高浸润与COO分型为GCB型、PD-L2基因高表达有关（均n P<0.05），总巨噬细胞高浸润与女性、COO分型为GCB型、PD-L1基因高表达、PD-L2基因高表达、无R-DEL有关（均n P<0.05）。PD-L1基因高表达与M0、M1型及总巨噬细胞高浸润相关（均n P<0.01），PD-L2基因高表达与M1、M2型巨噬细胞及总巨噬细胞高浸润相关（均n P<0.01）。M0型巨噬细胞高浸润组总生存（OS）较低浸润组好（n P=0.002）；M2型巨噬细胞低浸润组OS较高浸润组好（n P=0.019）。R-DEL组OS较无R-DEL组差（n P=0.001）。国际预后指数（IPI）评分低（0~2分）、COO分型为GCB型、治疗中采用利妥昔单抗的患者OS更好（均n P<0.01）。多因素Cox回归分析结果显示，年龄≥60岁、COO分型为非GCB分型、治疗方案中不含利妥昔单抗、M0型巨噬细胞低浸润、M2型巨噬细胞高浸润为DLBCL患者OS的独立不良影响因素（均n P<0.05）。M0型巨噬细胞最佳临界值为4.3%，M2型巨噬细胞最佳临界值为4.8%；M0型巨噬细胞统计学低浸润组OS更差（n P<0.001），M2型巨噬细胞统计学高浸润组OS更差（n P=0.001）。n 结论:DLBCL肿瘤微环境的免疫细胞中肿瘤相关巨噬细胞含量最高。M2型巨噬细胞高浸润患者预后较差，而M0型巨噬细胞高浸润患者预后较好。“,”Objective:To analyze the infiltration of tumor-associated macrophages and their subtypes, and to investigate their association with prognosis of patients with diffuse large B-cell lymphoma (DLBCL) based on the gene chip expression database.Methods:The data were retrieved from microarray (Affymetrix U133 plus 2.0) database (No:GSE10846) of DLBCL patients in PubMed gene expression omnibus (GEO). The database included 414 DLBCL patients, among which 306 cases had complete clinical, cell of origin phenotype (COO subtyping), treatment and follow-up information. The data analysis was performed on the online computer program which could identify the cell-type (CIBERSORT) by estimating relative percentage of RNA transcripts. From the returned result file, the percentage of immune cells including macrophages subtypes of all cases in all identifiable immune cells in the microenvironment was identified in GSE10846 database. Taking the median percentage of macrophages subsets in all types of immune cells as cut-off value; ≥ cut-off value was high infiltration and < cut-off value was low infiltration. The median value of gene RNA expression level of myc, bcl-2, programmed death ligand-1 (PD-L1) and programmed death ligand-2 (PD-L2) of 414 DLBCL patients in the GSE10846 database was treated as the cut-off value; ≥ cut-off value was the high expression and < cut-off value was the low expression. The correlation of the expression levels of all subsets and total macrophages with clinical factors, gene expression, survival was analyzed; Cox proportional hazard model was used to make multivariate analysis of the prognosis for DLBCL patients. surv_cutpoint function of surv_miner package in R 4.0.4 software was used for the optimal cut-off value of the percentage of macrophages subsets in all immune cells in the microenvironment; the result less than the optimal cut-off value was statistically low infiltration and the result greater than or equal to the optimal cut-off value was statistically high infiltration.Results:CIBERSORT analysis showed that M0 macrophages [15.00% (0-44.41%)], M1 macrophages [7.46% (0-23.00%)] and M2 macrophages [6.28% (0-43.35%)] in the tumor microenvironment were identified in all 414 DLBCL cases. Among 306 patients with complete clinical and follow-up data, there were 155 cases (50.7%), 152 cases (49.7%), 156 cases (51.0%), 152 cases (49.7%), respectively in high infiltration patients with M0, M1, M2 and total macrophages; the high infiltration of M0 macrophages was correlated with COO subtyping germinal center B-cell (GCB) type and the high expression of PD-L1 gene, the absence of myc and bcl-2 double high expression at RNA level (R-DEL) (alln P < 0.05); the high infiltration of M1 macrophages was correlated with female, the high expression of PD-L1 gene and PD-L2 gene (all n P < 0.05); the high infiltration of M2 macrophages was correlated with COO subtyping GCB type, the high expression of PD-L2 gene (all n P < 0.05); the high infiltration of total macrophages was correlated with female, COO subtyping GCB type, the high expression of PD-L1 gene and PD-L2 gene, the absence of R-DEL (all n P < 0.05).The high expression of PD-L1 gene was associated with high infiltration of M0, M1 and total macrophages (all n P < 0.01), and high PD-L2 gene expression was correlated with high infiltration of M1, M2 and total macrophages (all n P < 0.01). The overall survival (OS) of M0 macrophage high infiltration group was better than that of the lower infiltration group ( n P = 0.002); the OS of M2 macrophage low infiltration group was better than that of the high infiltration group (n P = 0.019). The OS of R-DEL group was worse than that of R-DEL absent group (n P = 0.001). The patients with low international prognostic index (IPI) score (0-2), COO subtyping GCB type, and treatment with rituximab had better OS (all n P < 0.01). Multivariate Cox regression analysis showed that 60 years or above, COO subtyping non-GCB type, treatment without rituximab, M0 macrophage low infiltration, M2 macrophage high infiltration were all independent adverse prognostic factors for OS of DLBCL patients (all n P < 0.05). The optimal cut-off value for M0 macrophages was 4.3%, and the optimal cut-off value for M2 macrophages was 4.8%, and the OS in the group with statistically low infiltration of M0 macrophage was worse ( n P < 0.001), and so was the OS in the group with statistically high infiltration of M2 macrophage ( n P = 0.001).n Conclusions:Tumor-associated macrophage is confirmed as the most abundant immune cells in the tumor microenvironment of DLBCL. Patients with high infiltration of M2 macrophage have poor prognosis, while high infiltration of M0 macrophage indicates a better prognosis.