结核分枝杆菌通过CD14^(+)细胞诱导固有淋巴细胞亚群分化失衡的机制探讨  

Mycobacterium tuberculosis induces differentiation imbalance of innate lymphoid cells via CD14^(+)monocytes/macrophages

作　　者：张苏 欧敏 周雪峰 莫雨平[3] 曹廷智[3] 张国良[1] Zhang Su;Ou Min;Zhou Xuefeng;Mo Yuping;Cao Tingzhi;Zhang Guoliang(National Clinical Research Center for Infectious Diseases,Shenzhen Third People′s Hospital,Shenzhen 518112,China;School of Basic Medical Sciences,Guangzhou Medical University,Guangzhou 510000,China;Traditional Chinese Medicine Department,Shenzhen Third People′s Hospital,Shenzhen 518112,China)

机构地区：[1]深圳市第三人民医院国家感染性疾病临床医学研究中心,深圳518112 [2]广州医科大学基础医学院,广州510000 [3]深圳市第三人民医院中医科,深圳518112

出　　处：《中华传染病杂志》2024年第10期608-617,共10页Chinese Journal of Infectious Diseases

基　　金：广东省科技计划项目(2020B1111170014);深圳市科技计划项目(KCXFZ20211020163545004、RCJC20221008092726022);深圳市"医疗卫生三名工程"项目(SZZYSM202311009)。

摘　　要：目的分析不同结核分枝杆菌(MTB)感染状态人群外周血中固有淋巴细胞(ILC)各亚群的比例,探讨MTB感染过程中调控ILC各亚群分化的分子机制。方法纳入深圳市第三人民医院2022年3月至10月收治的活动性肺结核患者31例、肺结核治愈患者17例,同时从体检科招募健康对照者30名,采集所有受试者的外周血并提取外周血单个核细胞(PBMC),采用流式细胞术分析不同人群PBMC中ILC、ILC1、ILC2、ILC3占淋巴细胞的比例。分离18名健康对照者PBMC,体外使用MTB标准株H37Rv裂解物或卡介苗(BCG)活菌诱导PBMC,采用流式细胞术分析ILC各亚群分化情况。采用磁珠分选技术分离29名健康对照者PBMC中的CD14^(+)细胞,分为对照组、CD14-组和CD14^(+)回补组,其中CD14^(+)回补组通过Transwell小室向CD14-PBMC中回补CD14^(+)细胞,通过H37Rv裂解物体外诱导48 h,采用流式细胞术分析3组细胞中ILC各亚群分化情况。统计学分析采用曼-惠特尼U检验、Kruskal-Wallis检验、威尔科克森符号秩检验。结果健康对照者、活动性肺结核、肺结核治愈人群中ILC占淋巴细胞的比例差异无统计学意义(H=0.07,P=0.965)。与健康对照组比较,活动性肺结核患者外周血中ILC1占淋巴细胞的比例较低(U=271.00),而其ILC2占淋巴细胞的比例较高(U=299.00);肺结核治愈患者ILC1占淋巴细胞的比例低于健康对照组(U=123.00),而其ILC3的比例分别高于活动性肺结核组和健康对照组(U=78.00、102.50),差异均有统计学意义(均P<0.05)。PBMC经H37Rv裂解物或BCG活菌诱导后,两组细胞的ILC(W=-116.00、-145.00)和ILC2(W=-149.00、-155.00)占淋巴细胞的比例均低于对照组,差异均有统计学意义(均P<0.05);ILC1占淋巴细胞的比例在经BCG活菌诱导后降低(W=-121.00,P=0.007),其在经H37Rv裂解物诱导后与对照组差异无统计学意义(W=-67.00,P=0.154)。对照组、CD14-组和CD14^(+)回补组细胞经H37Rv裂解物诱导后,三组细胞ILC1占淋巴�ObjectiveTo analyze the proportions of innate lymphoid cells(ILCs)subgroups during the process of Mycobacterium tuberculosis(MTB)infection,and to explore the molecular mechanism regulating the differentiation of ILCs during MTB infection.MethodsFrom March to October 2022,31 patients with active pulmonary tuberculosis(ATB)and 17 patients who had recovered from pulmonary tuberculosis were enrolled from Shenzhen Third People′s Hospital.Additionally,30 healthy controls were recruited from the physical examination department.Peripheral blood mononuclear cells(PBMCs)were extracted from all subjects,and the proportions of ILC,ILC1,ILC2 and ILC3 in lymphocytes of PBMCs from different populations were analyzed using flow cytometry.PBMCs from 18 healthy controls were induced in vitro with MTB H37Rv lysate or live Bacillus Calmette-Guérin(BCG)bacteria,and the differentiation of ILC subgroups was analyzed using flow cytometry.CD14^(+)cells from PBMCs of 29 healthy controls were isolated using magnetic bead sorting technology,and the cells were divided into three groups,including control group,CD14-group,and CD14^(+)complement group.The CD14^(+)complement group was supplemented with CD14^(+)cells into CD14-PBMCs through a Transwell chamber,and induced in vitro with H37Rv lysate.The differentiation of ILC subgroups was analyzed using flow cytometry.Statistical analyses were performed using Mann-Whitney U test,Kruskal-Wallis test,and Wilcoxon signed-rank test.ResultsThe proportions of ILCs in lymphocytes in healthy controls,ATB and recovered tuberculosis groups showed no statistically significant differences(H=0.07,P=0.965).The proportion of ILC1 in lymphocytes in the peripheral blood of patients with ATB was lower than that in the healthy control group(U=271.00),and the proportion of ILC2 was higher than that in the healthy control group(U=299.00).The proportion of ILC1 in the peripheral blood of recovered tuberculosis patients was lower than that in the healthy control group(U=123.00),and the proportion of ILC3 in the reco

关 键 词：结核分枝杆菌 固有淋巴细胞 活动性肺结核 单核巨噬细胞 

分 类 号：R521[医药卫生—内科学]