T淋巴细胞耗竭在耐多药肺结核患者免疫表达中的初步研究  

T lymphocyte exhaustion on the expression of immune effect in patients with multidrug-resistant pulmonary tuberculosis

作　　者：王丽 熊坤龙 朱长太[3] 范琳[2] WANG Li;XIONG Kun-long;ZHU Chang-tai;FAN Lin(College of Fisheries and Life Sciences,Shanghai Ocean University,Shanghai 201306,China;不详)

机构地区：[1]上海海洋大学水产与生命学院,201306 [2]同济大学附属上海市肺科医院结核科,上海市感染性疾病(结核病)医学临床研究中心 [3]上海市第六人民医院输血科

出　　处：《中国防痨杂志》2021年第8期808-812,共5页Chinese Journal of Antituberculosis

基　　金：上海市科委自然基金(20ZR1446700);上海市浦东新区科技发展基金(PKJ-2020-Y12)。

摘　　要：目的探讨T淋巴细胞耗竭在耐多药肺结核(multidrug-resistant pulmonary tuberculosis,MDR-PTB)患者外周血分离流式细胞术检测后免疫表达中的特征。方法采用前瞻性研究的方法,选取2020年1—8月同济大学附属上海市肺科医院住院治疗的58例肺结核患者作为研究对象,其中,MDR-PTB患者20例(MDR-PTB组),药物敏感性肺结核(drug susceptible pulmonary tuberculosis,DS-PTB)患者38例(DS-PTB组);同期纳入健康志愿者20名,作为健康对照组(HD组)。分别采集各组研究对象肘部静脉血各10ml并抗凝处理,分离其外周血单个核细胞(peripheral blood mononuclear,PBMC),利用流式细胞术检测并比较其结核分枝杆菌(Mycobacterium tuberculosis,MTB)抗原特异性CD4+和CD8+T淋巴细胞表面程序性死亡受体-1(PD-1)、T细胞免疫球蛋白黏蛋白分子-3(Tim-3)和淋巴细胞活化基因-3(Lag-3)等抑制性受体的表达水平,以及胞内细胞因子白细胞介素-2(IL-2)、γ-干扰素(IFN-γ)和肿瘤坏死因子-α(TNF-α)的分泌水平。结果MDR-PTB组CD4-PD-1、CD8-PD-1、CD4-Tim-3和CD8-Tim-3的表达水平[中位数(四分位数)][分别为24.50%(21.58%,26.00%)、19.95%(16.10%,21.65%)、3.75%(3.20%,4.68%)和12.65%(10.65%,14.10%)]均明显高于HD组[分别为13.10%(11.80%,17.80%)、12.55%(9.61%,18.18%)、0.67%(0.46%,2.41%)和3.78%(2.35%,6.55%)]和DS-PTB组[分别为14.50%(11.90%,20.00%)、13.20%(8.61%,18.23%)、1.12%(0.71%,3.36%)和3.59%(2.10%,7.52%)],差异均有统计学意义(Z值分别为-4.009、-4.159,-3.027、-3.284,-3.565、-3.967,-5.568、-3.261,P值均<0.05)。MDR-PTB组CD4-IL-2和CD8-IL-2分泌水平[分别为1.30%(0.73%,2.71%)和2.27%(1.03%,3.11%)]均明显高于HD组[分别为0.67%(0.44%,1.10%)和0.59%(0.39%,0.91%)],差异均有统计学意义(Z=-2.670、-3.917,P值均<0.05)。DS-PTB组CD4-IFN-γ分泌水平[0.92%(0.59%,2.02%)]明显高于HD组[0.65%(0.38%,0.82%)],MDR-PTB组和DS-PTB组CD8-IFN-γ分泌水平[分别为1.16%(0.53%,2.03%)和1.80%(0.97%,3.04%)]均明显高于HD组[0.69%(0.3Objective To explore the characteristics of T lymphocyte exhaustion in immune responses from peripheral blood of patients with multidrug-resistant pulmonary tuberculosis(MDR-PTB)by flow cytometry.Methods Fifty-eight patients with pulmonary tuberculosis hospitalized in Shanghai Pulmonary Hospital affilicated to Tongji University were prospectively included from January 2020 to August in 2020,including 20 patients with MDR-PTB,38 patients with drug susceptible pulmonary tuberculosis(DS-PTB),and 20 healthy donors as control(HD).The elbow vein anticoagulant blood(10ml)of each patient was collected,peripheral blood mononuclear(PBMC)cells were isolated,MTB-specific programmed death-ligand-1(PD-1),T lymphocyte immunoglobulin and mucin-domain-containing molecules-3(Tim-3)and lymphocyte activation gene-3(Lag-3)on CD4+and CD8+T lymphocytes and intracellular cytokines of interleukin-2(IL-2),interferon gamma(IFN-γ)and tumor necrosis factor-α(TNF-α)were detected by flow cytometry.Results The expression of CD4-PD-1,CD8-PD-1,CD4-Tim-3 and CD8-Tim-3 in MDR-PTB(median(quartile))(respectively were 24.50%(21.58%,26.00%),19.95%(16.10%,21.65%),3.75%(3.20%,4.68%)and 12.65%(10.65%,14.10%)),which were significantly higher than those of HD(respectively were 13.10%(11.80%,17.80%),12.55%(9.61%,18.18%),0.67%(0.46%,2.41%)and 3.78%(2.35%,6.55%))and DS-PTB(respectively were 14.50%(11.90%,20.00%),13.20%(8.61%,18.23%),1.12%(0.71%,3.36%)and 3.59%(2.10%,7.52%)),the differences were statistically significant(Z=-4.009,-4.159,-3.027,-3.284,-3.565,-3.967,-5.568,-3.261,all P values<0.05).CD4-IL-2 and CD8-IL-2 levels of MDR-TB(1.30%(0.73%,2.71%),2.27%(1.03%,3.11%))were higher than those of HD(0.67%(0.44%,1.10%),0.59%(0.39%,0.91%)),the differences were statistically significant(Z=-2.670,-3.917,all P values<0.05).CD4-IFN-γof DS-PTB(0.92%(0.59%,2.02%))was significantly higher than that of HD group(0.65%(0.38%,0.82%)),CD8-IFN-γof MDR-PTB and DS-PTB(1.16%(0.53%,2.03%)and 1.80%(0.97%,3.04%))were significantly higher than that of HD group(0.69%(0.35%,0.91

关 键 词：结核 抗多种药物性 免疫学试验 T淋巴细胞亚群 细胞凋亡 评价研究 

分 类 号：R521[医药卫生—内科学]