Email Alert | RSS    帮助

中国防痨杂志 ›› 2021, Vol. 43 ›› Issue (8): 808-812.doi: 10.3969/j.issn.1000-6621.2021.08.010

• 论著 • 上一篇    下一篇

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

王丽, 熊坤龙, 朱长太(), 范琳()   

  1. 201306 上海海洋大学水产与生命学院(王丽);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(熊坤龙、范琳);上海市第六人民医院输血科(朱长太)
  • 收稿日期:2021-04-22 出版日期:2021-08-10 发布日期:2021-07-30
  • 通信作者: 王丽,朱长太,范琳 E-mail:zct101@163.com;fanlinsj@163.com
  • 基金资助:
    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)

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

WANG Li, XIONG Kun-long, ZHU Chang-tai(), FAN Lin()   

  1. *College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306,China
  • Received:2021-04-22 Online:2021-08-10 Published:2021-07-30
  • Contact: WANG Li,ZHU Chang-tai,FAN Lin E-mail:zct101@163.com;fanlinsj@163.com

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%(0.35%,0.91%)],差异均有统计学意义(Z值分别为-2.337、-1.988、-4.455,P值均<0.05)。DS-PTB组CD4-TNF-α分泌水平[0.83%(0.67%,1.22%)]明显高于HD组[0.44%(0.28%,0.71%)]和MDR-PTB组[0.41%(0.28%,0.82%)],差异均有统计学意义(Z值分别为-3.903、-2.919,P值均<0.05)。结论 MDR-PTB患者存在T淋巴细胞功能耗竭,阻碍了T淋巴细胞免疫效应的表达。

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

Abstract:

Objective 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-γ of DS-PTB (0.92% (0.59%, 2.02%)) was significantly higher than that of HD group (0.65% (0.38%, 0.82%)), CD8-IFN-γ of 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%)), the differences were statistically significant (Z=-2.337, -1.988,-4.455, all P values <0.05). TNF-α level of DS-PTB (0.83% (0.67%, 1.22%)) was higher than that of HD (0.44% (0.28%, 0.71%)) and MDR-PTB (0.41% (0.28%, 0.82%)), the differences were statistically significant (Z=-3.903, -2.919,all P values <0.05). Conclusion T lymphocyte exhaustion exists in patients with MDR-PTB, T lymphocyte immune effect was inhibited by cells exhaustion.

Key words: Tuberculosis,multidrug-resistant, Immunologic tests, T-lymphocyte subsets, Apoptosis, Evaluation studies