Ge Qi-ping, He Zheng-yi, Gao Wei-wei, DU Feng-jiao, Wei Pan-jian, Jia Hong-yan, Ma Yu, Zhang Zong-de
Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis & Thoracic Tumor Research Institute, Beijing 101149, China.
Zhonghua Jie He He Hu Xi Za Zhi. 2013 Jun;36(6):406-10.
To detect the Th1 and Th2 cell percentage in pleural effusion mononuclear cells (PEMCs) stimulated by early secretory antigenic target protein-6 (ESAT-6)/culture filtrate protein-10 (CFP-10) fusion protein (E/C) with flow cytometry (FCM), and therefore to explore the local antigen specific Th1 and Th2 response and its diagnostic value in tuberculous pleuritis.
Forty patients with tuberculous pleural effusion and 30 patients with malignant pleural effusion were included in this study from Sep.2008 to Mar.2009. PEMCs were isolated and cryopreserved. After resuscitation, the cells were cultured with E/C (simultaneously with positive control and negative control), and antigen-specific Th1 and Th2 cells were detected with intracellular cytokine staining of FCM. Normal distribution data using t test, abnormal distribution data using Wilcoxon test.
In the TB group,the medians (quartile range) of Th1 cells and Th1/Th2 ratio among PEMCs stimulated by ESAT-6/CFP-10 fusion protein were 3.06% (1.59%-6.92%) and 17 (7.38-35.53), significantly higher than those of the negative control [0.38% (0.02%-1.80%) and 3.59 (0.49-25.09)], the differences being statistically significant (Z = -5.345 and 3.314, P < 0.01). The percentage of Th2 cells [(0.22 ± 0.19)%] was also increased compared with that of the negative control [(0.10 ± 0.08)%], the difference being statistically significant (t = 4.108, P < 0.01). In the malignant effusion group, the medians (quartile range) of Th1 percentage and Th1/Th2 ratio were 0.12% (0.05%-0.39%) and 1.05 (0.25-2.52), which were significantly different as compared with those of the TB group (Z = -6.624 and -5.536, P < 0.01). The Th2 percentage in the 2 groups were (0.22 ± 0.19)% and (0.15 ± 0.02)%, respectively (t = 1.954, P > 0.05). The receiver operating characteristic curve indicated that the area under the curve (AUC), sensitivity, and specificity were 0.937, 85.4%, and 90.6% respectively for Th1 to diagnose tuberculous pleurisy. For Th1/Th2, the AUC, sensitivity, and specificity were 0.883, 81.5%, and 90.6% respectively.
The feature of ESAT-6/CFP-10 fusion protein-specific Th1 and Th2 response in tuberculous pleurisy was a mixed reaction of Th1 and Th2 with Th1 predominance. Th1 percentage and Th1/Th2 ratio could be diagnostic indexes for identifying tuberculous from malignant pleural effusions.
采用流式细胞术(FCM)检测早期分泌性抗原靶蛋白6(ESAT-6)/培养滤液蛋白10(CFP-10)融合蛋白(E/C)刺激的胸腔积液单个核细胞(PEMCs)中Th1和Th2细胞百分比,以探讨结核性胸膜炎局部抗原特异性Th1和Th2反应及其诊断价值。
选取2008年9月至2009年3月间40例结核性胸腔积液患者和30例恶性胸腔积液患者。分离并冻存PEMCs。复苏后,将细胞与E/C共同培养(同时设阳性对照和阴性对照),采用FCM细胞内细胞因子染色法检测抗原特异性Th1和Th2细胞。正态分布数据采用t检验,非正态分布数据采用Wilcoxon检验。
在结核组中,ESAT-6/CFP-10融合蛋白刺激的PEMCs中Th1细胞中位数(四分位间距)及Th1/Th2比值分别为3.06%(1.59%-6.92%)和17(7.38-35.53),显著高于阴性对照[0.38%(0.02%-1.80%)和3.59(0.49-25.09)],差异有统计学意义(Z=-5.345和3.314,P<0.01)。Th2细胞百分比[(0.22±0.19)%]较阴性对照[(0.10±0.08)%]也有所升高,差异有统计学意义(t=4.108,P<0.01)。在恶性胸腔积液组中,Th1百分比及Th1/Th2比值中位数(四分位间距)分别为0.12%(0.05%-0.39%)和1.05(0.25-2.52),与结核组相比差异有统计学意义(Z=-6.624和-5.536,P<0.01)。两组Th2百分比分别为(0.22±0.19)%和(0.15±0.02)%(t=1.954,P>0.05)。受试者工作特征曲线表明,Th1诊断结核性胸膜炎的曲线下面积(AUC)、灵敏度和特异度分别为0.937、85.4%和90.6%。对于Th1/Th2,AUC、灵敏度和特异度分别为0.883、81.5%和90.6%。
结核性胸膜炎中ESAT-6/CFP-10融合蛋白特异性Th1和Th2反应的特点是Th1和Th2的混合反应,以Th1为主。Th1百分比及Th1/Th2比值可作为鉴别结核性与恶性胸腔积液的诊断指标。