CAJ | 학술논문

目的应用流式细胞术检测胸腔积液单个核细胞经早期分泌抗原靶蛋白-6(ESAT-6)/培养滤过蛋白-10(CFP-10)融合蛋白刺激后Th1、Th2细胞百分率,探讨结核性胸腔积液局部抗原特异性Th1、Th2反应特点及对结核性胸膜炎的诊断价值.方法 2008年9月至2009年3月期间在北京胸科医院收治的结核性胸腔积液40例为结核组,其中男30例,女10例；年龄16 ～53岁,平均(29±12)岁；同期收治的恶性胸腔积液患者30例为肿瘤组,其中男23例,女7例；年龄35 ～ 65岁,平均(47±15)岁.分离胸腔积液单个核细胞并冻存,经细胞复苏、与抗原共培养(同时设立阴性对照和阳性对照),应用流式细胞内细胞因子染色技术检测抗原特异性Th1、Th2细胞百分率.两组间正态分布数据采用￡检验,非正态分布数据采用Wilcoxon检验.结果结核组单个核细胞经抗原刺激后Th1百分率及Th 1/Th2比值的中位数(四分位间距)分别为3.06％(1.59％～6.92％)和17.00(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)]和肿瘤组[0.12％ (0.05％～0.39％)和1.05(0.25 ～2.52)],差异均有统计学意义(Z值为-6.624 ～-3.314,均P＜0.01),Th2细胞百分率(0.22±0.19)％显著高于阴性对照(0.10±0.08)％,差异有统计学意义(t=4.108,P＜0.01)；与肿瘤组[(0.15±0.02)％]比较,差异无统计学意义(t=1.954,P＞0.05).Th1诊断结核性胸腔积液的曲线下面积(0.937)和敏感度(85.4％)显著高于Th 1/Th2(0.883和81.5％),两种方法的特异度一致(均为90.6％).结论结核性胸腔积液中ESAT-6/CFP-10融合蛋白特异性Th1、Th2反应为以Th1占明显优势的Th1/Th2混合反应,Th1和Th1/Th2有望成为流式细胞内细胞因子染色技术鉴别结核性与恶性胸腔积液的指标.
목적 응용류식세포술검측흉강적액단개핵세포경조기분비항원파단백-6(ESAT-6)/배양려과단백-10(CFP-10)융합단백자격후Th1、Th2세포백분솔,탐토결핵성흉강적액국부항원특이성Th1、Th2반응특점급대결핵성흉막염적진단개치.방법 2008년9월지2009년3월기간재북경흉과의원수치적결핵성흉강적액40례위결핵조,기중남30례,녀10례；년령16 ～53세,평균(29±12)세；동기수치적악성흉강적액환자30례위종류조,기중남23례,녀7례；년령35 ～ 65세,평균(47±15)세.분리흉강적액단개핵세포병동존,경세포복소、여항원공배양(동시설립음성대조화양성대조),응용류식세포내세포인자염색기술검측항원특이성Th1、Th2세포백분솔.량조간정태분포수거채용￡검험,비정태분포수거채용Wilcoxon검험.결과 결핵조단개핵세포경항원자격후Th1백분솔급Th 1/Th2비치적중위수(사분위간거)분별위3.06％(1.59％～6.92％)화17.00(7.38 ～35.53),현저고우음성대조[0.38％(0.02％～1.80％)화3.59 (0.49～25.09)]화종류조[0.12％ (0.05％～0.39％)화1.05(0.25 ～2.52)],차이균유통계학의의(Z치위-6.624 ～-3.314,균P＜0.01),Th2세포백분솔(0.22±0.19)％현저고우음성대조(0.10±0.08)％,차이유통계학의의(t=4.108,P＜0.01)；여종류조[(0.15±0.02)％]비교,차이무통계학의의(t=1.954,P＞0.05).Th1진단결핵성흉강적액적곡선하면적(0.937)화민감도(85.4％)현저고우Th 1/Th2(0.883화81.5％),량충방법적특이도일치(균위90.6％).결론 결핵성흉강적액중ESAT-6/CFP-10융합단백특이성Th1、Th2반응위이Th1점명현우세적Th1/Th2혼합반응,Th1화Th1/Th2유망성위류식세포내세포인자염색기술감별결핵성여악성흉강적액적지표.
Objective 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.Methods 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.Results 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.Conclusions The feature of ESAT-6/CFP-10 fusion protein-specific Th1 and Th2 response in tuberculous pleurisy was a mixed reaction of Thl and Th2 with Th1 predominance.Th1 percentage and Th1/Th2 ratio could be diagnostic indexes for identifying tuberculous from malignant pleural effusions.