CAJ | 학술논문

目的探讨胸腔积液PEMC中CD+4T淋巴细胞分泌或产生IFN-γ的检测对结核性胸膜炎的辅助诊断价值.方法分离40例结核性胸腔积液(结核组)和30例恶性胸腔积液患者(疾病对照组)PEMC,经克隆表达的早期分泌抗原靶6(ESAT-6)和培养滤液蛋白10(CFP-10)融合蛋白(简称"E/C")刺激后,用ELISpot检测分泌IFN-γ细胞的斑点形成细胞(SFC)数量和流式细胞术(FCM)结合胞内细胞因子染色检测产生IFN-γ细胞比率,并比较这2项指标在结核组和疾病对照组间的差异,同时评价2种方法检测IFN-γ对结核性胸膜炎的辅助诊断效能.结果 PEMC经E/C刺激后用ELISpot检测结核组SFC数量为205(125 ～450)SFC/5×104PEMC,疾病对照组为5(2～18)SFC/5×104PEMC,两组间的差异有统计学意义(U=20.00,P<0.01);FCM检测结核组CD+4T淋巴细胞产生IFN-γ细胞比率为3.27%(1.81%～7.34%),疾病对照组为0.12%(0.06%～0.46%),两组间的差异有统计学意义(U=45.00,P<0.01).ELISpot检测PEMC经E/C刺激后分泌IFN-γ的敏感度为92.5%(37/40)、特异度为80.0%(24/30)、阳性预测值为0.86、阴性预测值为0.89、阳性似然比为4.63和阴性似然比为0.09,准确性为87.1%;FCM检测经E/C刺激后产生IFN-γ的敏感度为87.5%(35/40)、特异度为90.0%(27/30)、阳性预测值为0.92、阴性预测值为0.84、阳性似然比为8.75和阴性似然比为0.14,准确性为88.6%.结论经E/C刺激后,用FCM和ELISpot检测CD+4T淋巴细胞产生和分泌IFN-γ细胞的方法对结核性胸膜炎诊断的敏感度和特异度高,且对结核性胸膜炎有辅助诊断价值.
목적 탐토흉강적액PEMC중CD+4T림파세포분비혹산생IFN-γ적검측대결핵성흉막염적보조진단개치.방법 분리40례결핵성흉강적액(결핵조)화30례악성흉강적액환자(질병대조조)PEMC,경극륭표체적조기분비항원파6(ESAT-6)화배양려액단백10(CFP-10)융합단백(간칭"E/C")자격후,용ELISpot검측분비IFN-γ세포적반점형성세포(SFC)수량화류식세포술(FCM)결합포내세포인자염색검측산생IFN-γ세포비솔,병비교저2항지표재결핵조화질병대조조간적차이,동시평개2충방법검측IFN-γ대결핵성흉막염적보조진단효능.결과 PEMC경E/C자격후용ELISpot검측결핵조SFC수량위205(125 ～450)SFC/5×104PEMC,질병대조조위5(2～18)SFC/5×104PEMC,량조간적차이유통계학의의(U=20.00,P<0.01);FCM검측결핵조CD+4T림파세포산생IFN-γ세포비솔위3.27%(1.81%～7.34%),질병대조조위0.12%(0.06%～0.46%),량조간적차이유통계학의의(U=45.00,P<0.01).ELISpot검측PEMC경E/C자격후분비IFN-γ적민감도위92.5%(37/40)、특이도위80.0%(24/30)、양성예측치위0.86、음성예측치위0.89、양성사연비위4.63화음성사연비위0.09,준학성위87.1%;FCM검측경E/C자격후산생IFN-γ적민감도위87.5%(35/40)、특이도위90.0%(27/30)、양성예측치위0.92、음성예측치위0.84、양성사연비위8.75화음성사연비위0.14,준학성위88.6%.결론 경E/C자격후,용FCM화ELISpot검측CD+4T림파세포산생화분비IFN-γ세포적방법대결핵성흉막염진단적민감도화특이도고,차대결핵성흉막염유보조진단개치.
Objective To explore the value of IFN-γ produced or secreted by CD+4 T Lymphocytes from pleural effusion mononuclear cells for the diagnosis of tuberculous pleurisy(plTB).Methods The PEMCs of 40 patients with tuberculous pleural effusion and 30 patients with malignancy pleural effusion were selected as the tuberculosis and disease control groups, then co-cultured with the early secretory antigenic target 6 (ESAT-6) and culture filtered protein 10 (CFP-10) fusion protein (E/C).The numbers of spot forming cells(SFC) secreting IFN-γ were enumerated by ELISpot and the ratios of cells producing IFN-γ were detected by flow cytometry and intracellular cytokine staining.Moreover, the two indicators were compared between tuberculosis and disease control groups to evaluate the 2 methods detecting IFN-γ in the diagnosis of plTB.Results After E/C stimulation, the numbers of SFC were 205(125-450)SFC/5×104 PEMC in tuberculosis group and 5(2-18)SFC/5×104 PEMC in disease control group by ELISpot.The difference between two groups was statistically significant (U= 20.00, P<0.01).The proportion of IFN-γ-secreting CD+4 T lymphocytes was 3.27% (1.81%-7.34%) in tuberculosis group and 0.12% (0.06%-0.46%) in control group detected by FCM. The difference between the two groups was statistically significant (U=45.00, P<0.01).The indicators of ELISpot in detection of IFN-γ which was secreted by PEMC after co-cultured with E/C were as follows: sensitivity 92.5% (37/40), specificity 80.0% (24/30), positive predictive value 0.86, negative predictive value 0.89, positive likelihood ratio 4.63, negative likelihood ratio 0.09 and accuracy 87.1%;and for FCM, they were 87.5% (35/40), 90.0% (27/30), 0.92, 0.84, 8.75 and 0.14, respectively and accuracy 88.6%.Conclusion After E/C stimulation, the assay for IFN-γ-secreting CD+4 T lymphocytes by FCM and ELISpot is highly sensitive and specific for diagnosis of plTB as an auxiliary method.