胸腹水、脑脊液中结核分枝杆菌RD1基因编码抗原刺激后释放γ干扰素的特异性T细胞检测

Detection of Specific Interferon-γ-secreting T Cell Response to Mycobacterium Tuberculosis RD1-encoded Antigens in Pleural Effusions,Ascites,and Cerebrospinal Fluid

目的初步探讨胸腹水、脑脊液中结核分枝杆菌RD1基因编码抗原刺激后释放γ干扰素的特异性T细胞检测在诊断结核感染中的应用。方法应用酶联免疫斑点技术检测6kD早期分泌靶向抗原和10kD培养滤过蛋白肽段库刺激后外周血、胸腹水、脑脊液单个核细胞中释放γ干扰素的特异性T细胞。结果静脉血和胸腹水、脑脊液中单个核细胞γ干扰素释放反应有助于诊断结核感染,结果阳性的患者最终均诊断为活动性结核病。结果阳性的胸、腹水斑点形成细胞数分别是静脉血的6.4和31.9倍。结论在胸腹水、脑脊液中检测结核分枝杆菌RD1基因编码抗原刺激后释放γ干扰素的特异性T细胞,可能为诊断结核感染提供新的途径。

Objective To detect Mycobacterium tuberculosis RD1-encoded antigens-specific,interferon-gamma（INF-γ）-secreting T cells in pleural effusions,ascites,and cerebrospinal fluid. Method The early secretory antigenic target-6 （ESAT-6） and culture filtrate protein-10 （CFP-10） peptides-specific T cells in peripheral blood mononuclear cell （MC）,ascites MC,pleural effusions MC,and cerebrospinal fluid MC were detected using enzyme-linked immunospot assay （ELISPOT） for INF-γ. Results ESAT-6 or CFP-10 peptides-specific, INF-γ-secreting T cells were detected in peripheral blood, ascites, pleural effusions, and cerebrospinal fluid, which marked the presence of tuberculosis infection. Patients with positive ELISPOT results of INF-γ-release assay were all diagnosed as active tuberculosis. Spot forming cells in ascites and pleural effusions were much higher than those in peripheral blood （ up to 6.4 and 31.9 times）. Conclusion Detection of RDl-encoded antigens-specific, INF-γ-secreting T cells in pleural effusions, ascites, and cerebrospinal fluid provides a new way to diagnose tuberculosis infection.