Barnes P F, Lu S, Abrams J S, Wang E, Yamamura M, Modlin R L
Department of Medicine, University of Southern California School of Medicine, Los Angeles 90033.
Infect Immun. 1993 Aug;61(8):3482-9. doi: 10.1128/iai.61.8.3482-3489.1993.
Clinical and immunologic evidence suggests that tuberculous pleuritis provides a model to understand protective immune mechanisms against Mycobacterium tuberculosis. We therefore evaluated the pattern of cytokine mRNA expression and cytokine production in pleural fluid and blood of patients with tuberculous pleuritis. RNA was extracted from mononuclear cells, reverse transcribed to cDNA, and amplified by polymerase chain reaction (PCR). After normalization for T-cell cDNA, cDNA from pleural fluid cells and peripheral blood mononuclear cells (PBMC) was amplified with cytokine-specific primers. PCR product was quantified by Southern blot. For the Th1 cytokines gamma interferon (IFN-gamma) and interleukin-2 (IL-2), PCR product was greater in pleural fluid than in blood, whereas PCR product for the Th2 cytokine IL-4 was decreased in pleural fluid compared with blood. Concentrations of IFN-gamma were elevated in pleural fluid compared with serum, but IL-2, IL-4, and IL-5 were not detectable. Mean concentrations of IFN-gamma and IL-2 in supernatants of M. tuberculosis-stimulated pleural fluid cells were significantly greater than corresponding concentrations in supernatants of stimulated PBMC. In situ hybridization showed that increased IFN-gamma production by pleural fluid cells was associated with a 20- to 60-fold increase in the frequency of antigen-reactive IFN-gamma-mRNA-expressing cells. Because IL-10 can be produced by T cells and macrophages, pleural fluid cells and PBMC were normalized for beta-actin cDNA content and then amplified by PCR with IL-10-specific primers. IL-10 mRNA was greater in pleural fluid cells than in PBMC and was expressed predominantly by macrophages. IL-10 concentrations were elevated in pleural fluid versus serum. These data provide strong evidence for compartmentalization of Th1 cytokines and IL-10 at the site of disease in humans with a resistant immune response to mycobacterial infection.
临床和免疫学证据表明，结核性胸膜炎为理解针对结核分枝杆菌的保护性免疫机制提供了一个模型。因此，我们评估了结核性胸膜炎患者胸腔积液和血液中细胞因子mRNA表达模式及细胞因子产生情况。从单核细胞中提取RNA，反转录为cDNA，并用聚合酶链反应（PCR）进行扩增。在对T细胞cDNA进行标准化后，用细胞因子特异性引物对胸腔积液细胞和外周血单核细胞（PBMC）的cDNA进行扩增。通过Southern印迹对PCR产物进行定量。对于Th1细胞因子γ干扰素（IFN-γ）和白细胞介素-2（IL-2），胸腔积液中的PCR产物比血液中的多，而与血液相比，Th2细胞因子IL-4的PCR产物在胸腔积液中减少。与血清相比，胸腔积液中IFN-γ的浓度升高，但未检测到IL-2、IL-4和IL-5。结核分枝杆菌刺激的胸腔积液细胞上清液中IFN-γ和IL-2的平均浓度显著高于刺激的PBMC上清液中的相应浓度。原位杂交显示，胸腔积液细胞中IFN-γ产生增加与表达抗原反应性IFN-γ-mRNA的细胞频率增加20至60倍相关。由于IL-10可由T细胞和巨噬细胞产生，因此对胸腔积液细胞和PBMC的β-肌动蛋白cDNA含量进行标准化，然后用IL-10特异性引物通过PCR进行扩增。胸腔积液细胞中的IL-10 mRNA比PBMC中的多，且主要由巨噬细胞表达。胸腔积液中的IL-10浓度相对于血清升高。这些数据为在对分枝杆菌感染具有抗性免疫反应的人类疾病部位Th1细胞因子和IL-10的分隔提供了有力证据。
