Comparison of Intracellular Cytokine Flow Cytometry and an Enzyme Immunoassay for Evaluation of Cellular Immune Response to Active Tuberculosis
Comparison of Intracellular Cytokine Flow Cytometry and an Enzyme Immunoassay for Evaluation of Cellular Immune Response to Active Tuberculosis
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DOI：
10.1128/cvi.00159-08
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发表时间：
2009-03-01
影响因子：
--
通讯作者：
Kam, Kai Man
中科院分区：
文献类型：
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作者：
Leung, Wai Lin;Law, Kai Leung;Kam, Kai Man
描述了一项前瞻性横断面盲法研究，对象为 28 名在结核病和胸部诊所就诊的疑似活动性结核病 (TB) 患者（21 名男性和 7 名女性患者；平均年龄 44 岁）。采集血液进行免疫细胞计数、全血酶联免疫吸附测定（ELISA），通过QuantiFERON-TB Gold（QFT-G）测定检测γ干扰素（IFN-γ），以及细胞内细胞因子流式细胞术（ICC）分析；同时抽取痰液进行结核杆菌细菌学培养。十二名健康受试者作为对照。比较了 QFT-G 和 ICC 检测活动性结核病的性能特征。在活动性结核病患者中，我们发现 (i) 外周 CD4(+) 和 CD8(+) T 细胞计数正常至轻微升高，但 NK 细胞数量显着减少； (ii) CD4(+) T细胞是产生IFN-γ（一种1型细胞因子）的主要细胞类型； (iii) 小比例的 CD8(+) T 细胞也已准备好产生 IFN-γ； (iv) 2 型细胞因子白细胞介素 4 (IL-4) 的产生并不显着； (v) QFT-G 测定的灵敏度和特异性分别为 88.2% 和 18%，ICC 测定的灵敏度和特异性分别为 94.1% 和 36.4%。由于与非分枝杆菌的交叉反应，血液检测的特异性可能被低估。结核分枝杆菌感染，并且缺乏可用于诊断潜伏结核分枝杆菌感染的确认测试。流式细胞术可准确定位活动性结核病期间负责细胞因子产生的免疫效应细胞库。 ICC 检测是诊断活动性结核病的另一个有用工具。
A prospective cross-sectional blinded study of 28 patients (21 male and 7 female patients; mean age, 44 years) with suspected active tuberculosis (TB) attending a TB and chest clinic is described. Blood was taken for immune cell enumeration, a whole-blood enzyme-linked immunosorbent assay (ELISA) for the detection of gamma interferon (IFN-gamma) by the QuantiFERON-TB Gold (QFT-G) assay, and intracellular cytokine flow cytometry (ICC) analysis; and sputum was simultaneously taken for bacteriological culture for Mycobacterium tuberculosis. Twelve healthy subjects were included as controls. The performance characteristics of the QFT-G and ICC assays for the detection of active TB were compared. Among the patients with active TB, we found (i) normal to slightly elevated peripheral CD4(+) and CD8(+) T-cell counts but a significant reduction in the number of NK cells; (ii) CD4(+) T cells were the major cell type producing IFN-gamma, a type 1 cytokine; (iii) small percentages of CD8(+) T cells were also primed for IFN-gamma production; (iv) the production of interleukin-4 (IL-4), a type 2 cytokine, was not prominent; and (v) the sensitivity and the specificity of the QFT-G assay were 88.2% and 18%, respectively, and those of the ICC assay were 94.1% and 36.4%, respectively. The specificities of the blood tests were likely underestimated due to cross-reaction to a non-M. tuberculosis mycobacterial infection and the lack of a confirmatory test that could be used to diagnose latent M. tuberculosis infection. Flow cytometry accurately locates the pool of immunological effector cells responsible for cytokine production during active TB. The ICC assay is an additional useful tool for the diagnosis of active TB.