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首页> 美国卫生研究院文献>Clinical and Diagnostic Laboratory Immunology >Measurement of Cytokine Secretion Intracellular Protein Expression and mRNA in Resting and Stimulated Peripheral Blood Mononuclear Cells
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Measurement of Cytokine Secretion Intracellular Protein Expression and mRNA in Resting and Stimulated Peripheral Blood Mononuclear Cells

机译：静息和刺激的外周血单个核细胞中细胞因子分泌细胞内蛋白表达和mRNA的测量

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摘要

Quantitation of cytokine production is a valuable adjunct to standard immunologic assays in defining several pathologic processes. Nevertheless, there is little agreement about which tissues should be assayed, which type of assay should be performed, and which stimulation protocol should be used. As these types of assays enter the clinical arena, there is need for standardization. There is also a need to maximize the amount of information which may be derived from a single sample. We compared secreted interleukin 4 (IL-4), IL-2, IL-6, tumor necrosis factor alpha (TNF-α), and gamma interferon proteins as measured by enzyme-linked immunosorbent assay with intracellular cytokine production (IL-2 and gamma interferon) as detected by flow cytometry and quantitative competitive PCR for IL-2, IL-4, TNF-α, and gamma interferon mRNA and cDNA. Results from unstimulated cells and cells stimulated with phorbol myristate acetate, phytohemagglutinin, and phorbol myristate acetate plus phytohemagglutin were compared. All three methodologies detected significant stimulation of cytokine production. The combination of phytohemagglutinin and phorbol myristate acetate was overall the most-potent stimulus.
机译：在确定几种病理过程中，细胞因子产生的定量是标准免疫学测定的有价值的辅助手段。然而，关于应检测哪些组织，应执行哪种类型的检测以及应使用哪种刺激方案的共识很少。随着这些类型的测定进入临床领域，需要进行标准化。还需要最大化可以从单个样本获得的信息量。我们比较了分泌型白介素4（IL-4），IL-2，IL-6，肿瘤坏死因子α（TNF-α）和γ干扰素蛋白，通过酶联免疫吸附测定与细胞内细胞因子产生（IL-2和通过流式细胞仪和定量竞争PCR检测IL-2，IL-4，TNF-α以及γ干扰素mRNA和cDNA进行检测。比较了未刺激的细胞和醋酸佛豆蔻肉豆蔻酸酯，植物血凝素以及醋酸佛豆蔻肉豆蔻酸酯+植物血凝素刺激的细胞的结果。三种方法均检测到细胞因子产生的明显刺激。植物血凝素和豆蔻酸佛波醇的组合总体上是最有效的刺激。

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