Simultaneous detection of 15 human cytokines in a single sample of stimulated peripheral blood mononuclear cells
Simultaneous detection of 15 human cytokines in a single sample of stimulated peripheral blood mononuclear cells
复制标题
DOI：
10.1128/cdli.10.1.133-139.2003
复制
发表时间：
2003-01-01
期刊：
影响因子：
--
通讯作者：
Rijkers, GT
中科院分区：
文献类型：
--
作者：
de Jager, W;te Velthuis, H;Rijkers, GT
免疫系统细胞分泌的细胞因子可以改变免疫细胞或其他细胞的行为和特性。在炎症部位，几组细胞因子与免疫细胞相互作用，它们的联合作用往往比单一成分的功能更重要。常规技术，如酶联免疫吸附试验，通常需要大量细胞来表征激活的淋巴细胞的完整细胞因子谱。Bio-Rad实验室的Bio-Plex系统结合了三明治免疫分析的原理和Luminex基于荧光珠的技术。我们建立了一种同时检测抗原和有丝分裂原刺激的人外周血单个核细胞培养上清液中不同细胞因子的多重细胞因子分析方法。15种人类细胞因子(IL-1α、IL-1β、IL-2、IL-4、IL-5、IL-6、IL-8、IL-10、IL-12p70、IL-13、IL-15、IL-17、IL-18、γ干扰素和肿瘤坏死因子α)与一组健康人、类风湿关节炎患者和青少年特发性关节炎患者进行了验证。将多重分析与常规的酶联免疫吸附分析技术与该供体小组进行比较，结果显示所有细胞因子的相关系数在0.75到0.99之间。批内变异小于10%，而批间变异介于10%至22%之间。事实证明，这一多重系统是细胞因子定量的有力工具。它将提供一个更完整的图景，说明健康人和慢性炎症性疾病患者激活的淋巴细胞细胞因子谱之间的差异。
Cytokines secreted by cells of the immune system can alter the behavior and properties of immune or other cells. At a site of inflammation, sets of cytokines interact with immune cells, and their combined effect is often more important than the function of one isolated component. Conventional techniques, such as enzyme-linked immunosorbent assays, generally require large quantities of cells to characterize a complete cytokine profile of activated lymphocytes. The Bio-Plex system from Bio-Rad Laboratories combines the principle of a sandwich immunoassay with the Luminex fluorescent-bead-based technology. We developed a multiplex cytokine assay to detect different cytokines simultaneously in culture supernatant of human peripheral blood mononuclear cells stimulated with antigen and with mitogen. Fifteen human cytokines (interieukin 1alpha [IL-1alpha], IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-18, gamma interferon, and tumor necrosis factor alpha) were validated with a panel of healthy individuals, rheumatoid arthritis patients, and juvenile idiopathic arthritis patients. Comparing the multiplex assay with a regular enzyme-linked immunosorbent assay technique with this donor panel resulted in correlation coefficients for all cytokines ranging from 0.75 to 0.99. Intra-assay variance proved to be less then 10%, whereas interassay variability ranged between 10 and 22%. This multiplex system proved to be a powerful tool in the quantitation of cytokines. It will provide a more complete picture in differences between activated lymphocyte cytokine profiles from healthy individuals and those from patients with chronic inflammatory diseases.