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首页> 外文期刊>Cytokine >Effect of anticoagulants on multiplexed measurement of cytokine/chemokines in healthy subjects
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Effect of anticoagulants on multiplexed measurement of cytokine/chemokines in healthy subjects

机译：抗凝剂对健康受试者细胞因子/趋化因子的多重测量的影响

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Introduction: Cytokines are humoral regulatory molecules that act together in immunologic pathways. Monitoring cytokines and their variations within physiologic ranges is critical for biomarker discovery. Therefore, we evaluated the performance characteristics of 72 analytes measured by multiplex cytokine immunoassay, with an emphasis on the differences of analytes measured in serum compared to plasma, and, in plasma, on the impact of anticoagulants on the cytokine measurement. Methods: We used fluorescent bead-based (Luminex) immunoassay kits to simultaneously measure 72 analytes. We tested serum and plasma samples from 11 matched donors. Plasma samples were anti-coagulated with sodium heparin, sodium citrate dextrose and ethylene diamine tetra-acetic acid (EDTA), respectively. Results: Of the 72 cytokines, 12 were undetectable in all types of specimen samples. Nineteen analytes, including PDGF-bb, IL-4, IL-8, IL-9, FGF-b, PAI-1, CXCL-5, CCL-5, CD40L, EGF, VEGF, IL-2ra, IL-3, SDF-1a, PCT, MCP-3, GIP, IL-16 and fibrinogen, showed significant differences between measurements in serum and all types of plasma, regardless of anticoagulant. Among plasma samples, 10 analytes (eotaxin, SCGF-b, MCP-1, SCF, MIP-1b, VEGF, RANTES, PDGF-b, PAI-1 and ITAC) showed significantly higher concentrations in heparinized plasma compared to citrated and EDTA plasma. IP-10, and CTAK were the only 2 cytokines that presented different concentrations in citrate and EDTA plasma. Conclusions: With their small volume, low cost per test, and multiplex capacity, Luminex-based cytokine assays have enormous potential utility for screening in epidemiologic studies. In our study, we showed that many cytokines' concentrations differed between serum and plasma samples, and that different anticoagulants used in preparation of plasma samples also affected the measurement of some cytokines. There was no optimal sample preparation that was clearly superior for the measurement of all analytes measured. Ultimately, the utility of cytokine measurement, as biomarker or to monitor the immune system, will depend on attention to detail in the collection and processing of samples in addition to assay precision.
机译：简介：细胞因子是在免疫途径中共同起作用的体液调节分子。监测细胞因子及其在生理范围内的变化对于发现生物标志物至关重要。因此，我们评估了通过多重细胞因子免疫测定法测量的72种分析物的性能特征，重点是与血浆相比，在血清中测量的分析物的差异，以及在血浆中，抗凝剂对细胞因子测量的影响。方法：我们使用基于荧光珠的（Luminex）免疫测定试剂盒来同时测量72种分析物。我们测试了来自11个匹配供体的血清和血浆样品。血浆样品分别用肝素钠，柠檬酸钠和右旋乙二胺四乙酸（EDTA）进行抗凝。结果：在72种细胞因子中，在所有类型的样本样品中均未检测到12种。 19种分析物，包括PDGF-bb，IL-4，IL-8，IL-9，FGF-b，PAI-1，CXCL-5，CCL-5，CD40L，EGF，VEGF，IL-2ra，IL-3， SDF-1a，PCT，MCP-3，GIP，IL-16和纤维蛋白原显示血清和所有类型血浆的测量值之间存在显着差异，而与抗凝剂无关。在血浆样品中，与柠檬酸盐和EDTA血浆相比，肝素化血浆中的10种分析物（eotaxin，SCGF-b，MCP-1，SCF，MIP-1b，VEGF，RANTES，PDGF-b，PAI-1和ITAC）的浓度明显更高。 IP-10和CTAK是仅有的2种在柠檬酸盐和EDTA血浆中呈现不同浓度的细胞因子。结论：基于Luminex的细胞因子测定法体积小，每次测试成本低且具有多重能力，在流行病学研究中具有巨大的筛选潜力。在我们的研究中，我们显示了血清和血浆样品之间许多细胞因子的浓度不同，并且血浆样品制备中使用的不同抗凝剂也影响了某些细胞因子的测定。没有最佳的样品前处理明显优于所有被测分析物。最终，细胞因子测量作为生物标志物或监测免疫系统的效用将取决于对样品收集和处理过程中细节的关注，以及测定的准确性。

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