Analysis of inflammatory cytokines in human blood, breath condensate, and urine using a multiplex immunoassay platform
Analysis of inflammatory cytokines in human blood, breath condensate, and urine using a multiplex immunoassay platform
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DOI：
10.3109/1354750x.2014.988646
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发表时间：
2015-02-01
期刊：
影响因子：
2.6
通讯作者：
Madden, Michael C.
中科院分区：
文献类型：
--
作者：
Stiegel, Matthew A.;Pleil, Joachim D.;Madden, Michael C.
人类生物介质中细胞因子表达的变化表明对外部应激源的炎症反应，并反映了各种健康终点的不良结局途径（AOP）的早期沿着步骤。为了表征和解释这种炎症反应，开发了使用电致发光多重Th 1/Th 2细胞因子免疫测定平台测量人血液、呼出气冷凝物（EBC）和尿液中的一套10种不同细胞因子的方法。使用2008-2012年从北卡罗来纳州名义上健康的成年受试者中采集的90份血浆、77份EBC和400份尿液样本，评价了8种白细胞介素（IL）（1b、2、4、5、8、10、12 p70和13）、干扰素-γ（IFN-γ）和肿瘤坏死因子-α（TNF-α）的测量分布和相关性。体内结果表明，在所有三种生物培养基中，在0.05-0.10 rho g/ml的水平下具有足够的灵敏度来表征所有10种细胞因子，动态范围高达100 ng/ml。体内测量结果还表明，血液、EBC和尿液样本的重复分析的平均估计倍数范围分别为2.21、3.49和2.50，与平均估计倍数范围相似（2.88）最低浓度（0.610 rho g/ml）;细胞因子方法可用于所有三种生物培养基。在EBC中测量的10种细胞因子中有9种彼此高度相关，斯皮尔曼rho系数范围为0.679至0.852，而在血液中测量的细胞因子具有负相关和正相关的混合，范围为-0.620至0.836。EBC与血液几乎全部呈正相关。这项工作也代表了血液，EBC和尿液中超痕量炎症标志物的首次成功的人内和人与人之间的评估。
A change in the expression of cytokines in human biological media indicates an inflammatory response to external stressors and reflects an early step along the adverse outcome pathway (AOP) for various health endpoints. To characterize and interpret this inflammatory response, methodology was developed for measuring a suite of 10 different cytokines in human blood, exhaled breath condensate (EBC), and urine using an electrochemiluminescent multiplex Th1/Th2 cytokine immunoassay platform. Measurement distributions and correlations for eight interleukins (IL) (1b, 2, 4, 5, 8, 10, 12p70 and 13), interferon-gamma (IFN-gamma), and tumor necrosis factor-a (TNF-alpha) were evaluated using 90 blood plasma, 77 EBC, and 400 urine samples collected from nominally healthy adults subjects in North Carolina in 2008-2012. The in vivo results show that there is sufficient sensitivity for characterizing all 10 cytokines at levels of 0.05-0.10 rho g/ml with a dynamic range up to 100 ng/ml across all three of these biological media. The measured in vivo results also show that the duplicate analysis of blood, EBC and urine samples have average estimated fold ranges of 2.21, 3.49, and 2.50, respectively, which are similar to the mean estimated fold range (2.88) for the lowest concentration (0.610 rho g/ml) from a series of spiked control samples; the cytokine method can be used for all three biological media. Nine out of the 10 cytokines measured in EBC were highly correlated within one another with Spearman rho coefficients ranging from 0.679 to 0.852, while the cytokines measured in blood had a mix of negative and positive correlations, ranging from -0.620 to 0.836. Almost all correlations between EBC and blood were positive. This work also represents the first successful within-and between-person evaluation of ultra trace-level inflammatory markers in blood, EBC, and urine.