A comparison of biofluid cytokine markers across platform technologies: Correspondence or divergence?
A comparison of biofluid cytokine markers across platform technologies: Correspondence or divergence?
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DOI：
10.1016/j.cyto.2018.05.032
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发表时间：
2018-11-01
期刊：
影响因子：
3.8
通讯作者：
Kramer, J. H.
中科院分区：
文献类型：
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作者：
Casaletto, K. B.;Elahi, F. M.;Kramer, J. H.
背景：生物流体细胞因子的定量是转化研究的一个快速发展的领域。然而，用于检测相同蛋白质的越来越多的可用检测平台之间的可比性仍有待确定。我们的目的是通过两个高灵敏度定量平台直接比较典型老年人血浆中一组常用的细胞因子，即 Meso Scale Discovery 高性能电化学发光 (HPE) 和Quanterix 的单分子免疫吸附测定 (Simoa)。方法：57 名社区居住的老年人完成了抽血、神经心理学评估和脑 MRI，作为健康大脑衰老研究的一部分。在 HPE 和 Simoa 上分别对同一抽取日期的血浆样本进行 IL-10、IP-10、IL-6、TNF α 和 IL-1 β 分析。检查了可靠的可检测性（方差系数 (CV) < 20%，异常值高于中位数 3 个四分位数）、测定内精度、绝对浓度、跨平台的可重复性以及与感兴趣的外部变量（例如人口统计、血管健康的外周标志物和大脑健康）的并发关联。 结果：在 HPE (87.7-93.0%) 和 Simoa 上可靠测量的细胞因子比例(75.4-93.0%) 没有差异 (ps > 0.32)，但 IL-1 beta 除外，它只能使用 Simoa (68.4%) 进行可靠测量。平均而言，两个平台的 CV 都在 < 8% 的范围内，是可以接受的。使用 Simoa 测得的 IL-10、IL-6 和 TNF α 的绝对浓度较高 (ps < 0.05)。 HPE 和 Simoa 在相同细胞因子蛋白上彼此之间仅存在小到中等比例的差异（范围：IL-10 的 r = 0.26 到 IL-6 的 r = 0.64），但平台一致性并不依赖于细胞因子浓度。每个平台内的细胞因子比率在 HPE 和 Simoa 中表现出相似的上调和下调相对模式，但仍存在显着差异 (ps < 0.001)。支持并发有效性，细胞因子和外部变量之间相关性的所有 95% 置信区间在两个平台之间重叠。此外，大多数关联都符合预期方向，并且跨平台一致（例如，IL-6 和 TNF α），但 IP-10 和 IL-10 有几个明显的例外。结论：HPE 和 Simoa 在测量一组常用细胞因子蛋白时表现出可比的可检测性和测定内精度，但 IL-1 β 除外，IL-1 β 在 HPE 上无法可靠检测。然而，Simoa 表现出整体较高的浓度，并且两个平台在直接比较时并未表现出一致性。相对细胞因子比率和关联在不同平台上表现出相似的模式。绝对细胞因子浓度可能无法在平台之间直接比较，可能取决于分析物，并且解释可能最好限于相对关联的讨论。
Background: Quantification of biofluid cytokines is a rapidly growing area of translational research. However, comparability across the expanding number of available assay platforms for detection of the same proteins remains to be determined. We aimed to directly compare a panel of commonly measured cytokines in plasma of typically aging adults across two high sensitivity quantification platforms, Meso Scale Discovery high performance electrochemiluminiscence (HPE) and single-molecule immunosorbent assays (Simoa) by Quanterix.Methods: 57 community-dwelling older adults completed a blood draw, neuropsychological assessment, and brain MRI as part of a healthy brain aging study. Plasma samples from the same draw dates were analyzed for IL-10, IP-10, IL-6, TNF alpha, and IL-1 beta on HPE and Simoa, separately. Reliable detectability (coefficient of variance (CV) < 20% and outliers 3 interquartiles above the median removed), intra-assay precision, absolute concentrations, reproducibility across platforms, and concurrent associations with external variables of interest (e.g., demographics, peripheral markers of vascular health, and brain health) were examined.Results: The proportion of cytokines reliably measured on HPE (87.7-93.0%) and Simoa (75.4-93.0%) did not differ (ps > 0.32), with the exception of IL-1 beta which was only reliably measured using Simoa (68.4%). On average, CVs were acceptable at < 8% across both platforms. Absolute measured concentrations were higher using Simoa for IL-10, IL-6, and TNF alpha (ps < 0.05). HPE and Simoa shared only small-to-moderate proportions of variance with one another on the same cytokine proteins (range: r = 0.26 for IL-10 to r = 0.64 for IL-6), though platform agreement did not dependent on cytokine concentrations. Cytokine ratios within each platform demonstrated similar relative patterns of up- and down-regulation across HPE and Simoa, though still significantly differed (ps < 0.001). Supporting concurrent validity, all 95% confidence intervals of the correlations between cytokines and external variables overlapped between the two platforms. Moreover, most associations were in expected directions and consistently so across platforms (e.g., IL-6 and TNF alpha), though with several notable exceptions for IP-10 and IL-10.Conclusions: HPE and Simoa showed comparable detectability and intra-assay precision measuring a panel of commonly examined cytokine proteins, with the exception of IL-1 beta which was not reliably detected on HPE. However, Simoa demonstrated overall higher concentrations and the two platforms did not show agreement when directly compared against one another. Relative cytokine ratios and associations demonstrated similar patterns across platforms. Absolute cytokine concentrations may not be directly comparable across platforms, may be analyte dependent, and interpretation may be best limited to discussion of relative associations.