Comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse.

Q2 Medicine
BMC Clinical Pathology Pub Date : 2018-01-06 eCollection Date: 2018-01-01 DOI:10.1186/s12907-017-0068-6
Alain Stricker-Krongrad, Catherine Shoemake, Miao Zhong, Jason Liu, Guy Bouchard
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引用次数: 0

Abstract

Background: Measuring expression profiles of inflammatory biomarkers is important in monitoring the polarization of immune responses; therefore, results should be independent of quantitation methods if they are to be accepted as validated clinical pathology biomarkers. To evaluate effects of differing quantitation methods, the seven major circulating Th1/Th2/Th17 cytokines interleukin 2 (IL-2), interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), IL-4, IL-6, IL-10 and IL-17A were quantified in plasma of lipopolysaccharide (LPS)-treated mice with two different multiplex platforms.

Methods: Female C57BL6 mice were treated orally with vehicle or dexamethasone, followed by LPS intravenously. Plasma samples were analyzed 0.5, 1, 2, 4, and 6 h post-LPS challenge with assays at Myriad-RBM and compared to assays performed on a BD Accuri C6 flow cytometer.

Results: IL-17A response to LPS was limited but sustained, and the response for the remaining cytokines were early and transient; dexamethasone reduced expression of all 7 cytokines. TNF-α and IL-6 levels were similar across both assays, and IL-4 levels were generally very low. Plasma levels of remaining cytokines were variably lower with BD assays than Myriad-RBM assays.

Conclusions: The present findings demonstrate that quantitation of circulating biomarkers of inflammation can be achieved using multiplexed flow cytometry, but careful consideration must be taken for assay validation when cross-referencing with another multiplexed assay.

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微球平台与多重流式细胞术测定小鼠循环细胞因子的比较。
背景:测量炎症生物标志物的表达谱对于监测免疫反应的极化是重要的;因此,如果结果要被接受为有效的临床病理生物标志物,那么结果应该独立于定量方法。为了评价不同定量方法的效果,我们采用两种不同的复合平台对脂多糖(LPS)处理小鼠血浆中7种主要的循环Th1/Th2/Th17细胞因子白介素2 (IL-2)、干扰素γ (IFN-γ)、肿瘤坏死因子α (TNF-α)、IL-4、IL-6、IL-10和IL-17A进行了定量。方法:雌性C57BL6小鼠口服给药或地塞米松,然后静脉注射LPS。血浆样品在lps刺激后0.5、1、2、4和6小时用Myriad-RBM进行分析,并与在BD Accuri C6流式细胞仪上进行的分析进行比较。结果:IL-17A对LPS的反应有限但持续,对其余细胞因子的反应早期且短暂;地塞米松降低了7种细胞因子的表达。两种检测中TNF-α和IL-6水平相似,IL-4水平普遍很低。与Myriad-RBM相比,BD检测的血浆剩余细胞因子水平有所降低。结论:目前的研究结果表明,使用多路流式细胞术可以实现炎症循环生物标志物的定量,但在与另一种多路流式细胞术交叉对照时,必须仔细考虑分析验证。
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来源期刊
BMC Clinical Pathology
BMC Clinical Pathology Medicine-Pathology and Forensic Medicine
CiteScore
3.30
自引率
0.00%
发文量
0
期刊介绍: BMC Clinical Pathology is an open access journal publishing original peer-reviewed research articles in all aspects of histopathology, haematology, clinical biochemistry, and medical microbiology (including virology, parasitology, and infection control). BMC Clinical Pathology (ISSN 1472-6890) is indexed/tracked/covered by PubMed, CAS, EMBASE, Scopus and Google Scholar.
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