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

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

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作者:Alain Stricker-Krongrad, Catherine Shoemake, Miao Zhong, Jason Liu, Guy Bouchard

Background

Measuring expression profiles of inflammatory biomarkers is important in monitoring the polarization of immune responses; therefore,

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.

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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