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Comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse
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 differi...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5756325/ https://www.ncbi.nlm.nih.gov/pubmed/29311759 http://dx.doi.org/10.1186/s12907-017-0068-6 |
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author | Stricker-Krongrad, Alain Shoemake, Catherine Zhong, Miao Liu, Jason Bouchard, Guy |
author_facet | Stricker-Krongrad, Alain Shoemake, Catherine Zhong, Miao Liu, Jason Bouchard, Guy |
author_sort | Stricker-Krongrad, Alain |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5756325 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-57563252018-01-08 Comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse Stricker-Krongrad, Alain Shoemake, Catherine Zhong, Miao Liu, Jason Bouchard, Guy BMC Clin Pathol Research Article 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. BioMed Central 2018-01-06 /pmc/articles/PMC5756325/ /pubmed/29311759 http://dx.doi.org/10.1186/s12907-017-0068-6 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Stricker-Krongrad, Alain Shoemake, Catherine Zhong, Miao Liu, Jason Bouchard, Guy Comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse |
title | Comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse |
title_full | Comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse |
title_fullStr | Comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse |
title_full_unstemmed | Comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse |
title_short | Comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse |
title_sort | comparison of a microsphere-based platform with a multiplex flow cytometric assay for determination of circulating cytokines in the mouse |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5756325/ https://www.ncbi.nlm.nih.gov/pubmed/29311759 http://dx.doi.org/10.1186/s12907-017-0068-6 |
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