| Correlation of XMAP and ELISA cytokine profiles; development and validation for immunotoxicological studies in vitro. | |
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MedLine Citation:
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PMID: 21053779 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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There is an emerging trend in immunotoxicological studies to use the multiplex technologies for testing the safety and the efficacy of new pharmaceuticals by using cytokines profiling as biomarker. The Luminex 200 xMAP (multi-analyte profiling) technology provides simultaneous measurement of multiple cytokines in small sample volumes, expressing rapidly the differences between various test compounds. The aim is to develop and validate the Luminex 200 multiplex immunoassays by correlation with ELISA (enzyme-linked immunosorbent assays) for implementation in evaluating cytokine profiling in immunotoxicological studies in vitro. METHODS: Human peripheral whole blood from healthy subject diluted 1+4 with RPMI 1640 was cultured 48 hours in 28 experimental variants: control, in presence of mitogens, bioflavonoid extracts (from Crataegus monogyna and Echinacea purpurea) as cytoprotectors and with a toxic compound [Pb(NO3)2]), separately or variously combined. IL-1beta and IL-2 were comparatively performed by xMAP and ELISA immunoassays from the same sample to initialize validation of multiplex cytokine panel: IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-alpha, IFN-gamma, usually performed by Luminex 200 system in our immunotoxicological studies. The results indicate similarly typed trends of cytokine values obtained by both methods, with comparable relative changes in presence of mitogens, bioflavonoids and toxic, respectively. Although xMAP absolute cytokine values were higher than ELISA values, the correlation between multiplexed assay and ELISA was good for IL-1beta and IL-2 with positive correlation coefficients near to 1. Conclusions. Quantitative differences between absolute values for IL-1beta and IL-2 obtained by xMAP and ELISA assays are found, but the relative values are comparable and the two methods keep similar trends in similar exposure conditions. The performance parameters of the xMAP assay and the good correlation coefficients with the "gold standard" ELISA recommend to validate the multiplex assay for analyzing cytokine profiles in immunotoxicological studies in vitro. |
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Authors:
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Eleonora Codorean; Cornelia Nichita; Lucian Albulescu; Elena Răducan; Ionela Daniela Popescu; Alina Constantina Lonită; Radu Albulescu |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Validation Studies |
Journal Detail:
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Title: Roumanian archives of microbiology and immunology Volume: 69 ISSN: 1222-3891 ISO Abbreviation: Roum Arch Microbiol Immunol Publication Date: 2010 Jan-Mar |
Date Detail:
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Created Date: 2010-11-08 Completed Date: 2010-12-14 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9204717 Medline TA: Roum Arch Microbiol Immunol Country: Romania |
Other Details:
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Languages: eng Pagination: 13-9 Citation Subset: IM |
Affiliation:
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Victor Babes National Institute of Pathology, Bucharest. imunotox@vbabes.ro |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Biological Markers
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analysis Cells, Cultured Cytokines / analysis* Enzyme-Linked Immunosorbent Assay Flavonoids / immunology Humans Immunoassay / methods*, standards* Interleukin-1beta / analysis Interleukin-2 / analysis Leukocytes, Mononuclear Reagent Kits, Diagnostic / standards* Sensitivity and Specificity |
| Chemical | |
Reg. No./Substance:
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0/Biological Markers; 0/Cytokines; 0/Flavonoids; 0/Interleukin-1beta; 0/Interleukin-2; 0/Reagent Kits, Diagnostic |
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