Optimized flow cytometry protocol for analysis of surface expression of interleukin-1 receptor types I and II |
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Authors: | Filipp Filippovich Vasilyev Julia Anatolievna Lopatnikova Sergey Vitalievich Sennikov |
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Affiliation: | 1. Laboratory of Molecular Immunology, Federal State Budgetary Institution “Research Institute of Clinical Immunology”, Russian Academy of Medical Sciences Siberian Branch, 14, Yadrincevskaja str., 630099, Novosibirsk, Russia
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Abstract: | The biological effects of interleukin (IL)-1 are realized through binding to specific membrane-bound receptors. The efficiency of IL-1 action depends on the number of receptors on the cell. We determined the percentage of cells that express IL-1 receptor type I (IL-1RI) and IL-1 receptor type II (IL-1RII) by flow cytometry using phycoerythrin (PE)-labelled antibodies to the IL-1Rs, and the mean absolute number of membrane-bound IL-1Rs per cell using QuantiBRITE PE calibration beads. We showed that different subpopulations of immunocompetent cells expressed different numbers of molecules of membrane-bound IL-1RI and IL-1RII. We also established that when cells were stimulated with bacterial lipopolysaccharide, there was a significant increase in the number of IL-1RI expressed, and a significant decrease in the mean number of IL-1RII molecules per cell. Determination of the mean number of membrane-bound IL-1R molecules using this protocol enables us to obtain precise and reproducible data that are necessary for full evaluation of expression levels. |
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Keywords: | Flow cytometry Interleukin-1 receptor Quantitative analysis Peripheral blood mononuclear cells |
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