title: Optimized flow cytometry protocol for analysis of surface expression of interleukin-1 receptor types I and II.
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.
authors: 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, vasilyevmd@gmail.com.
authors: Filipp Filippovich
authors: FF
authors: Vasilyev
authors: Julia Anatolievna
authors: JA
authors: Lopatnikova
authors: Sergey Vitalievich
authors: SV
authors: Sennikov