title: Analysis of Intracellular Ca(2+) Mobilization in Human NK Cell Subsets by Flow Cytometry.
abstract: In signaling cascades downstream of NK cell activating receptor engagement, Ca(2+) ions are pivotal second messengers for NK cell cytotoxicity as well as cytokine production. Upon cellular activation, intracellular mobilization of Ca(2+) ions initially involves depletion of endoplasmic reticulum stores, leading to subsequent Ca(2+) influx through specific plasma membrane Ca(2+) release activated Ca(2+) channels. Multiple probes and assays for detecting intracellular Ca(2+) concentrations have been developed. With the advance of multiparameter flow cytometry instrumentation, a thorough analysis of signaling in specific NK cell subsets is possible. Here, a flow cytometric method for dynamic measurements of intracellular Ca(2+) concentrations in human NK cells subsets is detailed and discussed. This assay can be further adapted for specific scientific and diagnostic questions, with implications for various immunopathological conditions.
authors.affiliation: Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186, Stockholm, Sweden.
authors.forename: Jakob
authors.initials: J
authors.lastname: Theorell
authors.affiliation: Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 14186, Stockholm, Sweden. yenan.bryceson@ki.se.|Broegelmann Research Laboratory, Department of Clinical Science, University of Bergen, 5021, Bergen, Norway. yenan.bryceson@ki.se.
authors.forename: Yenan T
authors.initials: YT
authors.lastname: Bryceson
mesh_terms: D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D008213:Lymphocyte Activation; D015398:Signal Transduction
chemical_list: D015220:Calcium Channels; D002118:Calcium
keywords: Activation; Ca2+ flux; Human; Natural killer cells; Polychromatic flow cytometry