title: Purification of Human Dendritic Cell Subsets from Peripheral Blood.
abstract: Blood represents the most accessible source of human dendritic cells (DCs). We present here a method to isolate three DC subtypes, as identified until now, from peripheral blood: plasmacytoid dendritic cells (pDCs), CD141(+) myeloid DCs, and CD1c(+) myeloid DCs. The method is based on the sequential depletion of non-DCs. First, depletion of granulocytes, erythrocytes, and platelets is obtained by blood centrifugation over a Ficoll gradient. Then, antibodies recognizing non-DCs, combined with magnetic beads, allow enrichment of DCs from peripheral blood mononuclear cells (PBMCs). Finally, enriched DCs are purified and separated into the different subtypes by immunolabeling and fluorescence-activated cell sorting (FACS) using DC-specific surface markers.DC studies might contribute to the comprehension of human immune processes in physiological and pathological conditions. Human blood DCs targeting might be a useful tool to ameliorate inflammatory diseases and improve vaccination strategies.
authors.affiliation: INSERM U932 Immunity and Cancer, 26 Rue D'Ulm, 75005, Paris, France.|Institut Curie, Centre de recherche, 26 Rue D'Ulm, 75005, Paris, France.
authors.forename: Solana
authors.initials: S
authors.lastname: Alculumbre
authors.affiliation: INSERM U932 Immunity and Cancer, 26 Rue D'Ulm, 75005, Paris, France. Lucia.pattarini@curie.fr.|Institut Curie, Centre de recherche, 26 Rue D'Ulm, 75005, Paris, France. Lucia.pattarini@curie.fr.
authors.forename: Lucia
authors.initials: L
authors.lastname: Pattarini
mesh_terms: D018949:Antigens, CD1 / Q000378:metabolism; D000954:Antigens, Surface / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology; D005434:Flow Cytometry; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000276:immunology; D018180:Thrombomodulin
chemical_list: D018949:Antigens, CD1; D000954:Antigens, Surface; C437944:CD1C protein, human; D006023:Glycoproteins; C546931:THBD protein, human; D018180:Thrombomodulin
keywords: Blood; Dendritic cell; Human; Subtypes; pDC