title: Purification and immunophenotypic characterization of human B cells with regulatory functions.
abstract: The analysis of human B cell populations of the blood relies on the expression of surface markers, mainly CD19, CD24, CD38, and CD27. According to these surface markers, three main B cell subsets can be identified in the blood: immature transitional B cells (CD19(+)CD24(high)CD38(high)), naïve B cells (CD19(+)CD24(int)CD38(int)) that have not encountered an antigen, and memory B cells (CD19(+)CD27(+)). To date, human B cells with regulatory functions have been essentially described within the CD24(high)CD38(high) transitional B cell subset. CD24(high)CD38(high) transitional B cells are able to produce interleukin 10 (IL-10) and to regulate in vitro Th1 and Th17 CD4(+) T cell activation. Here, we provide the methods to analyze and purify the CD24(high)CD38(high) transitional B cell subset for further in vitro experiments. We also provide a reliable method to detect B cell IL-10 production using intracellular cytokine staining.
authors.affiliation: INSERM U976, Paris, France.
authors.forename: Adèle
authors.initials: A
authors.lastname: de Masson
authors.forename: Hélène
authors.initials: H
authors.lastname: Le Buanec
authors.forename: Jean-David
authors.initials: JD
authors.lastname: Bouaziz
mesh_terms: D051997:ADP-ribosyl Cyclase 1 / Q000032:analysis* / Q000276:immunology; D060151:B-Lymphocytes, Regulatory / Q000166:cytology* / Q000276:immunology*; D051927:CD24 Antigen / Q000032:analysis* / Q000276:immunology; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D016753:Interleukin-10 / Q000032:analysis / Q000276:immunology
chemical_list: D051927:CD24 Antigen; D016753:Interleukin-10; D051997:ADP-ribosyl Cyclase 1