title: Novel Strategy for Phenotypic Characterization of Human B Lymphocytes from Precursors to Effector Cells by Flow Cytometry.
abstract: A precise identification and phenotypic characterization of human B-cell subsets is of crucial importance in both basic research and medicine. In the literature, flow cytometry studies for the phenotypic characterization of B-lymphocytes are mainly focused on the description of a particular cell stage, or of specific cell stages observed in a single type of sample. In the present work, we propose a backbone of 6 antibodies (CD38, CD27, CD10, CD19, CD5 and CD45) and an efficient gating strategy to identify, in a single analysis tube, a large number of B-cell subsets covering the whole B-cell differentiation from precursors to memory and plasma cells. Furthermore, by adding two antibodies in an 8-color combination, our approach allows the analysis of the modulation of any cell surface marker of interest along B-cell differentiation. We thus developed a panel of seven 8-colour antibody combinations to phenotypically characterize B-cell subpopulations in bone marrow, peripheral blood, lymph node and cord blood samples. Beyond qualitative information provided by biparametric representations, we also quantified antigen expression on each of the identified B-cell subsets and we proposed a series of informative curves showing the modulation of seventeen cell surface markers along B-cell differentiation. Our approach by flow cytometry provides an efficient tool to obtain quantitative data on B-cell surface markers expression with a relative easy-to-handle technique that can be applied in routine explorations.
authors.affiliation: Laboratoire d'Immunologie, Département d'Hématologie, Oncogénétique et Immunologie, Pôle de Biologie, Grenoble University Hospital, Grenoble, France.|BNI, TIMC-IMAG, UMR 5525 CNRS, Grenoble, France.|Université Grenoble-Alpes, Grenoble, France.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Clavarino
authors.affiliation: Laboratoire d'Immunologie, Département d'Hématologie, Oncogénétique et Immunologie, Pôle de Biologie, Grenoble University Hospital, Grenoble, France.
authors.forename: Noémie
authors.initials: N
authors.lastname: Delouche
authors.affiliation: Laboratoire d'Hématologie, Département d'Hématologie, Oncogénétique et Immunologie, Pôle de Biologie, Grenoble University Hospital, Grenoble, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Vettier
authors.affiliation: TheREx, TIMC-IMAG, UMR 5525 CNRS, Grenoble, France.|Etablissement Français du Sang Rhônes-Alpes Auvergne, La Tronche, France.|Université Grenoble-Alpes, Grenoble, France.
authors.forename: David
authors.initials: D
authors.lastname: Laurin
authors.affiliation: Laboratoire d'Immunologie, Département d'Hématologie, Oncogénétique et Immunologie, Pôle de Biologie, Grenoble University Hospital, Grenoble, France.
authors.forename: Martine
authors.initials: M
authors.lastname: Pernollet
authors.affiliation: Laboratoire d'Immunologie, Département d'Hématologie, Oncogénétique et Immunologie, Pôle de Biologie, Grenoble University Hospital, Grenoble, France.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Raskovalova
authors.affiliation: Laboratoire d'Immunologie, Département d'Hématologie, Oncogénétique et Immunologie, Pôle de Biologie, Grenoble University Hospital, Grenoble, France.|BNI, TIMC-IMAG, UMR 5525 CNRS, Grenoble, France.|Université Grenoble-Alpes, Grenoble, France.
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Cesbron
authors.affiliation: Laboratoire d'Immunologie, Département d'Hématologie, Oncogénétique et Immunologie, Pôle de Biologie, Grenoble University Hospital, Grenoble, France.|BNI, TIMC-IMAG, UMR 5525 CNRS, Grenoble, France.|Université Grenoble-Alpes, Grenoble, France.
authors.forename: Chantal
authors.initials: C
authors.lastname: Dumestre-Pérard
authors.affiliation: Laboratoire d'Immunologie, Département d'Hématologie, Oncogénétique et Immunologie, Pôle de Biologie, Grenoble University Hospital, Grenoble, France.|Université Grenoble-Alpes, Grenoble, France.|CNRS UMR 5309 and INSERM U1209, Institut Albert Bonniot, Grenoble, France.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Jacob