title: Identification of autoreactive B cells with labeled nucleosomes.
abstract: The pathogenesis of autoimmune diseases has not been completely elucidated yet, and only a few specific treatments have been developed so far. In autoimmune diseases mediated by pathogenic autoantibodies, such as systemic lupus erythematosus, the specific detection and analysis of autoreactive B cells is crucial for a better understanding of the physiopathology. Biological characterization of these cells may help to define new therapeutic targets. Very few techniques allowing the precise detection of autoreactive B cells have been described so far. Herein we propose a new flow cytometry technique for specific detection of anti-nucleosome B cells, which secrete autoantibodies in systemic lupus erythematosus, using labeled nucleosomes. We produced different fluorochrome-labeled nucleosomes, characterized them, and finally tested them in flow cytometry. Nucleosomes labeled via the cysteines present in H3 histone specifically bind to autoreactive B cells in the anti-DNA transgenic B6.56R mice model. The present work validates the use of fluorochrome-labeled nucleosomes via cysteines to identify anti-nucleosome B cells and offers new opportunities for the description of autoreactive B cell phenotype.
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France.|Department of Clinical Immunology and Internal Medicine, National Reference Center for Autoimmune Diseases, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Gies
authors.affiliation: Laboratory of Functional ChemoSystems, CNRS-University of Strasbourg UMR 7199/Laboratory of Excellence MEDALIS, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67400, Illkirch, France.
authors.forename: Alain
authors.identifier: http://orcid.org/0000-0003-3125-601X
authors.initials: A
authors.lastname: Wagner
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Seifert
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France.|Department of Clinical Immunology and Internal Medicine, National Reference Center for Autoimmune Diseases, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Guffroy
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France.
authors.forename: Jean-D
authors.initials: JD
authors.lastname: Fauny
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France.
authors.forename: Anne-M
authors.initials: AM
authors.lastname: Knapp
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France.|Department of Clinical Immunology and Internal Medicine, National Reference Center for Autoimmune Diseases, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.|UFR Médecine, Université de Strasbourg, Strasbourg, France.
authors.forename: Jean-L
authors.initials: JL
authors.lastname: Pasquali
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France.|Department of Clinical Immunology and Internal Medicine, National Reference Center for Autoimmune Diseases, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.|UFR Médecine, Université de Strasbourg, Strasbourg, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Martin
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Dumortier
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France.|Department of Clinical Immunology and Internal Medicine, National Reference Center for Autoimmune Diseases, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.|UFR Médecine, Université de Strasbourg, Strasbourg, France.
authors.forename: Anne-S
authors.initials: AS
authors.lastname: Korganow
authors.affiliation: CNRS UPR 3572 "Immunopathology and Therapeutic Chemistry"/Laboratory of Excellence Medalis, Institute of Molecular and Cellular Biology (IBMC), Strasbourg, France. pauline.soulas@ibmc-cnrs.unistra.fr.|Department of Clinical Immunology and Internal Medicine, National Reference Center for Autoimmune Diseases, Hôpitaux Universitaires de Strasbourg, Strasbourg, France. pauline.soulas@ibmc-cnrs.unistra.fr.|UFR Sciences pharmaceutiques, Université de Strasbourg, Illkirch-Graffenstaden, France. pauline.soulas@ibmc-cnrs.unistra.fr.
authors.forename: Pauline
authors.initials: P
authors.lastname: Soulas-Sprauel
mesh_terms: D000818:Animals; D001323:Autoantibodies / Q000276:immunology; D001324:Autoantigens / Q000276:immunology; D015551:Autoimmunity; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D015415:Biomarkers; D002460:Cell Line; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000276:immunology / Q000378:metabolism; D051379:Mice; D009707:Nucleosomes / Q000276:immunology* / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D015415:Biomarkers; D009707:Nucleosomes