title: Multi-center harmonization of flow cytometers in the context of the European "PRECISESADS" project.
abstract: The innovative medicine initiative project called PRECISESADS will study 2.500 individuals affected by systemic autoimmune diseases (SADs) and controls. Among extensive OMICS approaches, multi-parameter flow cytometry analyses will be performed in eleven different centers. Therefore, the integration of all data in common bioinformatical and biostatistical investigations requires a fine mirroring of all instruments. We describe here the procedure elaborated to achieve this prerequisite. One flow cytometer chosen as reference instrument fixed the mean fluorescence intensities (MFIs) of 8 different fluorochrome-conjugated antibodies (Abs) using VersaComp Ab capture beads. The ten other centers adjusted their own PMT voltages to reach the same MFIs. Subsequently, all centers acquired Rainbow 8-peak beads data on a daily basis to follow the stability of their instrument overtime. One blood sample has been dispatched and concomitantly stained in all centers. Comparison of leukocytes frequencies and cell surface marker MFIs demonstrated the close sensitivity of all flow cytometers, allowing a multicenter analysis. The effective multi-center harmonization enables the constitution of a workable wide flow cytometry database for the identification of specific molecular signatures in individuals with SADs.
authors.affiliation: INSERM ERI29, EA2216, Université de Brest, Labex IGO, CHRU Morvan, Brest, France. Electronic address: christophe.jamin@univ-brest.fr.
authors.forename: Christophe
authors.initials: C
authors.lastname: Jamin
authors.affiliation: INSERM ERI29, EA2216, Université de Brest, Labex IGO, CHRU Morvan, Brest, France.
authors.forename: Lucas
authors.initials: L
authors.lastname: Le Lann
authors.affiliation: Chromatin and Disease Group, Cancer Epigenetics and Biology Programme (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), L'Hospitalet de Llobregat Barcelona, Spain.
authors.forename: Damiana
authors.initials: D
authors.lastname: Alvarez-Errico
authors.affiliation: IMIBIC/Reina Sofia Hospital/University of Cordoba, Córdoba, Spain.
authors.forename: Nuria
authors.initials: N
authors.lastname: Barbarroja
authors.affiliation: Laboratory of Tissue Homeostasis and Disease, Faculty of Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Tineke
authors.initials: T
authors.lastname: Cantaert
authors.affiliation: Pôle de Pathologies Rhumatismales, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Julie
authors.initials: J
authors.lastname: Ducreux
authors.affiliation: Department of Immunology and Allergy, University Hospital and School of Medicine, Geneva, Switzerland; Department of Pathology and Immunology, School of Medicine, Geneva, Switzerland.
authors.forename: Aleksandra Maria
authors.initials: AM
authors.lastname: Dufour
authors.affiliation: Department of Rheumatology and Clinical Immunology, Charité Hospital, Berlin, Germany.
authors.forename: Velia
authors.initials: V
authors.lastname: Gerl
authors.affiliation: Klinik für Immunologie und Rheumatologie, Medizinische Hochschule Hannover, Hannover, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Kniesch
authors.affiliation: Serviço de Imunologia EX-CICAP, Centro Hospitalar do Porto, Porto, Portugal.
authors.forename: Esmeralda
authors.initials: E
authors.lastname: Neves
authors.affiliation: Laboratorio di Analisi Chimico Cliniche e Microbiologia-Servizio di Citofluorimetria, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico di Milano, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Trombetta
authors.affiliation: GENYO, Centre for Genomics and Oncological Research Pfizer, University of Granda, Andalusian Regional Government, PTS GRANADA, Granada, Spain; Institute for Environmental Medicine, Karolinska Institutet Stockholm, Sweeden.
authors.forename: Marta
authors.initials: M
authors.lastname: Alarcón-Riquelme
authors.affiliation: GENYO, Centre for Genomics and Oncological Research Pfizer, University of Granda, Andalusian Regional Government, PTS GRANADA, Granada, Spain.
authors.forename: Concepción
authors.initials: C
authors.lastname: Marañon
authors.affiliation: INSERM ERI29, EA2216, Université de Brest, Labex IGO, CHRU Morvan, Brest, France.
authors.forename: Jacques-Olivier
authors.initials: JO
authors.lastname: Pers
mesh_terms: D000906:Antibodies; D001327:Autoimmune Diseases / Q000097:blood / Q000145:classification*; D002138:Calibration; D005060:Europe; D005434:Flow Cytometry / Q000295:instrumentation* / Q000592:standards; D006801:Humans
chemical_list: D000906:Antibodies
keywords: Multi-center harmonization; Multi-parametric flow cytometry; PRECISESADS; Standard operating procedure