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      Multi-center harmonization of flow cytometers in the context of the European "PRECISESADS" project.

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          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.

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          Author and article information

          Journal
          Autoimmun Rev
          Autoimmunity reviews
          Elsevier BV
          1873-0183
          1568-9972
          Nov 2016
          : 15
          : 11
          Affiliations
          [1 ] INSERM ERI29, EA2216, Université de Brest, Labex IGO, CHRU Morvan, Brest, France. Electronic address: christophe.jamin@univ-brest.fr.
          [2 ] INSERM ERI29, EA2216, Université de Brest, Labex IGO, CHRU Morvan, Brest, France.
          [3 ] Chromatin and Disease Group, Cancer Epigenetics and Biology Programme (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), L'Hospitalet de Llobregat Barcelona, Spain.
          [4 ] IMIBIC/Reina Sofia Hospital/University of Cordoba, Córdoba, Spain.
          [5 ] Laboratory of Tissue Homeostasis and Disease, Faculty of Medicine, KU Leuven, Leuven, Belgium.
          [6 ] Pôle de Pathologies Rhumatismales, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
          [7 ] Department of Immunology and Allergy, University Hospital and School of Medicine, Geneva, Switzerland; Department of Pathology and Immunology, School of Medicine, Geneva, Switzerland.
          [8 ] Department of Rheumatology and Clinical Immunology, Charité Hospital, Berlin, Germany.
          [9 ] Klinik für Immunologie und Rheumatologie, Medizinische Hochschule Hannover, Hannover, Germany.
          [10 ] Serviço de Imunologia EX-CICAP, Centro Hospitalar do Porto, Porto, Portugal.
          [11 ] Laboratorio di Analisi Chimico Cliniche e Microbiologia-Servizio di Citofluorimetria, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico di Milano, Italy.
          [12 ] 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.
          [13 ] GENYO, Centre for Genomics and Oncological Research Pfizer, University of Granda, Andalusian Regional Government, PTS GRANADA, Granada, Spain.
          Article
          S1568-9972(16)30179-3
          10.1016/j.autrev.2016.07.034
          27490203
          630d5d8c-b709-4692-aafa-be09d9259756
          History

          Multi-center harmonization,Multi-parametric flow cytometry,PRECISESADS,Standard operating procedure

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