1. Semi-automated and standardized cytometric procedures for multi-panel and multi-parametric whole blood immunophenotyping
- Author
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Hasan, Milena, Beitz, Benoit, Rouilly, Vincent, Libri, Valentina, Urrutia, Alejandra, Duffy, Darragh, Cassard, Lydie, Di Santo, James P., Mottez, Estelle, Quintana-Murci, Lluis, Albert, Matthew L., Rogge, Lars, The Milieu Interieur, Consortium, Centre d'Immunologie Humaine (CIH), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche Translationnelle - Center for Translational Science (CRT), Institut Pasteur [Paris] (IP), Immunobiologie des Cellules Dendritiques, Immunité Innée - Innate Immunity, Génétique Evolutive Humaine - Human Evolutionary Genetics, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Immunorégulation, This work benefited from support of the French government's Invest in the Future Program, managed by the Agence Nationale de la Recherche (ANR, reference 10-LABX-69-01), The Milieu Intérieur Consortium is composed of the following team leaders: Laurent Abel (Hôpital Necker), Andres Alcover, Philippe Bousso, Pierre Bruhns, Ana Cumano, Marc Daëron, Cécile Delval, Caroline Demangel, Ludovic Deriano, James Di Santo, Françoise Dromer, Gérard Eberl, Jost Enninga, Antonio Freitas, Odile Gelpi, Ivo Gomperts-Boneca, Serge Hercberg (Université Paris 13), Olivier Lantz (Institut Curie), Claude Leclerc, Hugo Mouquet, Sandra Pellegrini, Stanislas Pol (Hôpital Côchin), Lars Rogge, Anavaj Sakuntabhai, Olivier Schwartz, Benno Schwikowski, Spencer Shorte, Vassili Soumelis (Institut Curie), Frédéric Tangy, Eric Tartour (Hôpital Européen George Pompidou), Antoine Toubert (Hôpital Saint-Louis), Marie-Noëlle Ungeheuer, Lluis Quintana-Murci, and Matthew L. Albert., ANR-10-LABX-0069,MILIEU INTERIEUR,GENETIC & ENVIRONMENTAL CONTROL OF IMMUNE PHENOTYPE VARIANCE: ESTABLISHING A PATH TOWARDS PERSONALIZED MEDICINE(2010), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris], Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Vougny, Marie-Christine, and Laboratoires d'excellence - GENETIC & ENVIRONMENTAL CONTROL OF IMMUNE PHENOTYPE VARIANCE: ESTABLISHING A PATH TOWARDS PERSONALIZED MEDICINE - - MILIEU INTERIEUR2010 - ANR-10-LABX-0069 - LABX - VALID
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MESH: Killer Cells, Natural ,[SDV.IMM] Life Sciences [q-bio]/Immunology ,MESH: Flow Cytometry/methods ,Neutrophils ,T-Lymphocytes ,Immunology ,Population ,Context (language use) ,MESH: Neutrophils ,MESH: Monocytes ,Monocytes ,Specimen Handling ,Immunophenotyping ,Automation ,Antigens, CD ,MESH: B-Lymphocytes ,Humans ,Immunology and Allergy ,Medicine ,education ,MESH: Automation, Laboratory/methods ,Whole blood ,Automation, Laboratory ,B-Lymphocytes ,education.field_of_study ,MESH: Humans ,Multi parametric ,MESH: Dendritic Cells ,business.industry ,MESH: Immunophenotyping/methods ,MESH: Antigens, CD/immunology ,Dendritic Cells ,Flow Cytometry ,MESH: Specimen Handling/methods ,Standardization ,3. Good health ,Killer Cells, Natural ,MESH: T-Lymphocytes ,Robotic systems ,Reference values ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,business ,Cytometry - Abstract
International audience; Immunophenotyping by multi-parametric flow cytometry is the cornerstone technology for enumeration and characterization of immune cell populations in health and disease. Standardized procedures are essential to allow for inter-individual comparisons in the context of population based or clinical studies. Herein we report the approach taken by the Milieu Intérieur Consortium, highlighting the standardized and automated procedures used for immunophenotyping of human whole blood samples. We optimized eight-color antibody panels and procedures for staining and lysis of whole blood samples, and implemented pre-analytic steps with a semi-automated workflow using a robotic system. We report on four panels that were designed to enumerate and phenotype major immune cell populations (PMN, T, B, NK cells, monocytes and DC). This work establishes a foundation for defining reference values in healthy donors. Our approach provides robust protocols for affordable, semi-automated eight-color cytometric immunophenotyping that can be used in population-based studies and clinical trial settings.
- Published
- 2015