Standardized high-dimensional spectral cytometry protocol and panels for whole blood immune phenotyping in clinical and translational studies.

Autor:	Dott T; Cytometry and Biomarkers UTechS, Institut Pasteur, Université Paris Cité, Paris, France.; Translational Immunology Unit, Institut Pasteur, Université Paris Cité, Paris, France., Culina S; Cytometry and Biomarkers UTechS, Institut Pasteur, Université Paris Cité, Paris, France., Chemali R; Cytometry and Biomarkers UTechS, Institut Pasteur, Université Paris Cité, Paris, France., Mansour CA; Sony Europe BV, Surrey, UK., Dubois F; Cytometry and Biomarkers UTechS, Institut Pasteur, Université Paris Cité, Paris, France.; Translational Immunology Unit, Institut Pasteur, Université Paris Cité, Paris, France., Jagla B; Cytometry and Biomarkers UTechS, Institut Pasteur, Université Paris Cité, Paris, France.; Bioinformatics and Biostatistics Hub, Institut Pasteur, Université Paris Cité, Paris, France., Doisne JM; Innate Immunity Unit, Institut Pasteur, Université Paris Cité, Paris, France., Rogge L; Immunoregulation Unit, Institut Pasteur, Université Paris Cité, Paris, France., Huetz F; Unit of Antibodies in Therapy and Pathology, INSERM UMR1222, Institut Pasteur, Université de Paris Cité, Paris, France., Jönsson F; Unit of Antibodies in Therapy and Pathology, INSERM UMR1222, Institut Pasteur, Université de Paris Cité, Paris, France.; CNRS, Paris, France., Commere PH; Cytometry and Biomarkers UTechS, Institut Pasteur, Université Paris Cité, Paris, France., Di Santo J; Innate Immunity Unit, Institut Pasteur, Université Paris Cité, Paris, France., Terrier B; Service de Médecine Interne, Hôpital Cochin, Paris, France., Quintana-Murci L; Human Evolutionary Genetics Unit, CNRS, Institut Pasteur, Université Paris Cité, UMR2000, Paris, France., Duffy D; Cytometry and Biomarkers UTechS, Institut Pasteur, Université Paris Cité, Paris, France.; Translational Immunology Unit, Institut Pasteur, Université Paris Cité, Paris, France., Hasan M; Cytometry and Biomarkers UTechS, Institut Pasteur, Université Paris Cité, Paris, France.
Jazyk:	angličtina
Zdroj:	Cytometry. Part A : the journal of the International Society for Analytical Cytology [Cytometry A] 2024 Feb; Vol. 105 (2), pp. 124-138. Date of Electronic Publication: 2023 Oct 12.
DOI:	10.1002/cyto.a.24801
Abstrakt:	Flow cytometry is the method of choice for immunophenotyping in the context of clinical, translational, and systems immunology studies. Among the latter, the Milieu Intérieur (MI) project aims at defining the boundaries of a healthy immune response to identify determinants of immune response variation. MI used immunophenotyping of a 1000 healthy donor cohort by flow cytometry as a principal outcome for immune variance at steady state. New generation spectral cytometers now enable high-dimensional immune cell characterization from small sample volumes. Therefore, for the MI 10-year follow up study, we have developed two high-dimensional spectral flow cytometry panels for deep characterization of innate and adaptive whole blood immune cells (35 and 34 fluorescent markers, respectively). We have standardized the protocol for sample handling, staining, acquisition, and data analysis. This approach enables the reproducible quantification of over 182 immune cell phenotypes at a single site. We have applied the protocol to discern minor differences between healthy and patient samples and validated its value for application in immunomonitoring studies. Our protocol is currently used for characterization of the impact of age and environmental factors on peripheral blood immune phenotypes of >400 donors from the initial MI cohort. (© 2023 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.)
Databáze:	MEDLINE
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