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Flow Cytometric Characterization of Human Antigen-Reactive T-Helper Cells

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T-Helper Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2285))

Abstract

The detection and functional characterization of antigen-reactive T helper (Th) cells has been challenging due to their low frequency and functional heterogeneity. Antigen-reactive T cell enrichment (ARTE) allows the in-depth characterization of antigen-specific Th lymphocytes as a prerequisite for better understanding the role of adaptive immune responses in health and disease. ARTE is based on detection of the activation markers CD154 (CD40L) (expressed on all conventional Th cell subsets, Tcons) and CD137 (4-1BB) (expressed on regulatory T cells, Tregs), which are upregulated on the surface of CD4+ T cells upon short-term (7 h) in vitro stimulation with antigens in the presence of antigen-presenting cells (APCs). To substantially increase the sensitivity for the detection of antigen-specific Th cells, ARTE combines magnetic pre-enrichment of rare antigen-reactive T cells with multiparameter flow cytometry. Using CD154 and CD137 in combination allows the parallel detection of reactive Tcons and Tregs, after stimulation with the antigen. Thus, the ARTE technology now enables to characterize antigen-specific T cells with increased sensitivity of detection allowing even the investigation of antigen-specific Th cells in the naive T cell repertoire and regardless of prior knowledge of MHC alleles or antigenic epitopes.

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy (EXC 2167-390884018, Excellence Cluster Precision Medicine in Chronic Inflammation) to P.B., A.S.; DFG grant n. 433038070 to P.B., A.S.; RU5042 – miTarget to P.B., and CRC/TR 241 to A.S. The German Federal Ministry of Education and Science (BMBF)—Project InfectControl 2020 AnDiPath (Fkz 03ZZ0838B) to A.S., the European Crohn and Colitis Organisation (ECCO) to P.B.

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Authors and Affiliations

  1. Institute of Immunology, Christian-Albrechts Universität zu Kiel & Universitätsklinikum Schleswig-Holstein, Kiel, Germany

    Carina Saggau, Alexander Scheffold & Petra Bacher

  2. Institute of Clinical Molecular Biology, Christian-Albrechts Universität zu Kiel, Kiel, Germany

    Petra Bacher

Authors
  1. Carina Saggau
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  2. Alexander Scheffold
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  3. Petra Bacher
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Correspondence to Petra Bacher .

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Editors and Affiliations

  1. Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy

    Francesco Annunziato

  2. Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy

    Laura Maggi

  3. Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy

    Alessio Mazzoni

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Saggau, C., Scheffold, A., Bacher, P. (2021). Flow Cytometric Characterization of Human Antigen-Reactive T-Helper Cells. In: Annunziato, F., Maggi, L., Mazzoni, A. (eds) T-Helper Cells. Methods in Molecular Biology, vol 2285. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1311-5_12

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  • DOI: https://doi.org/10.1007/978-1-0716-1311-5_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1310-8

  • Online ISBN: 978-1-0716-1311-5

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