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Discrimination of T-cell subsets and T-cell receptor repertoire distribution

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Abstract

Flow cytometry-based analysis of T-cell receptor (TCR) repertoires is an essential tool for the detection of clonal T-cell expansions in physiologic and pathologic conditions. Individual T-cell subsets can be investigated based on their surface properties. The aims of our study were to provide reference values for various disease settings and delineate the contribution of individual TCR repertoires to the human T-cell differentiation pathway. We analyzed blood of 66 healthy subjects aged 0 (cord blood) to 72 years. Lymphocyte subpopulations and TCR repertoires were simultaneously explored using antibodies specific to CD3, CD4, CD8, CD45RA, CCR7, CD27, CD57 and a set of 25 antibodies detecting human TCR-Vβ chains. Statistical analysis included Wilcoxon, paired t and ANOVA tests. Initially, TCR expansion values were calculated based on the analysis of TCR-Vβ distribution on CD4+ and CD8+ T cells. We then established gating strategies and an algorithm for data analysis allowing for discrimination of T-cell subsets and TCR distribution. Dominant TCR expansions were present within effector as opposed to central/effector memory or naive cells, e.g., median TCR-Vβ expansion rate was highest on CD45RA+/CCR7− effector CD4+/8+ cells (eight and 11-fold, respectively). Remarkably, TCR expansions were missing (0) or very low (0.5) on CD4+ and CD8+ central memory population, respectively. No significant gender-related variability of TCR repertoires was identified, and significant impact of chronic cytomegalovirus infection was shown. Our results serve as reference for future studies elucidating clonal TCR dominance of T-cell subsets.

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Authors and Affiliations

  1. Institute of Medical Immunology, Charité-University Medicine, Campus Virchow-Klinikum, Institutsgebäude Süd, Föhrer Str. 15/Südstr. 2, 13353, Berlin, Germany

    Isabell Bretschneider, Christian Meisel, Manuel Guerreiro, Mathias Streitz & Hans-Dieter Volk

  2. Clinic and Polyclinic for Pediatrics and Adolescent Medicine, University Hospital Carl Gustav Carus, Technical University Dresden, Dresden, Germany

    Isabell Bretschneider

  3. Translational Hematology Oncology Research, Cleveland Clinic, Cleveland, OH, USA

    Michael J. Clemente, Jaroslav P. Maciejewski & Marcin W. Wlodarski

  4. Institute of Biometrics and Medical Statistics, Charité-University Medicine, Berlin, Germany

    Werner Hopfenmüller

  5. Berlin Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany

    Christian Meisel, Mathias Streitz & Hans-Dieter Volk

  6. Department of Pediatric Hematology and Oncology, University of Freiburg, Freiburg, Germany

    Marcin W. Wlodarski

Authors
  1. Isabell Bretschneider
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  2. Michael J. Clemente
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  3. Christian Meisel
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  4. Manuel Guerreiro
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  5. Mathias Streitz
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  6. Werner Hopfenmüller
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  7. Jaroslav P. Maciejewski
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  8. Marcin W. Wlodarski
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  9. Hans-Dieter Volk
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Correspondence to Isabell Bretschneider.

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Bretschneider, I., Clemente, M.J., Meisel, C. et al. Discrimination of T-cell subsets and T-cell receptor repertoire distribution. Immunol Res 58, 20–27 (2014). https://doi.org/10.1007/s12026-013-8473-0

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  • DOI: https://doi.org/10.1007/s12026-013-8473-0

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