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  4. Flow cytometric measurement of STAT5 phosphorylation in cytomegalovirus-stimulated T cells
 
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2021
Journal Article
Title

Flow cytometric measurement of STAT5 phosphorylation in cytomegalovirus-stimulated T cells

Abstract
Cytomegalovirus (CMV)-specific T cells expand with CMV reactivation and are probably prerequisite for control and protection. Given the critical role STAT5A phosphorylation (pSTAT5A) in T cell proliferation, this study presents a simple and sensitive flow cytometric-based pSTAT5A assay to quickly identify CMV-specific T cell proliferation. We determined pSTAT5A in T cells treated with CMV-specific peptide mix (pp65 + IE1 peptides) from 20 healthy adult subjects and three immunodeficient patients with CARMIL-2 mutation. After stimulation, the percentage of pSTAT5A+ T cells in CMV-seropositive (CMV+) subjects significantly increased from 3.0% ± 1.9% (unstimulated) to 11.4% ± 5.9% (stimulated) for 24 h. After 7 days of stimulation, the percentage of expanded T cells amounted to 26% ± 17.2%. Conversely, the percentage of pSTAT5A+ T cells and T cell proliferation from CMV-seronegative (CMV-) subjects hardly changed (from 3.0% ± 1.3% to 3.7% ± 1.8% and from 4.3% ± 2.1% to 5.7% ± 1.7%, respectively). We analyzed the correlation between the percentage of pSTAT5A+ T cells versus (1) CMV-IgG concentrations versus (2) the percentage of expanded T cells and versus (3) the percentage of initial CMV-specific T cells. In immunodeficient patients with CARMIL-2 mutation, CMV-specific pSTAT5A and T cell proliferation were completely deficient. In conclusion, flow cytometric-based pSTAT5A assay represents an appropriate tool to quickly identify CMV-specific T cell proliferation and helps to understand dysfunctions in controlling other pathogens. Flow cytometric-based pSTAT5A assay may be a useful test in clinical practice and merits further validation in large studies.
Author(s)
Bitar, Michael
Universität Leipzig
Boettcher, Michael
Universität Leipzig
Boldt, Andreas
Universität Leipzig
Hauck, Fabian
LMU Klinikum München
Koehl, Ulrike
Fraunhofer-Institut für Zelltherapie und Immunologie IZI  
Liebert, Uwe G.
Universität Leipzig
Magg, Thomas
LMU Klinikum München
Schulz, Marian S.
Universität Leipzig
Sack, Ulrich
Universität Leipzig
Journal
Cytometry. Part A  
Open Access
DOI
10.1002/cyto.a.24286
Additional full text version
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Language
English
Fraunhofer-Institut für Zelltherapie und Immunologie IZI  
Keyword(s)
  • Bibliographie

  • CMV-specific T cells

  • CMV T cell proliferation

  • Durchflusszytometrie

  • STAT5 phosphorylation

  • T-Zell-Aktivierung

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