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

Bitar, Michael; Boettcher, Marcus; Boldt, Andreas; Hauck, Fabian; Koehl, Ulrike; Liebert, Uwe G.; Magg, Thomas; Schulz, Marian S. und Sack, Ulrich (2020): Flow cytometric measurement of STAT5 phosphorylation in cytomegalovirus-stimulated T cells. In: Cytometry Part A, Bd. 99, Nr. 8: S. 774-783

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.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Medizin
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• ISSN: 1552-4922
• Sprache: Englisch