Flow cytometric measurement of STAT5 phosphorylation in cytomegalovirus-stimulated T cells
| Autor: | Marcus Boettcher, Fabian Hauck, Michael Bitar, Andreas Boldt, Ulrike Köhl, Uwe G. Liebert, Thomas Magg, Marian Simon Schulz, Ulrich Sack |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Histology T cell T-Lymphocytes Congenital cytomegalovirus infection Cytomegalovirus Stimulation CD8-Positive T-Lymphocytes medicine.disease_cause Pathology and Forensic Medicine STAT5A Flow cytometry Viral Matrix Proteins 03 medical and health sciences 0302 clinical medicine medicine STAT5 Transcription Factor Humans Phosphorylation STAT5 Mutation biology medicine.diagnostic_test Chemistry virus diseases Cell Biology medicine.disease Phosphoproteins Molecular biology 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis Cytomegalovirus Infections biology.protein |
| Zdroj: | Cytometry. Part A : the journal of the International Society for Analytical CytologyREFERENCES. 99(8) |
| ISSN: | 1552-4930 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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