{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,8,30]],"date-time":"2023-08-30T13:13:33Z","timestamp":1693401213228},"reference-count":35,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2020,12,20]],"date-time":"2020-12-20T00:00:00Z","timestamp":1608422400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2021,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Cytomegalovirus (CMV)\u2010specific 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\u2010based pSTAT5A assay to quickly identify CMV\u2010specific T cell proliferation. We determined pSTAT5A in T cells treated with CMV\u2010specific peptide mix (pp65\u2009+\u2009IE1 peptides) from 20 healthy adult subjects and three immunodeficient patients with <jats:italic>CARMIL<\/jats:italic>\u20102 mutation. After stimulation, the percentage of pSTAT5A<jats:sup>+<\/jats:sup> T cells in CMV\u2010seropositive (CMV<jats:sup>+<\/jats:sup>) subjects significantly increased from 3.0%\u2009\u00b1\u20091.9% (unstimulated) to 11.4%\u2009\u00b1\u20095.9% (stimulated) for 24\u2009h. After 7\u2009days of stimulation, the percentage of expanded T cells amounted to 26%\u2009\u00b1\u200917.2%. Conversely, the percentage of pSTAT5A<jats:sup>+<\/jats:sup> T cells and T cell proliferation from CMV\u2010seronegative (CMV<jats:sup>\u2212<\/jats:sup>) subjects hardly changed (from 3.0%\u2009\u00b1\u20091.3% to 3.7%\u2009\u00b1\u20091.8% and from 4.3%\u2009\u00b1\u20092.1% to 5.7%\u2009\u00b1\u20091.7%, respectively). We analyzed the correlation between the percentage of pSTAT5A<jats:sup>+<\/jats:sup> T cells versus (1) CMV\u2010IgG concentrations versus (2) the percentage of expanded T cells and versus (3) the percentage of initial CMV\u2010specific T cells. In immunodeficient patients with <jats:italic>CARMIL\u20102<\/jats:italic> mutation, CMV\u2010specific pSTAT5A and T cell proliferation were completely deficient. In conclusion, flow cytometric\u2010based pSTAT5A assay represents an appropriate tool to quickly identify CMV\u2010specific T cell proliferation and helps to understand dysfunctions in controlling other pathogens. Flow cytometric\u2010based pSTAT5A assay may be a useful test in clinical practice and merits further validation in large studies.<\/jats:p>","DOI":"10.1002\/cyto.a.24286","type":"journal-article","created":{"date-parts":[[2020,12,6]],"date-time":"2020-12-06T16:26:05Z","timestamp":1607271965000},"page":"774-783","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Flow cytometric measurement of <scp>STAT5<\/scp> phosphorylation in cytomegalovirus\u2010stimulated T cells"],"prefix":"10.1002","volume":"99","author":[{"given":"Michael","family":"Bitar","sequence":"first","affiliation":[{"name":"Institute of Clinical Immunology, Medical Faculty University of Leipzig  Leipzig Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marcus","family":"Boettcher","sequence":"additional","affiliation":[{"name":"Institute of Clinical Immunology, Medical Faculty University of Leipzig  Leipzig 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