Evaluating STAT5 Phosphorylation as a Mean to Assess T Cell Proliferation

被引:33
|
作者
Bitar, Michael [1 ]
Boldt, Andreas [1 ]
Freitag, Marie-Theres [1 ]
Gruhn, Bernd [2 ]
Koehl, Ulrike [1 ,3 ,4 ]
Sack, Ulrich [1 ]
机构
[1] Univ Leipzig, Inst Clin Immunol, Med Fac, Leipzig, Germany
[2] Jena Univ Hosp, Dept Pediat, Jena, Germany
[3] Hannover Med Sch, Inst Cellular Therapeut, Hannover, Germany
[4] Fraunhofer Inst Immunol & Cell Therapy IZI, Leipzig, Germany
来源
FRONTIERS IN IMMUNOLOGY | 2019年 / 10卷
关键词
STAT5; activation; T cell proliferation; T cell activation; flow cytometry; CD25; FLOW-CYTOMETRIC MEASUREMENT; WHOLE-BLOOD; IMMUNODEFICIENCY; INTERLEUKIN-2; TRANSCRIPTION; ACTIVATION; EXPRESSION; RECEPTOR; NFAT;
D O I
10.3389/fimmu.2019.00722
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Here we present a simple and sensitive flow cytometric-based assay to assess T cell proliferation. Given the critical role STAT5A phosphorylation in T cell proliferation, we decided to evaluate phosphorylation of STAT5A as an indicator of T cell proliferation. We determined pSTAT5A in T cell treated with either CD3/CD28 or PHA. After stimulation, T cells from adult healthy donors displayed a strong long-lasting phosphorylation of STAT5A, reaching a peak value after 24 h. The median fluorescence intensity (MFI) of pSTAT5A increased from 112 +/- 17 to 512 +/- 278 (CD3/CD28) (24 h) and to 413 +/- 123 (PHA) (24 h), the IL-2 receptor-alpha (CD25) expression was greatly enhanced and after 72 h T cell proliferation amounted to 52.3 +/- 10.3% (CD3/CD28) and to 48.4 +/- 9.7% (PHA). Treatment with specific JAK3 and STAT5 inhibitors resulted in a complete blockage of phosphorylation of STAT5A, CD25 expression, and suppression of T cell proliferation. Compared with currently available methods, STAT5A phosphorylation is well-suited to predict T cell proliferation. Moreover, the method presented here is not very time consuming (several hours) and delivers functional information from which conclusions about T cell proliferation can be drawn.
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页数:11
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