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.
机构:
China Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R ChinaChina Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R China
Cao Shouqiang
Wang Chao
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China Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R ChinaChina Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R China
Wang Chao
Zheng Qian
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China Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R ChinaChina Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R China
Zheng Qian
Qiao Ying
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China Med Univ, Affiliated Hosp 1, Cent Lab, Shenyang 110001, Liaoning, Peoples R ChinaChina Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R China
Qiao Ying
Xu Ke
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China Med Univ, Affiliated Hosp 1, Dept Radiol, Shenyang 110001, Liaoning, Peoples R ChinaChina Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R China
Xu Ke
Jiang Tao
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机构:
Beijing Tiantan Hosp, Dept Neurosurg, Beijing, Peoples R ChinaChina Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R China
Jiang Tao
Wu Anhua
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China Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R ChinaChina Med Univ, Hosp 1, Dept Neurosurg, Shenyang 110001, Liaoning, Peoples R China
机构:
Ludwig Boltzmann Inst Canc Res, Vienna, Austria
Univ Vet Med Vienna, Inst Anim Breeding & Genet, A-1210 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Maurer, Barbara
Nivarthi, Harini
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Austrian Acad Sci, CeMM Res Ctr Mol Med, A-1090 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Nivarthi, Harini
Prchal-Murphy, Michaela
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Univ Vet Med Vienna, Inst Pharmacol & Toxicol, A-1210 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Prchal-Murphy, Michaela
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机构:
Wingelhofer, Bettina
Chen, Doris
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Austrian Acad Sci, CeMM Res Ctr Mol Med, A-1090 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Chen, Doris
Winkler, Susanne
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Ludwig Boltzmann Inst Canc Res, Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Winkler, Susanne
Prochazkova, Jana
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Austrian Acad Sci, CeMM Res Ctr Mol Med, A-1090 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Prochazkova, Jana
Schlederer, Michaela
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Ludwig Boltzmann Inst Canc Res, Vienna, Austria
Med Univ Vienna, Dept Clin Pathol, A-1090 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Schlederer, Michaela
Pham, Ha
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Ludwig Boltzmann Inst Canc Res, Vienna, Austria
Univ Vet Med Vienna, Inst Anim Breeding & Genet, A-1210 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Pham, Ha
Loizou, Joanna I.
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Austrian Acad Sci, CeMM Res Ctr Mol Med, A-1090 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Loizou, Joanna I.
Kenner, Lukas
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Ludwig Boltzmann Inst Canc Res, Vienna, Austria
Med Univ Vienna, Dept Clin Pathol, A-1090 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Kenner, Lukas
Sexl, Veronika
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Univ Vet Med Vienna, Inst Pharmacol & Toxicol, A-1210 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Sexl, Veronika
Kolbe, Thomas
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Univ Vet Med Vienna, Biomodels Austria, A-1210 Vienna, Austria
Univ Nat Resources & Appl Life Sci, IFA Tulln, A-3430 Tulln, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Kolbe, Thomas
Kralovics, Robert
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机构:
Austrian Acad Sci, CeMM Res Ctr Mol Med, A-1090 Vienna, Austria
Univ Vet Med Vienna, Inst Lab Anim Sci, A-1210 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Kralovics, Robert
Mueller, Mathias
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机构:
Univ Vet Med Vienna, Inst Anim Breeding & Genet, A-1210 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria
Mueller, Mathias
Ruelicke, Thomas
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Univ Vet Med Vienna, Inst Lab Anim Sci, A-1210 Vienna, AustriaLudwig Boltzmann Inst Canc Res, Vienna, Austria