Granulysin production and anticryptococcal activity is dependent upon a far upstream enhancer that binds STAT5 in human peripheral blood CD4+ T cells.
Granulysin production and anticryptococcal activity is dependent upon a far upstream enhancer that binds STAT5 in human peripheral blood CD4+ T cells.
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DOI：
10.4049/jimmunol.1001725
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发表时间：
2010-11-01
期刊：
影响因子：
--
通讯作者：
Zheng C
中科院分区：
文献类型：
--
作者：
Xing J;Wu F;Wang S;Krensky AM;Mody CH;Zheng C
先前的研究已经证明STAT 5对于颗粒溶解素的表达和抗微生物活性是关键的。由于HIV感染患者的信号通路和由此产生的杀微生物活性是有缺陷的，因此STAT 5导致颗粒溶解素表达的机制引起了极大的兴趣。在目前的研究中，IL-2刺激的CRL-2105 CD 4 + T细胞表达颗粒溶素并杀死新生隐球菌，与原代CD 4 + T细胞相似。在原代CD 4 + T细胞和CRL-2105 T细胞中用荧光素酶报告基因测定分析颗粒溶解素启动子上游元件的增强子活性，并将转录起始位点上游18，302至18，177 bp的STAT 5结合位点鉴定为增强子。此外，增强子在异源SV 40启动子的背景下起作用，而不管其转录方向如何。染色质免疫沉淀和EMSA表明，增强子元件结合STAT 5在体内和体外，和STAT 5结合位点的突变废除其增强子活性。此外，显性负性STAT 5a的过表达废除了STAT 5结合位点的增强子活性，并废除了IL-2刺激的原代CD 4 + T细胞的抗隐球菌活性。总之，这些数据提供了有关导致颗粒溶素表达和抗隐球菌活性的原代CD 4 + T细胞的复杂调节的细节。
Previous studies have demonstrated that STAT5 is critical for expression of granulysin and antimicrobial activity. Because the signaling pathway and the resultant microbicidal activity are defective in HIV-infected patients, the mechanism by which STAT5 leads to granulysin expression is of great interest. In the current study, IL-2–stimulated CRL-2105 CD4+ T cells expressed granulysin and killed Cryptococcus neoformans similar to primary CD4+ T cells. The enhancer activity of the upstream element of the granulysin promoter was analyzed in primary CD4+ T cells and CRL-2105 T cells with a luciferase reporter assay, and a STAT5 binding site, 18,302 to 18,177 bp upstream of the transcription start site, was identified as an enhancer. Additionally, the enhancer functioned in the context of heterologous SV40 promoter irrespective of its transcriptional orientation. Chromatin immunoprecipitation and EMSAs demonstrated that the enhancer element bound STAT5 both in vivo and in vitro, and mutation of the STAT5 binding site abrogated its enhancer activity. Furthermore, overexpression of a dominant negative STAT5a abolished the enhancer activity of the STAT5 binding site and abrogated the anticryptococcal activity of IL-2–stimulated primary CD4+ T cells. Taken together, these data provide details about the complex regulation leading to granulysin expression and anticryptococcal activity in primary CD4+ T cells.
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影响因子：
15.3
作者：
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影响因子：
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