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首页> 外文期刊>Immunology: An Official Journal of the British Society for Immunology >Characterization of a novel PMA-inducible pathway of interleukin-13 gene expression in T cells.
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Characterization of a novel PMA-inducible pathway of interleukin-13 gene expression in T cells.

机译：T细胞中白介素13基因表达的新型PMA诱导途径的表征。

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摘要

Although interleukin 13 (IL-13) is an important mediator of asthma and allergic diseases, the molecular mechanisms regulating IL-13 gene expression are not well understood. This study was designed to define the molecular mechanisms governing IL-13 gene expression in T cells. IL-13 expression was examined in human peripheral blood T cells and in the EL-4 T-cell line by enzyme-linked immunosorbent assay and reverse-transcription polymerase chain reaction. An IL-13 promoter deletion analysis was performed using luciferase-based reporter plasmids transiently transfected into EL-4 cells by electroporation. DNA binding factors were investigated using electrophoretic mobility shift assays. In contrast to IL-4 expression, which required concomitant activation of calcium- and protein kinase C- (PKC-) dependent signalling pathways, PKC activation alone was sufficient for IL-13 protein secretion in mitogen-primed (but not resting) peripheral blood T cells, and for IL-13 mRNA expression and promoter activity in EL-4 T cells. Promoter deletion analysis localized a phorbol 12-myristate 13-acetate (PMA)-sensitive element to a proximal promoter region between -109 and -79 base pairs upstream from the IL-13 transcription start site. This promoter region supported the binding of both constitutive and PMA-inducible nuclear factors in gel shift assays.
机译：尽管白介素13（IL-13）是哮喘和变态反应性疾病的重要介体，但调节IL-13基因表达的分子机制尚不清楚。这项研究旨在定义控制T细胞中IL-13基因表达的分子机制。通过酶联免疫吸附测定和逆转录聚合酶链反应检测人外周血T细胞和EL-4 T细胞系中的IL-13表达。使用通过电穿孔瞬时转染到EL-4细胞中的基于荧光素酶的报告质粒进行IL-13启动子缺失分析。使用电泳迁移率变动分析法研究了DNA结合因子。与需要同时激活钙和蛋白激酶C（PKC-）依赖性信号通路的IL-4表达相反，仅PKC激活就足以在有丝分裂原引发的（但不是静止的）外周血中分泌IL-13 T细胞，以及EL-4 T细胞中IL-13 mRNA的表达和启动子活性。启动子缺失分析将佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）敏感元件定位在IL-13转录起始位点上游-109和-79碱基对之间的近端启动子区域。该启动子区域支持在凝胶位移测定中组成型和PMA诱导型核因子的结合。

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