Human amniotic epithelial cells inhibit CD4+T cell activation in acute kidney injury patients by influencing the miR-101-c-Rel-IL-2 pathway
Human amniotic epithelial cells inhibit CD4+T cell activation in acute kidney injury patients by influencing the miR-101-c-Rel-IL-2 pathway
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DOI：
10.1016/j.molimm.2016.11.019
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发表时间：
2017-01-01
影响因子：
3.6
通讯作者：
Xue, Jun
中科院分区：
文献类型：
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作者：
Liu, Junfeng;Hua, Rong;Xue, Jun
在急性肾损伤（阿基）的发病机制中，多种白细胞介素的释放可导致肾损伤增加。人羊膜上皮细胞（HuAECs）在体内外均能抑制免疫细胞的活化。我们假设HuAECs可以减弱患者来源的外周血CD 4 + T细胞活化并降低这些细胞表达和释放IL-2的能力。细胞增殖实验显示，在相同的培养条件下，活化的阿基患者源性CD 4 + T细胞与HuAECs共培养时增殖率显著降低。IL-2的释放水平也明显降低。Western blot和qRT-PCR检测显示，CD 4 + T细胞中c-Rel的表达也显著降低。然而，与HuAECs共培养的CD 4 + T细胞中内源性miR-101的表达水平显著增加。荧光素酶报告基因检测结果表明，miR-101可与c-Rel 3' UTR中的特定位点结合，诱导c-Rel转录后沉默。随后，我们在阿基患者来源的CD 4 + T细胞中过表达miR-101。qRT-PCR和western blot检测结果显示，内源性c-Rel的表达显著降低，而ELISA结果显示，IL-2的释放水平也显著降低。最后，ChIP-PCR检测结果显示，miR-101过表达的CD 4 + T细胞组和HuAEC共培养的CD 4 + T细胞组表现出“c-Rel-NF κ B”复合物与IL-2基因启动子之间的结合能力显著降低，并且IL-2的转录活性也显著降低。因此，我们证实HuAEC可以刺激阿基患者来源的外周血CD 4 + T细胞中的miR-101表达，从而抑制miR-101靶基因c-Rel的表达，并导致IL-2表达和释放的减少。(C)2016爱思唯尔有限公司版权所有
In the pathogenesis of acute kidney injury (AKI), the release of multiple interleukins can lead to increased kidney damage. Human amniotic epithelial cells (HuAECs) can inhibit immune cell activation in vivo and in vitro. We hypothesized that HuAECs could weaken patient-derived peripheral blood CD4+ T cell activation and decreasing the ability of these cells to express and release IL-2. Cell proliferation assay revealed that under the same culture conditions, activated AKI patient-derived CD4+ T cells had a significantly reduced proliferation rate when were co-cultured with HuAECs. And the level of IL-2 released was also significantly reduced. Western blot and qRT-PCR assays showed that the expression of c-Rel in the CD4+ T cells was also significantly reduced. However, the expression level of endogenous miR-101 in the CD4+ T cells co-cultured with HuAECs was significantly increased. Luciferase reporter assay results suggested that miR-101 could bind to a specific site in the c-Rel 3' UTR and induce the post transcriptional silencing of c-Rel. Subsequently, we over-expressed miR-101 in AKI patient-derived CD4+ T cells. The qRT-PCR and western blot assay results revealed that the expression of endogenous c-Rel was significantly reduced, while the ELISA results indicated that the level of IL-2 released was also significantly decreased. Finally, ChIP-PCR assay results showed that the miR-101-overexpressing CD4+ T-cell group and the HuAEC co-culture CD4+ T-cell group exhibited significantly decreased binding capacities between the 'c-Rel-NFKB' complex and the IL-2 gene promoter, and the transcriptional activity of IL-2 was also significantly decreased. Therefore, we confirmed that HuAECs can stimulate miR-101 expression in AKI patient-derived peripheral blood CD4+ T cells, thus inhibiting the expression of the miR-101 target gene c-Rel and leading to a reduction in IL-2 expression and release. (C) 2016 Elsevier Ltd. All rights reserved.