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首页> 外文期刊>Cellular immunology >Overexpressing autoimmune regulator regulates the expression of toll-like receptors by interacting with their promoters in RAW264.7 cells.
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Overexpressing autoimmune regulator regulates the expression of toll-like receptors by interacting with their promoters in RAW264.7 cells.

机译：过表达的自身免疫调节剂通过与RAW264.7细胞中的启动子相互作用来调节Toll样受体的表达。

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Autoimmune regulator (Aire) is a transcriptional activator that regulates the ectopic expression of many tissue-restricted antigens in medullary thymic epithelial cells, and that has an important role in the negative selection of autoreactive T cells. However, the roles of Aire expression in peripheral lymphoid tissues and hematopoietic cells, especially monocytes/macrophages, remain poorly understood. In this study, we found that the mRNA and protein expression levels of toll-like receptor (TLR)1, TLR3, and TLR8 were notably up-regulated in a mouse macrophage-like cell line (RAW264.7) stably expressing Aire, while the expression of TLR2, TLR4, TLR5, TLR6, TLR7, and TLR9 were not significantly changed. In addition, the mRNA expression of TLR3 and TLR8 were significantly increased in primary peritoneal macrophages transiently transfected with Aire. Using chromatin immunoprecipitation and a luciferase activity assay, we also found that Aire interacted with the TLR1, TLR3, and TLR8 promoters and increased the luciferase transcriptional activity of these promoters in RAW264.7 cells. Moreover, after stimulation by Pam(3)CSK(4), a TLR1 ligand, and poly(I:C), a TLR3 ligand, we found that the mRNA expression levels of IL-1alpha, TNFalpha, iNOS, and IFNalpha were increased in RAW264.7 cells stably expressing Aire. Together, these data suggest that Aire has a crucial role in the recognition of pathogenic microorganisms and peripheral immune tolerance in antigen-presenting cells (APCs) by regulating the expression of TLRs.
机译：自身免疫调节剂（Aire）是一种转录激活剂，可调节髓样胸腺上皮细胞中许多组织限制性抗原的异位表达，并且在负选择自身反应性T细胞中具有重要作用。然而，人们对Aire在外周淋巴组织和造血细胞，尤其是单核细胞/巨噬细胞中的作用的了解仍然很少。在这项研究中，我们发现在稳定表达Aire的小鼠巨噬细胞样细胞系（RAW264.7）中，toll​​样受体（TLR）1，TLR3和TLR8的mRNA和蛋白表达水平明显上调。 TLR2，TLR4，TLR5，TLR6，TLR7和TLR9的表达没有明显变化。此外，在用Aire瞬时转染的原代腹膜巨噬细胞中，TLR3和TLR8的mRNA表达显着增加。使用染色质免疫沉淀和萤光素酶活性测定，我们还发现Aire与TLR1，TLR3和TLR8启动子相互作用，并增加了RAW264.7细胞中这些启动子的萤光素酶转录活性。此外，在受Pam（3）CSK（4）（TLR1配体）和poly（I：C）（TLR3配体）刺激后，我们发现IL-1alpha，TNFalpha，iNOS和IFNalpha的mRNA表达水平增加了。在稳定表达Aire的RAW264.7细胞中总之，这些数据表明，Aire通过调节TLRs的表达在识别抗原呈递细胞（APC）中的病原微生物和外周免疫耐受中具有至关重要的作用。

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