Mechanisms and roles by which IRF-3 mediates the regulation ofORMDL3 transcription in respiratory syncytial virus infectionXiao
Mechanisms and roles by which IRF-3 mediates the regulation ofORMDL3 transcription in respiratory syncytial virus infectionXiao
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IRF-3介导呼吸道合胞病毒感染中ORMDL3转录调控的机制及作用
DOI：
10.1016/j.biocel.2017.03.007
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
Guo-Ping Zhou
中科院分区：
文献类型：
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作者：
Xiao-Hua Wang;Jin Shu;Chun-Ming Jiang;Li-Li Zhuang;Wei-Xia Yang;Hui-Wen Zhang;Lu-Lu Wang;Lin Li;Xiao-Qing Chen;Rui Jin;Guo-Ping Zhou
呼吸道合胞病毒(RSV)是婴幼儿毛细支气管炎的主要病因，是反复喘息和哮喘的主要危险因素。据报道，OroSomucid 1样蛋白3(ORMDL3)与病毒引发的儿童反复喘息和哮喘有关。然而，关于ORMDL3是如何参与RSV感染的知之甚少。在本研究中，呼吸道合胞病毒感染毛细支气管炎患儿外周血淋巴细胞中ORMDL3的mRNA表达显著高于正常对照，体外培养的呼吸道合胞病毒感染后的支气管上皮细胞和肺成纤维细胞中ORMDL3的表达也显著增加。为了探讨RSV诱导ORMDL3表达的潜在机制，我们对ORMDL3启动子中几个转录因子的结合位点进行了电子分析。通过突变分析，近端干扰素调节因子-3(IRF-3)结合位点正向调节RSV暴露后ORMDL3的转录。针对IRF-3的过表达和RNA干扰实验表明，它调节RSV暴露后ORMDL3的表达。凝胶迁移率改变分析(EMSA)和染色质免疫沉淀(ChIP)分析表明，IRF-3与ORMDL3基因启动子直接结合。此外，我们还证实了呼吸道合胞病毒诱导的毛细支气管炎患儿外周血淋巴细胞中IRF-3的表达显著增加，并与ORMDL3的增加呈显著的线性相关。我们的结果表明，IRF-3通过直接与ORMDL3启动子结合，是RSV感染后ORMDL3诱导的重要调节因子，可能参与了毛细支气管炎和喘息性疾病的炎症和免疫反应。
Respiratory syncytial virus (RSV) is the leading cause of bronchiolitis in infancy, which is a major risk factor for recurrent wheezing and asthma. Orosomucoid 1-like protein 3 (ORMDL3) has been reported to associate with virus-triggered recurrent wheezing and asthma in children. However, little is known about howORMDL3is involved into RSV infection. In this study, we showed that the mRNA expression ofORMDL3is significantly increased in the peripheral blood lymphocytes of infants with RSV-induced bronchiolitis compared with uninfected controls, also increased in bronchial epithelial cells and lung fibroblasts following RSV infectionin vitro. To investigate the underlying mechanisms of RSV-inducedORMDL3expression, we performedin silicoanalysis of the binding sites of several transcription factors in theORMDL3promoter. The proximal interferon-regulatory factor-3 (IRF-3) binding site positively regulatedORMDL3transcription following exposure to RSV, as determined through mutational analysis. Overexpression and RNA interference experiments targeting IRF-3 showed that it regulates the expression ofORMDL3following RSV exposure. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay showed that IRF-3 binds directly to the promoter of theORMDL3gene. Furthermore, we confirmed that expression of IRF-3 is significantly increased and shows a strong linear correlation with increasedORMDL3in the peripheral blood lymphocytes from infants with RSV-induced bronchiolitis. Our results indicate that IRF-3 is an important regulator ofORMDL3induction following RSV infection by binding directly to the promoter ofORMDL3, which may be implicated in the inflammatory and immune reactions involved in bronchiolitis and wheezing diseases.