Interferon regulatory factor 7 regulates airway epithelial cell responses to human rhinovirus infection.
Interferon regulatory factor 7 regulates airway epithelial cell responses to human rhinovirus infection.
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DOI：
10.1186/s12864-016-2405-z
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发表时间：
2016-01-25
期刊：
影响因子：
4.4
通讯作者：
Proud D
中科院分区：
文献类型：
--
作者：
Bosco A;Wiehler S;Proud D
人鼻病毒（HRV）引起大多数感冒，并引发慢性下呼吸道疾病的恶化。呼吸道上皮细胞是HRV感染和复制的主要部位，也是宿主炎症反应的起始部位。目前，支持呼吸道上皮细胞HRV反应的分子机制还不完全清楚。本研究的目的是采用微阵列分析，上游调控分析，和siRNA介导的基因沉默，以进一步了解干扰素调节因子7（IRF7）在这种反应中的作用。使用Lipofectamine用靶向IRF7的siRNA或非沉默对照（全明星对照）转染原代人支气管上皮细胞（HBE）。使细胞恢复，然后在存在或不存在HRV-16的情况下培养24小时。基因表达的总体模式在微阵列上进行了分析。使用真实的时间RT-qPCR、ELISA和蛋白质印迹在mRNA和/或蛋白质水平验证微阵列研究中鉴定的基因子集。在HRV感染过程中，HBE中有数百个基因表达上调。通路分析表明这些基因主要参与I型和II型干扰素信号转导、RIG-I/MDA5信号转导、抗原加工和呈递以及细胞凋亡。这些数据的上游调节分析表明，IRF7是这种反应的主要分子驱动因素。IRF7的敲低降低了参与抗病毒应答的基因（干扰素信号传导、Toll样受体信号传导、NOD样受体信号传导、RIG-I/MDA5信号传导）的HRV驱动的上调，并增加了促进炎症（例如CXCL5、IL-33、IL1RL1）和对氧化应激的应答的基因的表达。然而，在HBE细胞中被HRV干扰的大多数基因，包括那些已知由IRF7调节的基因，对IRF7敲低不敏感。对IRF7敲低不敏感的反应部分的上游调节子分析表明，它是由NF-κ B、STAT 1、STAT 3和IRF1驱动的。我们的研究结果表明，IRF7调节参与抗病毒免疫，炎症，并在HRV感染HBE细胞的氧化应激反应的基因的表达，也表明，其他转录因子在这种反应中发挥了重要作用。本文的在线版本（doi：10.1186/s12864 - 016 - 2405-z）包含补充材料，可供授权用户使用。
Human rhinoviruses (HRV) cause the majority of colds and trigger exacerbations of chronic lower airway diseases. Airway epithelial cells are the primary site for HRV infection and replication, and the initiation of host inflammatory responses. At present, the molecular mechanisms that underpin HRV responses in airway epithelial cells are incompletely understood. The aim of this study was to employ microarray profiling, upstream regulator analysis, and siRNA mediated gene silencing to further our understanding of the role of interferon regulatory factor 7 (IRF7) in this response. Primary human bronchial epithelial cells (HBE) where transfected with siRNA that targets IRF7 or a non-silencing control (all-star control) using Lipofectamine. The cells were allowed to recover, and then cultured in the presence or absence of HRV-16 for 24 h. Global patterns of gene expression were profiled on microarrays. A subset of genes identified in the microarray study were validated at the mRNA and/or protein level using real time RT-qPCR, ELISA, and western blots. Hundreds of genes were upregulated in HBE during HRV infection. Pathways analysis demonstrated that these genes were mainly involved in type I and II interferon signaling, RIG-I/MDA5 signaling, antigen processing and presentation, and apoptosis. Upstream regulator analysis of these data suggested that IRF7 was a major molecular driver of this response. Knockdown of IRF7 reduced the HRV-driven upregulation of genes involved in antiviral responses (interferon signaling, Toll-like receptor signaling, NOD-like receptor signaling, RIG-I/MDA5 signaling), and increased the expression of genes that promote inflammation (e.g. CXCL5, IL-33, IL1RL1) and the response to oxidative stress. However, the majority of genes that were perturbed by HRV in HBE cells including those that are known to be regulated by IRF7 were insensitive to IRF7 knockdown. Upstream regulator analysis of the part of the response that was insensitive to IRF7 knockdown suggested it was driven by NF-κB, STAT1, STAT3, and IRF1. Our findings demonstrate that IRF7 regulates the expression of genes involved in antiviral immunity, inflammation, and the response to oxidative stress during HRV infections in HBE cells, and also suggests that other transcription factors play a major role in this response. The online version of this article (doi:10.1186/s12864-016-2405-z) contains supplementary material, which is available to authorized users.