title: Interferon regulatory factor 7 regulates airway epithelial cell responses to human rhinovirus infection.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Telethon Kids Institute, University of Western Australia, PO Box 855, West Perth, WA, 6872, Australia. Anthony.Bosco@telethonkids.org.au.
authors.forename: Anthony
authors.initials: A
authors.lastname: Bosco
authors.affiliation: Airway Inflammation Research Group, Snyder Institute for Chronic Diseases, and the Department of Physiology & Pharmacology, University of Calgary Faculty of Medicine, Calgary, AB, Canada. wiehler@ucalgary.ca.
authors.forename: Shahina
authors.initials: S
authors.lastname: Wiehler
authors.affiliation: Airway Inflammation Research Group, Snyder Institute for Chronic Diseases, and the Department of Physiology & Pharmacology, University of Calgary Faculty of Medicine, Calgary, AB, Canada. dproud@ucalgary.ca.
authors.forename: David
authors.initials: D
authors.lastname: Proud
mesh_terms: D002478:Cells, Cultured; D054365:Chemokine CXCL5 / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism* / Q000821:virology*; D006801:Humans; D050839:Interferon Regulatory Factor-7 / Q000235:genetics / Q000378:metabolism*; D000072179:Interleukin-1 Receptor-Like 1 Protein / Q000235:genetics / Q000378:metabolism; D000067596:Interleukin-33 / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D012137:Respiratory System / Q000166:cytology*; D012229:Rhinovirus / Q000502:physiology*
chemical_list: D054365:Chemokine CXCL5; C506536:IL33 protein, human; D050839:Interferon Regulatory Factor-7; D000072179:Interleukin-1 Receptor-Like 1 Protein; D000067596:Interleukin-33; D034741:RNA, Small Interfering
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