title: Influenza promotes collagen deposition via αvβ6 integrin-mediated transforming growth factor β activation.
abstract: Influenza infection exacerbates chronic pulmonary diseases, including idiopathic pulmonary fibrosis. A central pathway in the pathogenesis of idiopathic pulmonary fibrosis is epithelial injury leading to activation of transforming growth factor β (TGFβ). The mechanism and functional consequences of influenza-induced activation of epithelial TGFβ are unclear. Influenza stimulates toll-like receptor 3 (TLR3), which can increase RhoA activity, a key event prior to activation of TGFβ by the αvβ6 integrin. We hypothesized that influenza would stimulate TLR3 leading to activation of latent TGFβ via αvβ6 integrin in epithelial cells. Using H1152 (IC50 6.1 μm) to inhibit Rho kinase and 6.3G9 to inhibit αvβ6 integrins, we demonstrate their involvement in influenza (A/PR/8/34 H1N1) and poly(I:C)-induced TGFβ activation. We confirm the involvement of TLR3 in this process using chloroquine (IC50 11.9 μm) and a dominant negative TLR3 construct (pZERO-hTLR3). Examination of lungs from influenza-infected mice revealed augmented levels of collagen deposition, phosphorylated Smad2/3, αvβ6 integrin, and apoptotic cells. Finally, we demonstrate that αvβ6 integrin-mediated TGFβ activity following influenza infection promotes epithelial cell death in vitro and enhanced collagen deposition in vivo and that this response is diminished in Smad3 knock-out mice. These data show that H1N1 and poly(I:C) can induce αvβ6 integrin-dependent TGFβ activity in epithelial cells via stimulation of TLR3 and suggest a novel mechanism by which influenza infection may promote collagen deposition in fibrotic lung disease.
authors.affiliation: From the Nottingham Respiratory Research Unit, University of Nottingham, Nottingham University Hospitals, Clinical Sciences Building, City Hospital Campus, Nottingham NG5 1PB, United Kingdom.
authors.forename: Lisa
authors.initials: L
authors.lastname: Jolly
authors.affiliation: From the Nottingham Respiratory Research Unit, University of Nottingham, Nottingham University Hospitals, Clinical Sciences Building, City Hospital Campus, Nottingham NG5 1PB, United Kingdom.
authors.forename: Anastasios
authors.initials: A
authors.lastname: Stavrou
authors.affiliation: the McMaster Immunology Research Centre and Firestone Institute at St. Joseph's Health Care, McMaster University, Hamilton, Ontario L8S4L8, Canada, and.
authors.forename: Gilles
authors.initials: G
authors.lastname: Vanderstoken
authors.affiliation: the Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, Tennessee 38105.
authors.forename: Victoria A
authors.initials: VA
authors.lastname: Meliopoulos
authors.affiliation: From the Nottingham Respiratory Research Unit, University of Nottingham, Nottingham University Hospitals, Clinical Sciences Building, City Hospital Campus, Nottingham NG5 1PB, United Kingdom.
authors.forename: Anthony
authors.initials: A
authors.lastname: Habgood
authors.affiliation: From the Nottingham Respiratory Research Unit, University of Nottingham, Nottingham University Hospitals, Clinical Sciences Building, City Hospital Campus, Nottingham NG5 1PB, United Kingdom.
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Tatler
authors.affiliation: From the Nottingham Respiratory Research Unit, University of Nottingham, Nottingham University Hospitals, Clinical Sciences Building, City Hospital Campus, Nottingham NG5 1PB, United Kingdom.
authors.forename: Joanne
authors.initials: J
authors.lastname: Porte
authors.affiliation: From the Nottingham Respiratory Research Unit, University of Nottingham, Nottingham University Hospitals, Clinical Sciences Building, City Hospital Campus, Nottingham NG5 1PB, United Kingdom.
authors.forename: Alan
authors.initials: A
authors.lastname: Knox
authors.affiliation: Biogen Idec Inc., Cambridge, Massachusetts 02142.
authors.forename: Paul
authors.initials: P
authors.lastname: Weinreb
authors.affiliation: Biogen Idec Inc., Cambridge, Massachusetts 02142.
authors.forename: Shelia
authors.initials: S
authors.lastname: Violette
authors.affiliation: the Manchester Collaborative Centre for Inflammation Research, University of Manchester, Manchester M13 9NT, United Kingdom.
authors.forename: Tracy
authors.initials: T
authors.lastname: Hussell
authors.affiliation: the McMaster Immunology Research Centre and Firestone Institute at St. Joseph's Health Care, McMaster University, Hamilton, Ontario L8S4L8, Canada, and.
authors.forename: Martin
authors.initials: M
authors.lastname: Kolb
authors.affiliation: the McMaster Immunology Research Centre and Firestone Institute at St. Joseph's Health Care, McMaster University, Hamilton, Ontario L8S4L8, Canada, and.
authors.forename: Martin R
authors.initials: MR
authors.lastname: Stampfli
authors.affiliation: the Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, Tennessee 38105.
authors.forename: Stacey
authors.initials: S
authors.lastname: Schultz-Cherry
authors.affiliation: From the Nottingham Respiratory Research Unit, University of Nottingham, Nottingham University Hospitals, Clinical Sciences Building, City Hospital Campus, Nottingham NG5 1PB, United Kingdom, gisli.jenkins@nottingham.ac.uk.
authors.forename: Gisli
authors.initials: G
authors.lastname: Jenkins
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism*; D000998:Antiviral Agents / Q000494:pharmacology; D017209:Apoptosis; D002461:Cell Line, Transformed; D003094:Collagen / Q000378:metabolism*; D004285:Dogs; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D015151:Immunoblotting; D053118:Influenza A Virus, H1N1 Subtype / Q000502:physiology; D016023:Integrins / Q000235:genetics / Q000378:metabolism*; D008168:Lung / Q000378:metabolism / Q000821:virology; D061985:Madin Darby Canine Kidney Cells; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009976:Orthomyxoviridae Infections / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D010766:Phosphorylation / Q000187:drug effects; D011070:Poly I-C / Q000494:pharmacology; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism; D051196:Toll-Like Receptor 3 / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: C459092:2-methyl-1-((4-methyl-5-isoquinolinyl)sulfonyl)homopiperazine; D000951:Antigens, Neoplasm; D000998:Antiviral Agents; D016023:Integrins; D051900:Smad3 Protein; D051196:Toll-Like Receptor 3; D016212:Transforming Growth Factor beta; C073878:integrin alphavbeta6; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D003094:Collagen; D054460:rho-Associated Kinases; D011070:Poly I-C
keywords: Apoptosis; Influenza; Integrin; Pulmonary Fibrosis; Toll-like Receptor (TLR)