title: Influenza virus damages the alveolar barrier by disrupting epithelial cell tight junctions.
abstract: A major cause of respiratory failure during influenza A virus (IAV) infection is damage to the epithelial-endothelial barrier of the pulmonary alveolus. Damage to this barrier results in flooding of the alveolar lumen with proteinaceous oedema fluid, erythrocytes and inflammatory cells. To date, the exact roles of pulmonary epithelial and endothelial cells in this process remain unclear.Here, we used an in vitro co-culture model to understand how IAV damages the pulmonary epithelial-endothelial barrier. Human epithelial cells were seeded on the upper half of a transwell membrane while human endothelial cells were seeded on the lower half. These cells were then grown in co-culture and IAV was added to the upper chamber.We showed that the addition of IAV (H1N1 and H5N1 subtypes) resulted in significant barrier damage. Interestingly, we found that, while endothelial cells mounted a pro-inflammatory/pro-coagulant response to a viral infection in the adjacent epithelial cells, damage to the alveolar epithelial-endothelial barrier occurred independently of endothelial cells. Rather, barrier damage was associated with disruption of tight junctions amongst epithelial cells, and specifically with loss of tight junction protein claudin-4.Taken together, these data suggest that maintaining epithelial cell integrity is key in reducing pulmonary oedema during IAV infection.
authors.affiliation: Dept of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands School of Biomedical Sciences, University of Queensland, Brisbane, Australia.
authors.forename: Kirsty R
authors.initials: KR
authors.lastname: Short
authors.affiliation: Institute of Pathology, University Medical Center, Johannes Gutenberg University, Mainz, Germany.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Kasper
authors.affiliation: Dept of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Stijn
authors.initials: S
authors.lastname: van der Aa
authors.affiliation: Dept of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Arno C
authors.initials: AC
authors.lastname: Andeweg
authors.affiliation: Dept of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Fatiha
authors.initials: F
authors.lastname: Zaaraoui-Boutahar
authors.affiliation: Dept of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Marco
authors.initials: M
authors.lastname: Goeijenbier
authors.affiliation: Dept of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Mathilde
authors.initials: M
authors.lastname: Richard
authors.affiliation: University of Giessen and Marburg Lung Center (UGMLC), Justus-Liebig-University of Giessen, Member of the German Center for Lung Research (DZL), Giessen, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Herold
authors.affiliation: University of Giessen and Marburg Lung Center (UGMLC), Justus-Liebig-University of Giessen, Member of the German Center for Lung Research (DZL), Giessen, Germany.
authors.forename: Christin
authors.initials: C
authors.lastname: Becker
authors.affiliation: Rocky Mountain Veterinary Branch, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA.
authors.forename: Dana P
authors.initials: DP
authors.lastname: Scott
authors.affiliation: Dept of Molecular Cell Biology, Section Electron Microscopy, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Ronald W A L
authors.initials: RW
authors.lastname: Limpens
authors.affiliation: Dept of Molecular Cell Biology, Section Electron Microscopy, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Abraham J
authors.initials: AJ
authors.lastname: Koster
authors.affiliation: Dept of Molecular Cell Biology, Section Electron Microscopy, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Montserrat
authors.initials: M
authors.lastname: Bárcena
authors.affiliation: Dept of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Ron A M
authors.initials: RA
authors.lastname: Fouchier
authors.affiliation: Institute of Pathology, University Medical Center, Johannes Gutenberg University, Mainz, Germany.
authors.forename: Charles James
authors.initials: CJ
authors.lastname: Kirkpatrick
authors.affiliation: Dept of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands t.kuiken@erasmusmc.nl.
authors.forename: Thijs
authors.initials: T
authors.lastname: Kuiken
mesh_terms: D002460:Cell Line; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D004847:Epithelial Cells / Q000473:pathology / Q000821:virology*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000472:pathogenicity*; D053124:Influenza A Virus, H5N1 Subtype / Q000472:pathogenicity*; D011650:Pulmonary Alveoli / Q000821:virology*; D019108:Tight Junctions / Q000648:ultrastructure*
chemical_list: D016207:Cytokines