title: Effect of human rhinovirus infection on airway epithelium tight junction protein disassembly and transepithelial permeability.
abstract: No studies have assessed the effects of human rhinovirus (HRV) infection on epithelial tight junctions (TJs) and resultant barrier function.
  
  
  
  To correlate viral infection with TJ disassembly, epithelial barrier integrity, and function.
  
  
  
  Human airway epithelial cells were infected with HRV minor serotype 1B (HRV-1B) at various 50% tissue culture infectivity doses (TCID50) over 72 hours. HRV replication was assessed by quantitative-polymerase chain reaction (qPCR) while cell viability and apoptosis were assessed by proliferation and apoptotic assays, respectively. Protein expression of claudin-1, occludin, and zonula occludens protein-1 (ZO-1) was assessed using In-Cell™ Western assays. Transepithelial permeability assays were performed to assess effects on barrier functionality. RT2Profiler focused qPCR arrays and pathway analysis evaluating associations between human TJ and antiviral response were performed to identify potential interactions and pathways between genes of interests.
  
  
  
  HRV-1B infection affected viability that was both time and TCID50dependent. Significant increases in apoptosis and viral replication post-infection correlated with viral titer. Viral infection significantly decreased claudin-1 protein expression at the lower TCID50, while a significant decrease in all three TJ protein expressions occurred at higher TCID50. Decrease in protein expression was concomitant with significant increases in epithelial permeability of fluorescein isothiocynate labeled-dextran 4 and 20 kDa. Analysis of focused qPCR arrays demonstrated a significant decrease in ZO-1 gene expression. Furthermore, network analysis between human TJ and antiviral response genes revealed possible interactions and regulation of TJ genes via interleukin (IL)-15 in response to HRV-1B infection.
  
  
  
  HRV-1B infection directly alters human airway epithelial TJ expression leading to increased epithelial permeability potentially via an antiviral response of IL-15.
authors.affiliation: a School of Paediatrics and Child Health , The University of Western Australia , Nedlands , Western Australia , Australia.
authors.forename: Kevin
authors.initials: K
authors.lastname: Looi
authors.affiliation: b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Niamh M
authors.initials: NM
authors.lastname: Troy
authors.affiliation: a School of Paediatrics and Child Health , The University of Western Australia , Nedlands , Western Australia , Australia.|b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Luke W
authors.initials: LW
authors.lastname: Garratt
authors.affiliation: a School of Paediatrics and Child Health , The University of Western Australia , Nedlands , Western Australia , Australia.|c Centre for Cell Therapy and Regenerative Medicine , School of Medicine and Pharmacology, The University of Western Australia , Nedlands , Western Australia , Australia.
authors.forename: Thomas
authors.initials: T
authors.lastname: Iosifidis
authors.affiliation: b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Anthony
authors.initials: A
authors.lastname: Bosco
authors.affiliation: d Centre for Microscopy, Characterisation and Analysis , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Alysia G
authors.initials: AG
authors.lastname: Buckley
authors.affiliation: b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Kak-Ming
authors.initials: KM
authors.lastname: Ling
authors.affiliation: b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Martinovich
authors.affiliation: b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Kicic-Starcevich
authors.affiliation: b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Nicole C
authors.initials: NC
authors.lastname: Shaw
authors.affiliation: b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.|e Department of Respiratory Medicine , Princess Margaret Hospital for Children , Perth , Western Australia , Australia.
authors.forename: Erika N
authors.initials: EN
authors.lastname: Sutanto
authors.affiliation: f School of Medicine, Faculty of Health , University of Tasmania , Hobart , Tasmania , Australia.
authors.forename: Graeme R
authors.initials: GR
authors.lastname: Zosky
authors.affiliation: d Centre for Microscopy, Characterisation and Analysis , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Rigby
authors.affiliation: b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.
authors.forename: Alexander N
authors.initials: AN
authors.lastname: Larcombe
authors.affiliation: g School of Biomedical Sciences and Pharmacy , University of Newcastle , Callaghan , New South Wales , Australia.|h Priority Research Centre for Asthma and Respiratory Disease , Hunter Medical Research Institute , Newcastle , New South Wales , Australia.|i Department of Anesthesiology , Pharmacology and Therapeutics, University of British Columbia , Vancouver , Canada.
authors.forename: Darryl A
authors.initials: DA
authors.lastname: Knight
authors.affiliation: a School of Paediatrics and Child Health , The University of Western Australia , Nedlands , Western Australia , Australia.|b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.|c Centre for Cell Therapy and Regenerative Medicine , School of Medicine and Pharmacology, The University of Western Australia , Nedlands , Western Australia , Australia.|e Department of Respiratory Medicine , Princess Margaret Hospital for Children , Perth , Western Australia , Australia.
authors.forename: Anthony
authors.initials: A
authors.lastname: Kicic
authors.affiliation: a School of Paediatrics and Child Health , The University of Western Australia , Nedlands , Western Australia , Australia.|b Telethon Kids Institute, Centre for Health Research , The University of Western Australia , Crawley , Western Australia , Australia.|c Centre for Cell Therapy and Regenerative Medicine , School of Medicine and Pharmacology, The University of Western Australia , Nedlands , Western Australia , Australia.|e Department of Respiratory Medicine , Princess Margaret Hospital for Children , Perth , Western Australia , Australia.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Stick
keywords: airway epithelial cells; epithelial barrier; human rhinovirus; tight junctions