K Looi1,2, A G Buckley3, P J Rigby3, L W Garratt1,2, T Iosifidis1,4, G R Zosky5, A N Larcombe2,6, F J Lannigan1,7, K-M Ling2, K M Martinovich2, E Kicic-Starcevich2,8, N C Shaw2, E N Sutanto2,8, D A Knight9,10,11, A Kicic1,2,4,6,8, S M Stick1,2,4,8. 1. School of Paediatrics and Child Health, University of Western Australia, Nedlands, WA, Australia. 2. Telethon Kids Institute, University of Western Australia, Subiaco, WA, Australia. 3. Centre for Microscopy, Characterisation and Analysis (CMCA), University of Western Australia, Crawley, WA, Australia. 4. Centre for Cell Therapy and Regenerative Medicine, University of Western Australia, Nedlands, WA, Australia. 5. School of Medicine, Faculty of Health, University of Tasmania, Hohart, TAS, Australia. 6. Occupation and Environment, School of Public Health, Curtin University, Perth, WA, Australia. 7. School of Medicine, Notre Dame University, Fremantle, WA, Australia. 8. Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, WA, Australia. 9. School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, NSW, Australia. 10. Priority Research Centre for Asthma and Respiratory Disease, Hunter Medical Research Institute, Newcastle, NSW, Australia. 11. Department of Anaesthesiology, Pharmacology and Therapeutics, University of British Columbia, Vancouver, BC, Canada.
Abstract
BACKGROUND: Bronchial epithelial tight junctions (TJ) have been extensively assessed in healthy airway epithelium. However, no studies have yet assessed the effect of human rhinovirus (HRV) infection on the expression and resultant barrier function in epithelial tight junctions (TJ) in childhood asthma. OBJECTIVES: To investigate the impact of HRV infection on airway epithelial TJ expression and barrier function in airway epithelial cells (AECs) of children with and without asthma. Furthermore, to test the hypothesis that barrier integrity and function is compromised to a greater extent by HRV in AECs from asthmatic children. METHODS: Primary AECs were obtained from children with and without asthma, differentiated into air-liquid interface (ALI) cultures and infected with rhinovirus. Expression of claudin-1, occludin and zonula occluden-1 (ZO-1) was assessed via qPCR, immunocytochemistry (ICC), in-cell western (ICW) and confocal microscopy. Barrier function was assessed by transepithelial electrical resistance (TER; RT ) and permeability to fluorescent dextran. RESULTS: Basal TJ gene expression of claudin-1 and occludin was significantly upregulated in asthmatic children compared to non-asthmatics; however, no difference was seen with ZO-1. Interestingly, claudin-1, occludin and ZO-1 protein expression was significantly reduced in AEC of asthmatic children compared to non-asthmatic controls suggesting possible post-transcriptional inherent differences. HRV infection resulted in a transient dissociation of TJ and airway barrier integrity in non-asthmatic children. Although similar dissociation of TJ was observed in asthmatic children, a significant and sustained reduction in TJ expression concurrent with both a significant decrease in TER and an increase in permeability in asthmatic children was observed. CONCLUSION: This study demonstrates novel intrinsic differences in TJ gene and protein expression between AEC of children with and without asthma. Furthermore, it correlates directly the relationship between HRV infection and the resultant dissociation of epithelial TJ that causes a continued altered barrier function in children with asthma.
BACKGROUND: Bronchial epithelial tight junctions (TJ) have been extensively assessed in healthy airway epithelium. However, no studies have yet assessed the effect of human rhinovirus (HRV) infection on the expression and resultant barrier function in epithelial tight junctions (TJ) in childhood asthma. OBJECTIVES: To investigate the impact of HRV infection on airway epithelial TJ expression and barrier function in airway epithelial cells (AECs) of children with and without asthma. Furthermore, to test the hypothesis that barrier integrity and function is compromised to a greater extent by HRV in AECs from asthmatic children. METHODS: Primary AECs were obtained from children with and without asthma, differentiated into air-liquid interface (ALI) cultures and infected with rhinovirus. Expression of claudin-1, occludin and zonula occluden-1 (ZO-1) was assessed via qPCR, immunocytochemistry (ICC), in-cell western (ICW) and confocal microscopy. Barrier function was assessed by transepithelial electrical resistance (TER; RT ) and permeability to fluorescentdextran. RESULTS: Basal TJ gene expression of claudin-1 and occludin was significantly upregulated in asthmatic children compared to non-asthmatics; however, no difference was seen with ZO-1. Interestingly, claudin-1, occludin and ZO-1 protein expression was significantly reduced in AEC of asthmatic children compared to non-asthmatic controls suggesting possible post-transcriptional inherent differences. HRV infection resulted in a transient dissociation of TJ and airway barrier integrity in non-asthmatic children. Although similar dissociation of TJ was observed in asthmatic children, a significant and sustained reduction in TJ expression concurrent with both a significant decrease in TER and an increase in permeability in asthmatic children was observed. CONCLUSION: This study demonstrates novel intrinsic differences in TJ gene and protein expression between AEC of children with and without asthma. Furthermore, it correlates directly the relationship between HRV infection and the resultant dissociation of epithelial TJ that causes a continued altered barrier function in children with asthma.
Authors: W Gerald Teague; Monica G Lawrence; Debbie-Ann T Shirley; Andrea S Garrod; Stephen V Early; Jackie B Payne; Julia A Wisniewski; Peter W Heymann; James J Daniero; John W Steinke; Deborah K Froh; Thomas J Braciale; Michael Ellwood; Drew Harris; Larry Borish Journal: J Allergy Clin Immunol Pract Date: 2019-01-14
Authors: Thomas Iosifidis; Erika N Sutanto; Alysia G Buckley; Laura Coleman; Erin E Gill; Amy H Lee; Kak-Ming Ling; Jessica Hillas; Kevin Looi; Luke W Garratt; Kelly M Martinovich; Nicole C Shaw; Samuel T Montgomery; Elizabeth Kicic-Starcevich; Yuliya V Karpievitch; Peter Le Souëf; Ingrid A Laing; Shyan Vijayasekaran; Francis J Lannigan; Paul J Rigby; Robert Ew Hancock; Darryl A Knight; Stephen M Stick; Anthony Kicic Journal: JCI Insight Date: 2020-04-09
Authors: Matthew C Altman; Agustin Calatroni; Sima Ramratnam; Daniel J Jackson; Scott Presnell; Mario G Rosasco; Peter J Gergen; Leonard B Bacharier; George T O'Connor; Megan T Sandel; Meyer Kattan; Robert A Wood; Cynthia M Visness; James E Gern Journal: J Allergy Clin Immunol Date: 2021-03-10 Impact factor: 10.793
Authors: Milena Sokolowska; Valerie F J Quesniaux; Cezmi A Akdis; Kian Fan Chung; Bernhard Ryffel; Dieudonnée Togbe Journal: Front Immunol Date: 2019-09-13 Impact factor: 7.561
Authors: Christopher J Harrison; William C Weldon; Barbara A Pahud; Mary Anne Jackson; M Steven Oberste; Rangaraj Selvarangan Journal: Emerg Infect Dis Date: 2019-03 Impact factor: 6.883
Authors: S Post; I H Heijink; L Hesse; H K Koo; F Shaheen; M Fouadi; V N S Kuchibhotla; B N Lambrecht; A J M Van Oosterhout; T L Hackett; M C Nawijn Journal: Sci Rep Date: 2018-09-05 Impact factor: 4.379