Effects of human rhinovirus on epithelial barrier integrity and function in children with asthma

K. Looi, A. G. Buckley, P. J. Rigby, L. W. Garratt, T. Iosifidis, G. R. Zosky, A. N. Larcombe, F. J. Lannigan, K. M. Ling, K. M. Martinovich, E. Kicic-Starcevich, N. C. Shaw, E. N. Sutanto, D. A. Knight, A. Kicic, S. M. Stick

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)513-524
Number of pages12
JournalClinical and Experimental Allergy
Volume48
Issue number5
DOIs
Publication statusPublished - 1 May 2018

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Rhinovirus
Tight Junctions
Asthma
Claudin-1
Occludin
Epithelial Cells
Infection
Permeability
Zonula Occludens-1 Protein
Tight Junction Proteins
Gene Expression
Dextrans
Electric Impedance
Confocal Microscopy
Epithelium
Immunohistochemistry
Air

Cite this

Looi, K. ; Buckley, A. G. ; Rigby, P. J. ; Garratt, L. W. ; Iosifidis, T. ; Zosky, G. R. ; Larcombe, A. N. ; Lannigan, F. J. ; Ling, K. M. ; Martinovich, K. M. ; Kicic-Starcevich, E. ; Shaw, N. C. ; Sutanto, E. N. ; Knight, D. A. ; Kicic, A. ; Stick, S. M. / Effects of human rhinovirus on epithelial barrier integrity and function in children with asthma. In: Clinical and Experimental Allergy. 2018 ; Vol. 48, No. 5. pp. 513-524.
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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.",
keywords = "asthma, epithelial permeability, human rhinovirus, tight junction",
author = "K. Looi and Buckley, {A. G.} and Rigby, {P. J.} and Garratt, {L. W.} and T. Iosifidis and Zosky, {G. R.} and Larcombe, {A. N.} and Lannigan, {F. J.} and Ling, {K. M.} and Martinovich, {K. M.} and E. Kicic-Starcevich and Shaw, {N. C.} and Sutanto, {E. N.} and Knight, {D. A.} and A. Kicic and Stick, {S. M.}",
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Looi, K, Buckley, AG, Rigby, PJ, Garratt, LW, Iosifidis, T, Zosky, GR, Larcombe, AN, Lannigan, FJ, Ling, KM, Martinovich, KM, Kicic-Starcevich, E, Shaw, NC, Sutanto, EN, Knight, DA, Kicic, A & Stick, SM 2018, 'Effects of human rhinovirus on epithelial barrier integrity and function in children with asthma' Clinical and Experimental Allergy, vol. 48, no. 5, pp. 513-524. https://doi.org/10.1111/cea.13097

Effects of human rhinovirus on epithelial barrier integrity and function in children with asthma. / Looi, K.; Buckley, A. G.; Rigby, P. J.; Garratt, L. W.; Iosifidis, T.; Zosky, G. R.; Larcombe, A. N.; Lannigan, F. J.; Ling, K. M.; Martinovich, K. M.; Kicic-Starcevich, E.; Shaw, N. C.; Sutanto, E. N.; Knight, D. A.; Kicic, A.; Stick, S. M.

In: Clinical and Experimental Allergy, Vol. 48, No. 5, 01.05.2018, p. 513-524.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effects of human rhinovirus on epithelial barrier integrity and function in children with asthma

AU - Looi, K.

AU - Buckley, A. G.

AU - Rigby, P. J.

AU - Garratt, L. W.

AU - Iosifidis, T.

AU - Zosky, G. R.

AU - Larcombe, A. N.

AU - Lannigan, F. J.

AU - Ling, K. M.

AU - Martinovich, K. M.

AU - Kicic-Starcevich, E.

AU - Shaw, N. C.

AU - Sutanto, E. N.

AU - Knight, D. A.

AU - Kicic, A.

AU - Stick, S. M.

PY - 2018/5/1

Y1 - 2018/5/1

N2 - 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.

AB - 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.

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KW - epithelial permeability

KW - human rhinovirus

KW - tight junction

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