Pseudomonas aeruginosa Exoprotein-Induced Barrier Disruption Correlates With Elastase Activity and Marks Chronic Rhinosinusitis Severity
Background:Pseudomonas aeruginosa causes severe chronic respiratory diseases and is associated with recalcitrant chronic rhinosinusitis (CRS). P. aeruginosa exoproteins contain virulence factors and play important roles in the pathogenicity of P. aeruginosa, however their role in CRS pathophysiology...
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doaj-28f9db5009794e0db98eb2ebcac5dadc2020-11-24T21:16:12ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882019-02-01910.3389/fcimb.2019.00038426718Pseudomonas aeruginosa Exoprotein-Induced Barrier Disruption Correlates With Elastase Activity and Marks Chronic Rhinosinusitis SeverityJian Li0Jian Li1Jian Li2Mahnaz Ramezanpour3Stephanie A. Fong4Clare Cooksley5Jae Murphy6Masanobu Suzuki7Masanobu Suzuki8Alkis J. Psaltis9Peter John Wormald10Sarah Vreugde11Department of Otolaryngology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, ChinaGuangzhou Key Laboratory of Otorhinolaryngology, Guangzhou, ChinaDepartment of Surgery-Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, SA, AustraliaDepartment of Otolaryngology-Head and Neck Surgery, Hokkaido University Graduate School of Medicine, Sapporo, JapanDepartment of Surgery-Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, SA, AustraliaBackground:Pseudomonas aeruginosa causes severe chronic respiratory diseases and is associated with recalcitrant chronic rhinosinusitis (CRS). P. aeruginosa exoproteins contain virulence factors and play important roles in the pathogenicity of P. aeruginosa, however their role in CRS pathophysiology remains unknown.Methods: We isolated P. aeruginosa clinical isolates (CIs) and obtained clinical information from 21 CRS patients. Elastase activity of the CIs was measured at different phases of growth. Primary human nasal epithelial cells (HNECs) were cultured at air-liquid interface (ALI) and challenged with P. aeruginosa exoproteins or purified elastase, followed by measuring Transepithelial Electrical Resistance (TEER), permeability of FITC-dextrans, western blot, and immunofluorescence.Results: 14/21 CIs had a significant increase in elastase activity in stationary phase of growth. There was a highly significant strong correlation between the in vitro elastase activity of P. aeruginosa CIs with mucosal barrier disruption evidenced by increased permeability of FITC-dextrans (r = 0.95, p = 0.0004) and decreased TEER (r = −0.9333, P < 0.01) after 4 h of challenge. Western blot showed a significant degradation of ZO-1, Occludin and β-actin in relation to the elastase activity of the exoproteins. There was a highly significant correlation between the in vitro elastase activity of P. aeruginosa CIs and CRS disease severity (for log phase, r = 0.5631, p = 0.0097; for stationary phase, r = 0.66, p = 0.0013) assessed by CT imaging of the paranasal sinuses.Conclusion: Our results implicate P. aeruginosa exoproteins as playing a major role in the pathophysiology of P. aeruginosa associated CRS by severely compromising mucosal barrier structure and function.https://www.frontiersin.org/article/10.3389/fcimb.2019.00038/fullchronic rhinosinusitisCT-scoreelastasemucosal barrierPseudomonas aeruginosa |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jian Li Jian Li Jian Li Mahnaz Ramezanpour Stephanie A. Fong Clare Cooksley Jae Murphy Masanobu Suzuki Masanobu Suzuki Alkis J. Psaltis Peter John Wormald Sarah Vreugde |
spellingShingle |
Jian Li Jian Li Jian Li Mahnaz Ramezanpour Stephanie A. Fong Clare Cooksley Jae Murphy Masanobu Suzuki Masanobu Suzuki Alkis J. Psaltis Peter John Wormald Sarah Vreugde Pseudomonas aeruginosa Exoprotein-Induced Barrier Disruption Correlates With Elastase Activity and Marks Chronic Rhinosinusitis Severity Frontiers in Cellular and Infection Microbiology chronic rhinosinusitis CT-score elastase mucosal barrier Pseudomonas aeruginosa |
author_facet |
Jian Li Jian Li Jian Li Mahnaz Ramezanpour Stephanie A. Fong Clare Cooksley Jae Murphy Masanobu Suzuki Masanobu Suzuki Alkis J. Psaltis Peter John Wormald Sarah Vreugde |
author_sort |
Jian Li |
title |
Pseudomonas aeruginosa Exoprotein-Induced Barrier Disruption Correlates With Elastase Activity and Marks Chronic Rhinosinusitis Severity |
title_short |
Pseudomonas aeruginosa Exoprotein-Induced Barrier Disruption Correlates With Elastase Activity and Marks Chronic Rhinosinusitis Severity |
title_full |
Pseudomonas aeruginosa Exoprotein-Induced Barrier Disruption Correlates With Elastase Activity and Marks Chronic Rhinosinusitis Severity |
title_fullStr |
Pseudomonas aeruginosa Exoprotein-Induced Barrier Disruption Correlates With Elastase Activity and Marks Chronic Rhinosinusitis Severity |
title_full_unstemmed |
Pseudomonas aeruginosa Exoprotein-Induced Barrier Disruption Correlates With Elastase Activity and Marks Chronic Rhinosinusitis Severity |
title_sort |
pseudomonas aeruginosa exoprotein-induced barrier disruption correlates with elastase activity and marks chronic rhinosinusitis severity |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cellular and Infection Microbiology |
issn |
2235-2988 |
publishDate |
2019-02-01 |
description |
Background:Pseudomonas aeruginosa causes severe chronic respiratory diseases and is associated with recalcitrant chronic rhinosinusitis (CRS). P. aeruginosa exoproteins contain virulence factors and play important roles in the pathogenicity of P. aeruginosa, however their role in CRS pathophysiology remains unknown.Methods: We isolated P. aeruginosa clinical isolates (CIs) and obtained clinical information from 21 CRS patients. Elastase activity of the CIs was measured at different phases of growth. Primary human nasal epithelial cells (HNECs) were cultured at air-liquid interface (ALI) and challenged with P. aeruginosa exoproteins or purified elastase, followed by measuring Transepithelial Electrical Resistance (TEER), permeability of FITC-dextrans, western blot, and immunofluorescence.Results: 14/21 CIs had a significant increase in elastase activity in stationary phase of growth. There was a highly significant strong correlation between the in vitro elastase activity of P. aeruginosa CIs with mucosal barrier disruption evidenced by increased permeability of FITC-dextrans (r = 0.95, p = 0.0004) and decreased TEER (r = −0.9333, P < 0.01) after 4 h of challenge. Western blot showed a significant degradation of ZO-1, Occludin and β-actin in relation to the elastase activity of the exoproteins. There was a highly significant correlation between the in vitro elastase activity of P. aeruginosa CIs and CRS disease severity (for log phase, r = 0.5631, p = 0.0097; for stationary phase, r = 0.66, p = 0.0013) assessed by CT imaging of the paranasal sinuses.Conclusion: Our results implicate P. aeruginosa exoproteins as playing a major role in the pathophysiology of P. aeruginosa associated CRS by severely compromising mucosal barrier structure and function. |
topic |
chronic rhinosinusitis CT-score elastase mucosal barrier Pseudomonas aeruginosa |
url |
https://www.frontiersin.org/article/10.3389/fcimb.2019.00038/full |
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