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Effects of human neutrophil elastase and Pseudomonas aeruginosa proteinases on human respiratory epithelium

Effects of human neutrophil elastase and Pseudomonas aeruginosa proteinases on human respiratory epithelium
Effects of human neutrophil elastase and Pseudomonas aeruginosa proteinases on human respiratory epithelium

It has been suggested that proteinase enzymes could play an important role in the pathogenesis of chronic bronchial infections including bronchiectasis and cystic fibrosis (CF). Because Pseudomonas aeruginosa frequently colonizes the respiratory tract in bronchiectasis and CF, we examined the in vitro effects of human neutrophil elastase (HNE) and proteinase enzymes produced by P. aeruginosa (elastase: PE; alkaline proteinase: PAP) on the ciliary beat frequency (CBF) and ultrastructure of human nasal ciliated respiratory epithelium. HNE (500 micrograms/ml) progressively reduced CBF and caused marked epithelial disruption; lower concentrations (100 and 20 micrograms/ml) also caused epithelial disruption but without slowing CBF. The effects of HNE (500 micrograms/ml) were completely abolished by adding alpha 1-antitrypsin (5 mg/ml). There was no synergy between HNE and pyocyanin, a product of P. aeruginosa which slows CBF. PE in phosphate-buffered saline also caused epithelial disruption without slowing CBF; however, PE in medium containing divalent metal ions caused CBF slowing as well as epithelial disruption at 100 micrograms/ml. PAP (500 micrograms/ml) had almost no effect on ciliated epithelium. The effects of HNE and PE on nasal and bronchial epithelium obtained from the same patient were similar. Light and transmission electron microscopy revealed that HNE and PE were cytotoxic and caused detachment of epithelial cells from neighboring cells and the basement membrane. There was cytoplasmic blebbing of the cell surface and mitochondrial damage; however, no increase of abnormalities in the ultrastructure of cilia on living cells was seen. These results support the hypothesis that HNE and PE contribute to the delayed mucociliary clearance and epithelial damage that is observed in patients with chronic bronchial infection.

Bronchi, Cilia, Endopeptidases, Epithelium, Humans, Microscopy, Electron, Nasal Mucosa, Neutrophils, Pancreatic Elastase, Pseudomonas aeruginosa, Pyocyanine, alpha 1-Antitrypsin, Journal Article, Research Support, Non-U.S. Gov't
1044-1549
26-32
Amitani, R.
8d19850e-0227-4cf2-9f19-10334eda62a7
Wilson, R.
94eba368-b33f-4071-beb6-dac98ef23651
Rutman, A.
b06264ba-bff8-4e7b-b3af-868cfe6369e6
Read, R.
b5caca7b-0063-438a-b703-7ecbb6fc2b51
Ward, C.
49730117-7b7d-4410-b590-0314cb81d0d7
Burnett, D.
fc246532-53d1-47e2-b1da-358897c0906e
Stockley, R.A.
d3f106d5-cab5-49fb-92d7-1215c0d65823
Cole, P.J.
8d93f747-e6ed-4557-89ea-e12e0925bbf3
Amitani, R.
8d19850e-0227-4cf2-9f19-10334eda62a7
Wilson, R.
94eba368-b33f-4071-beb6-dac98ef23651
Rutman, A.
b06264ba-bff8-4e7b-b3af-868cfe6369e6
Read, R.
b5caca7b-0063-438a-b703-7ecbb6fc2b51
Ward, C.
49730117-7b7d-4410-b590-0314cb81d0d7
Burnett, D.
fc246532-53d1-47e2-b1da-358897c0906e
Stockley, R.A.
d3f106d5-cab5-49fb-92d7-1215c0d65823
Cole, P.J.
8d93f747-e6ed-4557-89ea-e12e0925bbf3

Amitani, R., Wilson, R., Rutman, A., Read, R., Ward, C., Burnett, D., Stockley, R.A. and Cole, P.J. (1991) Effects of human neutrophil elastase and Pseudomonas aeruginosa proteinases on human respiratory epithelium. American Journal of Respiratory Cell and Molecular Biology, 4 (1), 26-32. (doi:10.1165/ajrcmb/4.1.26).

Record type: Article

Abstract

It has been suggested that proteinase enzymes could play an important role in the pathogenesis of chronic bronchial infections including bronchiectasis and cystic fibrosis (CF). Because Pseudomonas aeruginosa frequently colonizes the respiratory tract in bronchiectasis and CF, we examined the in vitro effects of human neutrophil elastase (HNE) and proteinase enzymes produced by P. aeruginosa (elastase: PE; alkaline proteinase: PAP) on the ciliary beat frequency (CBF) and ultrastructure of human nasal ciliated respiratory epithelium. HNE (500 micrograms/ml) progressively reduced CBF and caused marked epithelial disruption; lower concentrations (100 and 20 micrograms/ml) also caused epithelial disruption but without slowing CBF. The effects of HNE (500 micrograms/ml) were completely abolished by adding alpha 1-antitrypsin (5 mg/ml). There was no synergy between HNE and pyocyanin, a product of P. aeruginosa which slows CBF. PE in phosphate-buffered saline also caused epithelial disruption without slowing CBF; however, PE in medium containing divalent metal ions caused CBF slowing as well as epithelial disruption at 100 micrograms/ml. PAP (500 micrograms/ml) had almost no effect on ciliated epithelium. The effects of HNE and PE on nasal and bronchial epithelium obtained from the same patient were similar. Light and transmission electron microscopy revealed that HNE and PE were cytotoxic and caused detachment of epithelial cells from neighboring cells and the basement membrane. There was cytoplasmic blebbing of the cell surface and mitochondrial damage; however, no increase of abnormalities in the ultrastructure of cilia on living cells was seen. These results support the hypothesis that HNE and PE contribute to the delayed mucociliary clearance and epithelial damage that is observed in patients with chronic bronchial infection.

Full text not available from this repository.

More information

Published date: January 1991
Keywords: Bronchi, Cilia, Endopeptidases, Epithelium, Humans, Microscopy, Electron, Nasal Mucosa, Neutrophils, Pancreatic Elastase, Pseudomonas aeruginosa, Pyocyanine, alpha 1-Antitrypsin, Journal Article, Research Support, Non-U.S. Gov't

Identifiers

Local EPrints ID: 417033
URI: https://eprints.soton.ac.uk/id/eprint/417033
ISSN: 1044-1549
PURE UUID: 74c40875-b3ea-40ee-a986-f8b7c2567c59
ORCID for R. Read: ORCID iD orcid.org/0000-0002-4297-6728

Catalogue record

Date deposited: 17 Jan 2018 17:30
Last modified: 14 Mar 2019 01:36

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