The effects of leukocytes, local hormones and selected peptides on the permeability of the bronchial mucosa to albumin in vitro
The effects of leukocytes, local hormones and selected peptides on the permeability of the bronchial mucosa to albumin in vitro
An in vitro model system for measuring the integrity of bovine airways mucosa was used to determine the flux of albumin across the airways mucosa. It was shown that stimulated eosinophils and neutrophils added to either the apical or basolateral side were capable of producing an increase in the flux of albumin across the bovine bronchial mucosa. However, no effect was observed in the tracheal mucosa. This effect was inhibited by α2-macroglobulin but was unaffected by heparin, AA-861 (an antioxidant 5-lipoxygenase inhibitor) or cytochalasin b. Further studies using gelatin substrate SDS-PAGE indicated the presence of gelatinase activity in supernatants from the incubation of eosinophils and the bronchial mucosa. Amongst the gelatinase detected were two proteins of molecular masses of 95 and 67 kDa, corresponding to known type IV collagenases. Further studies investigated the effect of other enzymes, collagenase from C. histolyticum and human mast cell tryptase were shown to increase the flux of albumin across the bovine bronchial mucosa. In addition, spent growth medium from house dust mites grown in culture and Der p I, a purified extract from house dust mites were shown to produce a similar effect after activation of the cysteine proteinase activity with DTT. Investigations were continued further by determining the effects of local hormones and selected peptides on the flux of albumin. These produced little effect, contrasting with their reported ability to increase vascular endothelial cell permeability. In conclusion, the results of these studies indicate that activated inflammatory cells, such as eosinophils and neutrophils are capable of producing epithelial damage and increasing the flux of BSA across the bronchial mucosa and that a role for a proteinase is implicated in this process. In addition, the effect of other proteinases indicated the possible importance of such enzymes in the pathology of diseases of the airways.
University of Southampton
Herbert, Carolyn Ann
38b7a402-4cbf-427c-84f6-8059c963e66e
1992
Herbert, Carolyn Ann
38b7a402-4cbf-427c-84f6-8059c963e66e
Herbert, Carolyn Ann
(1992)
The effects of leukocytes, local hormones and selected peptides on the permeability of the bronchial mucosa to albumin in vitro.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
An in vitro model system for measuring the integrity of bovine airways mucosa was used to determine the flux of albumin across the airways mucosa. It was shown that stimulated eosinophils and neutrophils added to either the apical or basolateral side were capable of producing an increase in the flux of albumin across the bovine bronchial mucosa. However, no effect was observed in the tracheal mucosa. This effect was inhibited by α2-macroglobulin but was unaffected by heparin, AA-861 (an antioxidant 5-lipoxygenase inhibitor) or cytochalasin b. Further studies using gelatin substrate SDS-PAGE indicated the presence of gelatinase activity in supernatants from the incubation of eosinophils and the bronchial mucosa. Amongst the gelatinase detected were two proteins of molecular masses of 95 and 67 kDa, corresponding to known type IV collagenases. Further studies investigated the effect of other enzymes, collagenase from C. histolyticum and human mast cell tryptase were shown to increase the flux of albumin across the bovine bronchial mucosa. In addition, spent growth medium from house dust mites grown in culture and Der p I, a purified extract from house dust mites were shown to produce a similar effect after activation of the cysteine proteinase activity with DTT. Investigations were continued further by determining the effects of local hormones and selected peptides on the flux of albumin. These produced little effect, contrasting with their reported ability to increase vascular endothelial cell permeability. In conclusion, the results of these studies indicate that activated inflammatory cells, such as eosinophils and neutrophils are capable of producing epithelial damage and increasing the flux of BSA across the bronchial mucosa and that a role for a proteinase is implicated in this process. In addition, the effect of other proteinases indicated the possible importance of such enzymes in the pathology of diseases of the airways.
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Published date: 1992
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Local EPrints ID: 461270
URI: http://eprints.soton.ac.uk/id/eprint/461270
PURE UUID: a273e603-3a8c-4682-abae-e5649b7e64d1
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Date deposited: 04 Jul 2022 18:42
Last modified: 23 Jul 2022 01:08
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Author:
Carolyn Ann Herbert
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