Supplementary Materials and Methods 1 from Neutrophil Elastase Remodels Mammary Tumors to Facilitate Lung Metastasis
Amriti R. LullaSaïd AkliCansu KarakaşJoseph A. CarusoLucas D. WarmaNatalie W. FowlkesXiayu RaoJing WangKelly K. HuntStephanie S. WatowichKhandan Keyomarsi
0
Citation
0
Reference
10
Related Paper
Abstract:
<p>Supplementary Materials and Methods, clean copy version</p>Keywords:
Neutrophil elastase
The persistently dominant activity of neutrophil elastase in bronchial secretions replete with antielastases is crucial to the pathogenesis of bronchiectasis. We hypothesize that components in the bronchial secretions bind neutrophil elastase and compromise the inhibitory efficiency of prevailing antielastases. Zymographic analysis of sputum sols from patients with bronchiectasis found elastase activity in a polydisperse, alcian blue-stained zone of high molecular mass. This suggested that neutrophil elastase was complexed with polyanionic partners. Western blot analysis found not only the polyanionic partner, heparan sulfate/syndecan-1, but also the physiological antielastases, secretory leukoproteinase inhibitor and α1-antitrypsin, in the complex. Both dissociative density gradient ultracentrifugation and heparin displacement revealed that elastase dissociated from heparan sulfate/syndecan-1 was fully inhibited by the endogenous antielastases. This contrasts with the effects of exogenous antielastases on sputum neutrophil elastase activity—that of α1-antitrypsin was limited, but that of secretory leukoproteinase inhibitor was facilitated. Similarly, complexed elastase on blots of sputum sol zymographs was bound and inhibited by exogenous secretory leukoproteinase inhibitor but not by exogenous α1-antitrypsin. Taken together, the results bring a new focus to heparan sulfate/syndecan-1 complexed with neutrophil elastase in inflamed bronchial secretions as a target for modulating elastase susceptibility to physiological antielastases.
Neutrophil elastase
Pathogenesis
Pancreatic elastase
Chronic bronchitis
Cite
Citations (57)
To examine the mechanism of tissue damage which causes bronchiolectasis in diffuse pan-bronchiolitis (DPB), the cellular components, elastase and its main inhibitor, alpha 1-protease inhibitor (α1-PI) were measured in bronchoalveolar and bronchial lavage fluid (BALF and BLF) from 14 DPB patients. A predominant increase in the neutrophil count was observed in DPB. Elastase activity in BALF and BLF was about 1, 000-fold higher in the DPB group than in the control group. An inhibitor study and a positive correlation between elastase activity and the neutrophil count in both lavage fluids from the DPB group indicated that the activity was mainly that of neutrophil elastase. Western blot analysis of αl-PI showed that most of the α1-PI in the lavage fluids from DPB group was degraded. These results indicated that neutrophil infiltration increases the level of elastase in the DPB lesions ; this increase seems to be closely related to tissue damage.(Internal Medicine 31 : 599-605, 1992)
Diffuse panbronchiolitis
Neutrophil elastase
Protease inhibitor (pharmacology)
Cite
Citations (10)
BACKGROUND: DNA released by degenerating inflammatory neutrophils contributes to mucous plugging of airways in patients with cystic fibrosis. Neutrophil elastase, a major effector of tissue destruction in the lungs of patients with cystic fibrosis, is a highly cationic molecule which is bound and inhibited by negatively charged polyanions such as mucin and DNA in purulent sputum. Thus, the solubilisation of DNA in the airways by aerosolised recombinant DNase may remove a source of neutrophil elastase inhibition, effectively increasing elastase load. The aim of this study was to assess the effect of rhDNase therapy on neutrophil elastase load in patients with cystic fibrosis. METHODS: Blood and sputum were collected from 15 patients with cystic fibrosis before initiation of nebulised DNase therapy and at 12 weeks following therapy. The long term effects of continuous rhDNase administration were evaluated at 52 weeks for 11 of these patients. Plasma was analysed for neutrophil elastase, interleukin (IL)-8 and neutrophil elastase in complex with alpha 1-protease inhibitor (alpha 1PI). Sputum was assessed for neutrophil elastase, IL-8, and active elastase. At each visit spirometric measurements were carried out. RESULTS: Sputum elastase activity decreased at 12 weeks and was maintained at 52 weeks when a decline in total plasma elastase was also observed. Although, as expected, there was a correlation between plasma levels of total elastase and neutrophil elastase/alpha 1PI complex, the decrease in the levels of the complex at 52 weeks did not reach statistical significance. CONCLUSIONS: This study indicates that prolonged daily administration of rhDNase results in a reduction in elastase load in patients with cystic fibrosis.
Neutrophil elastase
Pancreatic elastase
Cite
Citations (33)
Neutrophil elastase is a serine protease that has been implicated in the pathogenesis of inflammatory bowel disease. Due to post-translational control of its activation and high expression of its inhibitors in the gut, measurements of total expression poorly reflect the pool of active, functional neutrophil elastase. Fluorogenic substrate probes have been used to measure neutrophil elastase activity, though these tools lack specificity and traceability. PK105 is a recently described fluorescent activity-based probe, which binds to neutrophil elastase in an activity-dependent manner. The irreversible nature of this probe allows for accurate identification of its targets in complex protein mixtures. We describe the reactivity profile of PK105b, a new analogue of PK105, against recombinant serine proteases and in tissue extracts from healthy mice and from models of inflammation induced by oral cancer and Legionella pneumophila infection. We apply PK105b to measure neutrophil elastase activation in an acute model of experimental colitis. Neutrophil elastase activity is detected in inflamed, but not healthy, colons. We corroborate this finding in mucosal biopsies from patients with ulcerative colitis. Thus, PK105b facilitates detection of neutrophil elastase activity in tissue lysates, and we have applied it to demonstrate that this protease is unequivocally activated during colitis.
Neutrophil elastase
Inflammatory Bowel Diseases
Cite
Citations (24)
Background - DNA released by degenerating inflammatory neutrophils contributes tomucousplugging ofairways in patients withcystic fibrosis. Neutrophil elastase, a majoreffector oftissue destruction inthelungsofpatients with cysticfibrosis, is a highlycationic moleculewhichisboundandinhibited bynegatively charged polyanions suchas mucinand DNA inpurulent sputum. Thus,thesolubilisation ofDNA inthe airwaysby aerosolised recombinant DNasemayremoveasourceofneutrophil elastase inhibition, effectively increasing elastase load. Theaimofthisstudy wasto assesstheeffect ofrhDNasetherapy on neutrophil elastase loadinpatients with cystic fibrosis. Methods- Bloodandsputumwerecollected from15patients withcystic fibrosis before initiation ofnebulised DNasetherapyandat12weeksfollowing therapy. The longtermeffects ofcontinuous rhDNase administration wereevaluated at52weeks for11ofthesepatients. Plasmawasanalysedforneutrophil elastase, interleukin (IL)-8 andneutrophil elastase incomplex witha,-protease inhibitor (aIPI). Sputum wasassessed forneutrophil elastase, IL-8, andactive elastase. Ateachvisit spirometricmeasurements werecarried out. Results - Sputumelastase activity decreased at12weeksandwasmaintained at52weekswhenadecline intotal plasma elastase wasalsoobserved. Although, as expected, therewasacorrelation between plasmalevels oftotal elastase andneutrophilelastase/alPI complex, thedecrease in thelevels ofthecomplex at52weeksdid notreachstatistical significance. Conclusions - Thisstudyindicates that prolongeddailyadministration of rhDNaseresults inareduction inelastase loadinpatients withcystic fibrosis. (Thorax 1996;51:619-623)
Neutrophil elastase
Pancreatic elastase
Cite
Citations (0)
Mutant tight-skin mice that had severe pulmonary emphysema were crossed with beige mice that contained neutrophils deficient in elastase activity. The resultant tight skin-beige cross mice were deficient in neutrophil elastase activity, yet they were still afflicted with the same degree of severe emphysema as the tight-skin mice. The results indicate that neutrophil elastase does not cause the emphysematous lesions found in tight-skin mice.
Neutrophil elastase
Pulmonary emphysema
Pancreatic elastase
Cite
Citations (19)
Neutrophil elastase
Neutrophil Extracellular Traps
Cite
Citations (18)
Cite
Citations (38)
Neutrophil elastase
Neutrophil Extracellular Traps
Pancreatic elastase
Cite
Citations (18)
Neutrophil-derived elastase promotes destruction of the tissue,increases vascular permeability,and promotes neutrophil migration.Neutrophil elastase inhibitors(NEI) reduce tissue injury which through inhibition of neutrophil elastase(NE) activity.They also have anti-inflammatory and immunomodulatory properties.Recent studies found that NEI provide protection against organ damage in particular of neutrophil-mediated tissue injury.This review was focused on the role of NEI in some diseases.
Neutrophil elastase
Neutrophil Extracellular Traps
Vascular permeability
Cite
Citations (0)