Nitric oxide production by polymorphonuclear leucocytes in infected cystic fibrosis sputum consumes oxygen

Kolpen, M; Bjarnsholt, T; Moser, C; Hansen, CR; Rickelt, LF; Kühl, M; Hempel, C; Pressler, T; Høiby, N; Jensen, PO

Nitric oxide production by polymorphonuclear leucocytes in infected cystic fibrosis sputum consumes oxygen

Kolpen, M Bjarnsholt, T Moser, C Hansen, CR Rickelt, LF Kühl, M

Hempel, C Pressler, T Høiby, N Jensen, PO

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Publication Type:: Journal Article
Citation:: Clinical and Experimental Immunology, 2014, 177 (1), pp. 310 - 319
Issue Date:: 2014-07-01

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Field	Value	Language
dc.contributor.author	Kolpen, M	en_US
dc.contributor.author	Bjarnsholt, T	en_US
dc.contributor.author	Moser, C	en_US
dc.contributor.author	Hansen, CR	en_US
dc.contributor.author	Rickelt, LF	en_US
dc.contributor.author	Kühl, M https://orcid.org/0000-0002-1792-4790	en_US
dc.contributor.author	Hempel, C	en_US
dc.contributor.author	Pressler, T	en_US
dc.contributor.author	Høiby, N	en_US
dc.contributor.author	Jensen, PO	en_US
dc.date.available	2014-03-03	en_US
dc.date.issued	2014-07-01	en_US
dc.identifier.citation	Clinical and Experimental Immunology, 2014, 177 (1), pp. 310 - 319	en_US
dc.identifier.issn	0009-9104	en_US
dc.identifier.uri	http://hdl.handle.net/10453/35913
dc.description.abstract	Chronic Pseudomonas aeruginosa lung infection in cystic fibrosis (CF) patients is characterized by persisting mucoid biofilms in hypoxic endobronchial mucus. These biofilms are surrounded by numerous polymorphonuclear leucocytes (PMNs), which consume a major part of present molecular oxygen (O2) due to production of superoxide (O2-). In this study, we show that the PMNs also consume O2 for production of nitric oxide (NO) by the nitric oxide synthases (NOS) in the infected endobronchial mucus. Fresh expectorated sputum samples (n=28) from chronically infected CF patients (n=22) were analysed by quantifying and visualizing the NO production. NO production was detected by optode measurements combined with fluorescence microscopy, flow cytometry and spectrophotometry. Inhibition of nitric oxide synthases (NOS) with NG-monomethyl-L-arginine (L-NMMA) resulted in reduced O2 consumption (P<0·0008, n=8) and a lower fraction of cells with fluorescence from the NO-indicator 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM) (P<0·002, n=8). PMNs stained with DAF-FM and the superoxide indicator hydroethidine (HE) and host cells with inducible NOS (iNOS) were identified in the sputum. In addition, the production of the stable end-products of NO in CF sputum was correlated with the concentration of PMNs; NO3- (P<0·04, r=0·66, n=10) and NO2- (P< 0·006, r=0·78, n=11). The present study suggests that besides consumption of O2 for production of reactive oxygen species, the PMNs in CF sputum also consume O2 for production of NO. © 2014 British Society for Immunology.	en_US
dc.relation.ispartof	Clinical and Experimental Immunology	en_US
dc.relation.isbasedon	10.1111/cei.12318	en_US
dc.subject.classification	Immunology	en_US
dc.subject.mesh	Lung	en_US
dc.subject.mesh	Respiratory Mucosa	en_US
dc.subject.mesh	Neutrophils	en_US
dc.subject.mesh	Cells, Cultured	en_US
dc.subject.mesh	Sputum	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Pseudomonas aeruginosa	en_US
dc.subject.mesh	Pseudomonas Infections	en_US
dc.subject.mesh	Cystic Fibrosis	en_US
dc.subject.mesh	Chronic Disease	en_US
dc.subject.mesh	Nitric Oxide	en_US
dc.subject.mesh	omega-N-Methylarginine	en_US
dc.subject.mesh	Oxygen Consumption	en_US
dc.subject.mesh	Adult	en_US
dc.subject.mesh	Middle Aged	en_US
dc.subject.mesh	Male	en_US
dc.subject.mesh	Nitric Oxide Synthase	en_US
dc.subject.mesh	Young Adult	en_US
dc.title	Nitric oxide production by polymorphonuclear leucocytes in infected cystic fibrosis sputum consumes oxygen	en_US
dc.type	Journal Article
utslib.citation.volume	1	en_US
utslib.citation.volume	177	en_US
utslib.for	1107 Immunology	en_US
utslib.for	0605 Microbiology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - C3 - Climate Change Cluster
utslib.copyright.status	closed_access
pubs.issue	1	en_US
pubs.publication-status	Published	en_US
pubs.volume	177	en_US

Abstract:

Chronic Pseudomonas aeruginosa lung infection in cystic fibrosis (CF) patients is characterized by persisting mucoid biofilms in hypoxic endobronchial mucus. These biofilms are surrounded by numerous polymorphonuclear leucocytes (PMNs), which consume a major part of present molecular oxygen (O2) due to production of superoxide (O2-). In this study, we show that the PMNs also consume O2 for production of nitric oxide (NO) by the nitric oxide synthases (NOS) in the infected endobronchial mucus. Fresh expectorated sputum samples (n=28) from chronically infected CF patients (n=22) were analysed by quantifying and visualizing the NO production. NO production was detected by optode measurements combined with fluorescence microscopy, flow cytometry and spectrophotometry. Inhibition of nitric oxide synthases (NOS) with NG-monomethyl-L-arginine (L-NMMA) resulted in reduced O2 consumption (P<0·0008, n=8) and a lower fraction of cells with fluorescence from the NO-indicator 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM) (P<0·002, n=8). PMNs stained with DAF-FM and the superoxide indicator hydroethidine (HE) and host cells with inducible NOS (iNOS) were identified in the sputum. In addition, the production of the stable end-products of NO in CF sputum was correlated with the concentration of PMNs; NO3- (P<0·04, r=0·66, n=10) and NO2- (P< 0·006, r=0·78, n=11). The present study suggests that besides consumption of O2 for production of reactive oxygen species, the PMNs in CF sputum also consume O2 for production of NO. © 2014 British Society for Immunology.

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