Nitric oxide production by polymorphonuclear leukocytes in sputum from cystic fibrosis patients with chronic lung infection

TY - ABST

T1 - Nitric oxide production by polymorphonuclear leukocytes in sputum from cystic fibrosis patients with chronic lung infection

AU - Kolpen, Mette

AU - Bjarnsholt, Thomas

AU - Moser, Claus

AU - Hansen, Christine Rønne

AU - Kühl, Michael

AU - Høiby, Niels

AU - Jensen, Peter Østrup

N1 - Conference code: 33

PY - 2010

Y1 - 2010

N2 - Objective: Chronic Pseudomonas aeruginosa lung infection in CF patients is characterized by persisting mucoid biofilm in hypoxic endobronchial mucus. These biofilms are surrounded by numerous polymorphonuclear leukocytes (PMNs), which are the major consumers of O2 for production of O2-. In this study, we hypothesized that nitric oxide (NO) is also produced by the NO synthases (NOS) in the O2 consuming PMNs in the endobronchial mucus.Methods and results: This hypothesis was tested on fresh expectorated sputum from chronically infected CF patients by estimating and visualizing the NO production. A possitive correlation was observed between active PMNs and stable endproducts of NO production; NO3- and NO2- (P< 0.04, r2: 0.66, n = 10, P< 0.006, r2: 0.78, n = 11) using the Griess Reagent System, indicating a production of NO by PMNs. In addition, microsensor analysis revealed the presence of discrete occurrences of NO. By fluorescence microscopy PMNs stained with the NO-indicator, DAF-FM, could be identified. To further verify NO production by the PMNs, active NOS in CF sputum is presently beeing substantiated by estimating O2 consumption and DAF-FM fluorescence during inhibition of NOS.Conclusions: The present study suggests that besides consumption of O2 for O2- production, the PMNs in CF sputum can also consume O2 for NO production. Therefore in addition to consume O2 for the production of reactive oxygen species, the PMNs in CF sputum are also capable of consuming O2 for the generation of reactive nitrogen species.

AB - Objective: Chronic Pseudomonas aeruginosa lung infection in CF patients is characterized by persisting mucoid biofilm in hypoxic endobronchial mucus. These biofilms are surrounded by numerous polymorphonuclear leukocytes (PMNs), which are the major consumers of O2 for production of O2-. In this study, we hypothesized that nitric oxide (NO) is also produced by the NO synthases (NOS) in the O2 consuming PMNs in the endobronchial mucus.Methods and results: This hypothesis was tested on fresh expectorated sputum from chronically infected CF patients by estimating and visualizing the NO production. A possitive correlation was observed between active PMNs and stable endproducts of NO production; NO3- and NO2- (P< 0.04, r2: 0.66, n = 10, P< 0.006, r2: 0.78, n = 11) using the Griess Reagent System, indicating a production of NO by PMNs. In addition, microsensor analysis revealed the presence of discrete occurrences of NO. By fluorescence microscopy PMNs stained with the NO-indicator, DAF-FM, could be identified. To further verify NO production by the PMNs, active NOS in CF sputum is presently beeing substantiated by estimating O2 consumption and DAF-FM fluorescence during inhibition of NOS.Conclusions: The present study suggests that besides consumption of O2 for O2- production, the PMNs in CF sputum can also consume O2 for NO production. Therefore in addition to consume O2 for the production of reactive oxygen species, the PMNs in CF sputum are also capable of consuming O2 for the generation of reactive nitrogen species.

U2 - 10.1016/s1569-1993(10)60188-9

DO - 10.1016/s1569-1993(10)60188-9

M3 - Conference abstract in journal

SN - 1569-1993

SP - S49

JO - Journal of Cystic Fibrosis

JF - Journal of Cystic Fibrosis

IS - Supplement 1

T2 - 33rd European Cystic Fibrosis Conference 2010

Y2 - 19 May 2010 through 16 June 2010

ER -