title: Neutrophil activation in severe, early-onset COPD patients versus healthy non-smoker subjects in vitro: effects of antioxidant therapy.
abstract: BACKGROUND
  Neutrophils and oxidative stress have been implicated in the pathogenesis of COPD. Severe, early-onset COPD is characterized by a rapid decline in the lung function at an early age; however, nothing is known about neutrophil activation in COPD patients.
  
  
  OBJECTIVE
  The aim of this study was to evaluate peripheral blood neutrophil activation in severe, early-onset COPD patients versus healthy non-smokers and the effect of N-acetyl-L-cysteine (NAC) on neutrophil activation in vitro.
  
  
  METHODS
  Neutrophils were isolated from 15 severe, early-onset COPD patients and 15 age-matched healthy subjects and stimulated with N-formyl-Met-Leu-Phe (fMLP) in the presence or absence of NAC (10 μM to 10 mM). Neutrophil chemotaxis, elastase release, reactive oxygen species (ROS), intracellular thiols and apoptosis were measured by Boyden chamber, spectrofluorometry, CMFDA and H2DCF-DA dyes and by annexin V-FITC, respectively.
  
  
  RESULTS
  Chemotaxis of peripheral blood neutrophils from COPD patients in response to fMLP was 30% more increased than that observed in healthy subjects. Elastase release in response to fMLP was 2-fold higher in neutrophils from COPD patients versus healthy subjects. Intracellular thiol levels were 30% lower in COPD and ROS was approximately 30% higher in COPD versus healthy neutrophils. Spontaneous apoptosis showed no differences in both groups of patients and fMLP-induced apoptosis was higher in COPD. Pre-treatment with the antioxidant NAC effectively attenuated neutrophil chemotaxis, elastase release and ROS as well as effectively increased thiol levels in COPD.
  
  
  CONCLUSIONS
  Neutrophils in severe, early-onset COPD patients are highly activated and this is alleviated by NAC in vitro.
authors.affiliation: Research Unit, University General Hospital Consortium, and Department of Medicine, Faculty of Medicine, University of Valencia, Valencia, Spain. xmilara@hotmail.com
authors.forename: Javier
authors.initials: J
authors.lastname: Milara
authors.forename: Gustavo
authors.initials: G
authors.lastname: Juan
authors.forename: Teresa
authors.initials: T
authors.lastname: Peiró
authors.forename: Adela
authors.initials: A
authors.lastname: Serrano
authors.forename: Julio
authors.initials: J
authors.lastname: Cortijo
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D016022:Case-Control Studies; D002633:Chemotaxis / Q000187:drug effects; D005260:Female; D016166:Free Radical Scavengers / Q000494:pharmacology*; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D019272:Leukocyte Elastase / Q000187:drug effects; D008297:Male; D008875:Middle Aged; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D018375:Neutrophil Activation / Q000187:drug effects*; D009504:Neutrophils / Q000187:drug effects*; D029424:Pulmonary Disease, Chronic Obstructive / Q000503:physiopathology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D016166:Free Radical Scavengers; D016209:Interleukin-8; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019272:Leukocyte Elastase; D000111:Acetylcysteine