title: Neutrophil-derived matrix metalloproteinase-9 is increased in severe asthma and poorly inhibited by glucocorticoids.
abstract: BACKGROUND
  Matrix metalloproteinase (MMP)-9 levels are increased in bronchoalveolar lavage (BAL) fluid from patients with severe asthma on high doses of glucocorticoids (GCs).
  
  
  OBJECTIVE
  We sought to identify neutrophils as the source of increased BAL fluid MMP-9 in severe asthma and to evaluate the effects of GCs on this MMP-9.
  
  
  METHODS
  MMP-9 protein, activity, and mRNA were measured in BAL fluid and cells at baseline, and after in vitro GCs in patients with severe asthma and controls using enzyme immunoassays, zymography, Western blotting, and real-time PCR.
  
  
  RESULTS
  The high molecular weight (HMW) form of MMP-9 was significantly increased in severe asthma (P =.02). Western blotting confirmed a heterodimer of MMP-9 and neutrophil gelatinase-associated lipocalin. The HMW MMP-9 correlated with BAL neutrophils (r =.65, P <.0001). BAL cell supernatant MMP-9 protein levels also tended to be higher in patients with severe asthma (overall, P =.09), whereas the HMW activity form was increased (P =.03). MMP-9 protein (and HMW activity) correlated with neutrophils in the cell pellet (r =.75, P <.0001). In contrast to protein and activity, BAL cell mRNA levels were marginally lower in patients with severe asthma than in control subjects (overall, P =.06). Although GCs decreased BAL cell MMP-9 protein and mRNA in vitro, the effect was significantly smaller in severe asthma (P <.01 for both). GCs decreased the pro-MMP-9 activity in patients with severe asthma and normal control subjects, while having no effects on the HMW form (P =.22). Peripheral blood neutrophil MMP-9 protein was not affected by GCs.
  
  
  CONCLUSIONS
  BAL neutrophils contribute to BAL fluid MMP-9 protein and activity and are poorly inhibited by GCs.
authors.affiliation: Department of Medicine, National Jewish Center and Research Center, University of Colorado Health Sciences Center, Denver, CO 80206, USA.
authors.forename: Meghan
authors.initials: M
authors.lastname: Cundall
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Sun
authors.forename: Christina
authors.initials: C
authors.lastname: Miranda
authors.forename: John B
authors.initials: JB
authors.lastname: Trudeau
authors.forename: Stephen
authors.initials: S
authors.lastname: Barnes
authors.forename: Sally E
authors.initials: SE
authors.lastname: Wenzel
mesh_terms: D000209:Acute-Phase Proteins / Q000378:metabolism; D000328:Adult; D001249:Asthma / Q000097:blood / Q000201:enzymology*; D015153:Blotting, Western; D018893:Bronchoalveolar Lavage; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D000071068:Lipocalin-2; D054834:Lipocalins; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism*; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology*; D015513:Oncogene Proteins / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D000209:Acute-Phase Proteins; D005938:Glucocorticoids; C489506:LCN2 protein, human; D000071068:Lipocalin-2; D054834:Lipocalins; D015513:Oncogene Proteins; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D003907:Dexamethasone; D020780:Matrix Metalloproteinase 9