title: The effects of smoking on the lipopolysaccharide response and glucocorticoid sensitivity of alveolar macrophages of patients with asthma.
abstract: BACKGROUND
  Cigarette smoking in asthma patients causes insensitivity to inhaled glucocorticoids (GCs). We tested the hypothesis that smoking causes GC insensitivity in alveolar macrophages (AMs) obtained from patients with asthma.
  
  
  METHODS
  Nineteen asthmatic nonsmokers (ANSs) and 13 asthmatic smokers (ASMs) underwent BAL. AMs were cultured with or without dexamethasone, 0.1 to 1,000 nmol/L, for 2 h before lipopolysaccharide (LPS) [1 microg/mL] stimulation. After 6 h, supernatants were harvested for enzyme-linked immunosorbent assay, and messenger RNA was collected for real-time (RT)-polymerase chain reaction (PCR).
  
  
  RESULTS
  ASMs had higher numbers of AMs per milliliter of BAL fluid than ANSs (1.98 vs 0.75 x 10(6) cells/mL, respectively; p = 0.007). Cigarette smoking significantly attenuated the LPS response for all three cytokines tested among ANSs vs ASMs (tumor necrosis factor [TNF]-alpha, 31.6 vs 10.6 ng/mL, respectively (p = 0.01); interleukin [IL]-6, 25.8 vs 10.8 ng/mL, respectively (p = 0.002); IL-8, 62.5 vs 36.1 ng/mL, respectively (p = 0.001)). There was no difference in dexamethasone dose-response curves between ANSs and ASMs (p > 0.05 for all comparisons). The inhibitory concentration of 50% (IC(50)) for IL-6 was 120.6 vs 83.3, respectively, and for TNF-alpha it was 4.9 vs 8.6, respectively; an IC(50) was not achieved for IL-8. RT-PCR also showed no difference in the suppression of cytokine messenger RNA levels between groups, with IL-8 being the most GC-insensitive cytokine.
  
  
  CONCLUSIONS
  Cigarette smoking in patients with asthma increases the number of airway AMs and attenuates their response to LPS, which may have implications in host immune function. Cigarette smoking does not alter the GC sensitivity of AMs in patients with asthma. There was differential cytokine sensitivity, with IL-8 being the least GC-sensitive cytokine. GC-insensitive IL-8 production from AMs may be a mechanism by which neutrophils are attracted into the airways.
authors.affiliation: School of Translational Medicine, University of Manchester, University Hospital of South Manchester Foundation Trust, Manchester, UK. Electronic address: binitakane@yahoo.com.
authors.forename: Binita
authors.initials: B
authors.lastname: Kane
authors.affiliation: School of Translational Medicine, University of Manchester, University Hospital of South Manchester Foundation Trust, Manchester, UK.
authors.forename: Umme
authors.initials: U
authors.lastname: Kolsum
authors.affiliation: School of Translational Medicine, University of Manchester, University Hospital of South Manchester Foundation Trust, Manchester, UK.
authors.forename: Thomas
authors.initials: T
authors.lastname: Southworth
authors.affiliation: School of Translational Medicine, University of Manchester, University Hospital of South Manchester Foundation Trust, Manchester, UK.
authors.forename: Jane
authors.initials: J
authors.lastname: Armstrong
authors.affiliation: School of Translational Medicine, University of Manchester, University Hospital of South Manchester Foundation Trust, Manchester, UK.
authors.forename: Ashley
authors.initials: A
authors.lastname: Woodcock
authors.affiliation: School of Translational Medicine, University of Manchester, University Hospital of South Manchester Foundation Trust, Manchester, UK.
authors.forename: Dave
authors.initials: D
authors.lastname: Singh
mesh_terms: D000328:Adult; D001249:Asthma / Q000378:metabolism / Q000473:pathology*; D016022:Case-Control Studies; D018929:Cell Culture Techniques; D003907:Dexamethasone / Q000494:pharmacology*; D005260:Female; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D012907:Smoking / Q000009:adverse effects*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D055815:Young Adult
chemical_list: D005938:Glucocorticoids; D015850:Interleukin-6; D016209:Interleukin-8; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone