title: Toll-like receptor-4 mediates cigarette smoke-induced cytokine production by human macrophages.
abstract: BACKGROUND
  The major risk factor for the development of COPD is cigarette smoking. Smoking causes activation of resident cells and the recruitment of inflammatory cells into the lungs, which leads to release of pro-inflammatory cytokines, chemotactic factors, oxygen radicals and proteases. In the present study evidence is found for a new cellular mechanism that refers to a link between smoking and inflammation in lungs.
  
  
  METHODS
  Employing human monocyte-derived macrophages, different techniques including FACS analysis, Cytometric Bead Array Assay and ELISA were achieved to evaluate the effects of CS on pro-inflammatory cytokine secretion including IL-8. Then, Toll-like receptor neutralization was performed to study the involvement of Toll-like receptor-4 in IL-8 production. Finally, signaling pathways in macrophages after exposure to CS medium were investigated performing ELISA and Western analysis.
  
  
  RESULTS
  We demonstrate that especially human monocytes are sensitive to produce IL-8 upon cigarette smoke stimulation compared to lymphocytes or neutrophils. Moreover, monocyte-derived macrophages produce high amounts of the cytokine. The IL-8 production is dependent on Toll-like receptor 4 stimulation and LPS is not involved. Further research resolved the cellular mechanism by which cigarette smoke induces cytokine production in monocyte-derived macrophages. Cigarette smoke causes subsequently a concentration-dependent phosphorylation of IRAK and degradation of TRAF6. Moreover, IkappaBalpha was phosphorylated which suggests involvement of NF-kappaB. In addition, NFkappaB-inhibitor blocked cigarette smoke-induced IL-8 production.
  
  
  CONCLUSIONS
  These findings link cigarette smoke to inflammation and lead to new insights/therapeutic strategies in the pathogenesis of lung emphysema.
authors.affiliation: Department of Pharmacology and Pathophysiology, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, PO BOX 80,082, 3508 TB Utrecht, The Netherlands. karimik@mcmaster.ca
authors.forename: Khalil
authors.initials: K
authors.lastname: Karimi
authors.forename: Hadi
authors.initials: H
authors.lastname: Sarir
authors.forename: Esmaeil
authors.initials: E
authors.lastname: Mortaz
authors.forename: Joost J
authors.initials: JJ
authors.lastname: Smit
authors.forename: Hossein
authors.initials: H
authors.lastname: Hosseini
authors.forename: Sjef J
authors.initials: SJ
authors.lastname: De Kimpe
authors.forename: Frans P
authors.initials: FP
authors.lastname: Nijkamp
authors.forename: Gert
authors.initials: G
authors.lastname: Folkerts
mesh_terms: D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis*; D006801:Humans; D053592:Interleukin-1 Receptor-Associated Kinases; D016209:Interleukin-8 / Q000096:biosynthesis; D047908:Intracellular Signaling Peptides and Proteins / Q000502:physiology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D016328:NF-kappa B / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology; D012906:Smoke; D048029:TNF Receptor-Associated Factor 6 / Q000502:physiology; D014026:Nicotiana; D051197:Toll-Like Receptor 4 / Q000502:physiology*
chemical_list: D003470:Culture Media; D016207:Cytokines; D016209:Interleukin-8; D047908:Intracellular Signaling Peptides and Proteins; D016328:NF-kappa B; D012906:Smoke; D048029:TNF Receptor-Associated Factor 6; D051197:Toll-Like Receptor 4; D053592:Interleukin-1 Receptor-Associated Kinases; D017346:Protein Serine-Threonine Kinases