title: Cigarette smoke combined with Toll-like receptor 3 signaling triggers exaggerated epithelial regulated upon activation, normal T-cell expressed and secreted/CCL5 expression in chronic rhinosinusitis.
abstract: BACKGROUND
  Chronic rhinosinusitis (CRS) is characterized by persistent mucosal inflammation and frequent exacerbations.
  
  
  OBJECTIVE
  To determine whether innate epithelial responses to cigarette smoke or bacterial or viral pathogens may be abnormal in CRS leading to an inappropriate inflammatory response.
  
  
  METHODS
  Primary nasal epithelial cells (PNECs) were grown from middle turbinate biopsies of 9 healthy controls and 11 patients with CRS. After reaching 80% to 90% confluence, PNECs were exposed to medium or cigarette smoke extract (CSE) 5% (vol/vol) for 1 hour, washed, then stimulated with staphylococcal lipoteichoic acid, LPS, or double-stranded RNA (dsRNA). After 24 hours, gene expression was quantified by QRT-PCR.
  
  
  RESULTS
  At baseline, PNECs revealed elevated TNF-alpha and growth-related oncogene-alpha (a C-X-C chemokine)/CXCL1 (GRO-alpha) (4-fold increase, P = .02; and 16-fold increase, P = .004, respectively) in subjects with CRS compared with controls with normal levels of IL-1beta, IL-6, IL-8/CXCL8, human beta-defensin-2, monocyte chemoattractant protein 2/CCL8, monocyte chemoattractant protein 3/CCL7, and regulated upon activation, normal T-cell expressed and secreted (RANTES)/CCL5. Immunostaining of nasal biopsies, however, revealed comparable epithelial staining for TNF-alpha, GRO-alpha, and RANTES. There were no differences in mRNA induction by CSE, TNF-alpha, lipoteichoic acid, LPS, or dsRNA alone. The combination of CSE+dsRNA induced exaggerated RANTES (12,115-fold vs 1500-fold; P = .03) and human beta-defensin-2 (1120-fold vs 12.5-fold; P = .05) in subjects with CRS. No other genes were differentially induced. Furthermore, CSE+dsRNA induced normal levels of IFN-beta, IFN-lambda1, and IFN-lambda2/3 mRNA in subjects with CRS.
  
  
  CONCLUSIONS
  Cigarette smoke extract plus dsRNA induces exaggerated epithelial RANTES expression in patients with CRS. We propose that an analogous response to cigarette smoke plus viral infection may contribute to acute exacerbations and eosinophilic mucosal inflammation in CRS.
authors.affiliation: Division of Rheumatology, Allergy and Immunology, Massachusetts General Hospital, Boston, MA 02114, USA.
authors.forename: Moshe
authors.initials: M
authors.lastname: Yamin
authors.forename: Eric H
authors.initials: EH
authors.lastname: Holbrook
authors.forename: Stacey T
authors.initials: ST
authors.lastname: Gray
authors.forename: Rachel
authors.initials: R
authors.lastname: Harold
authors.forename: Nicolas
authors.initials: N
authors.lastname: Busaba
authors.forename: Avinash
authors.initials: A
authors.lastname: Sridhar
authors.forename: Katia J
authors.initials: KJ
authors.lastname: Powell
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Hamilos
mesh_terms: D000328:Adult; D000368:Aged; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000096:biosynthesis*; D054410:Chemokine CCL7 / Q000096:biosynthesis; D002908:Chronic Disease; D045424:Complex Mixtures / Q000494:pharmacology*; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007120:Immunochemistry; D016899:Interferon-beta / Q000096:biosynthesis; D007372:Interferons; D053583:Interleukin-1beta / Q000096:biosynthesis; D015850:Interleukin-6 / Q000096:biosynthesis; D016209:Interleukin-8 / Q000096:biosynthesis; D007378:Interleukins / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D009297:Nasal Mucosa / Q000378:metabolism* / Q000473:pathology; D012330:RNA, Double-Stranded / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D012220:Rhinitis; D012852:Sinusitis; D014028:Tobacco Smoke Pollution / Q000009:adverse effects*; D051196:Toll-Like Receptor 3 / Q000819:agonists* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D023083:beta-Defensins / Q000096:biosynthesis; D000095022:Interferon Lambda
chemical_list: D018946:Chemokine CCL5; D054410:Chemokine CCL7; D045424:Complex Mixtures; D016899:Interferon-beta; D007372:Interferons; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D007378:Interleukins; D008070:Lipopolysaccharides; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D014028:Tobacco Smoke Pollution; D051196:Toll-Like Receptor 3; D014409:Tumor Necrosis Factor-alpha; D023083:beta-Defensins; C483024:CCL5 protein, human; C515450:CCL7 protein, human; C508606:CXCL8 protein, human; C000624353:interferon-lambda, human; D000095022:Interferon Lambda; C508600:IL6 protein, human; C495344:TLR3 protein, human