title: Viral stimuli trigger exaggerated thymic stromal lymphopoietin expression by chronic obstructive pulmonary disease epithelium: role of endosomal TLR3 and cytosolic RIG-I-like helicases.
abstract: BACKGROUND
  Rhinovirus (RV)-induced chronic obstructive pulmonary disease (COPD) exacerbations exhibit TH(2)-like inflammation. We hypothesized that RV-infected bronchial epithelial cells (BEC) overproduce TH(2)-switching hub cytokine, thymic stromal lymphopoietin (TSLP) in COPD.
  
  
  METHODS
  Primary BEC from healthy (HBEC) and from COPD donors (COPD-BEC) were grown in 12-well plates, infected with RV16 (0.5-5 MOI) or stimulated with agonists for either toll-like receptor (TLR) 3 (dsRNA, 0.1-10 μg/ml) or RIG-I-like helicases (dsRNA-LyoVec, 0.1-10 μg/ml). Cytokine mRNA and protein were determined (RTqPCR; ELISA).
  
  
  RESULTS
  dsRNA dose-dependently evoked cytokine gene overproduction of TSLP, CXCL8 and TNF-α in COPD-BEC compared to HBEC. This was confirmed using RV16 infection. IFN-β induction did not differ between COPD-BEC and HBEC. Endosomal TLR3 inhibition by chloroquine dose-dependently inhibited dsRNA-induced TSLP generation and reduced generation of CXCL8, TNF-α, and IFN-β. Stimulation of cytosolic viral sensors (RIG-I-like helicases) with dsRNA-LyoVec increased production of CXCL8, TNF-α, and IFN-β, but not TSLP.
  
  
  CONCLUSIONS
  Endosomal TLR3-stimulation, by dsRNA or RV16, induces overproduction of TSLP in COPD-BEC. dsRNA- and RV-induced overproduction of TNF-α and CXCL8 involves endosomal TLR3 and cytosolic RIG-I-like helicases and so does the generation of IFN-β in COPD-BEC. RV16 and dsRNA-induced epithelial TSLP may contribute to pathogenic effects at exacerbations and development of COPD.
authors.affiliation: Unit of Respiratory Immunopharmacology, Lund University Sweden, Lund, Sweden.
authors.forename: Jenny
authors.initials: J
authors.lastname: Calvén
authors.forename: Yulyana
authors.initials: Y
authors.lastname: Yudina
authors.forename: Oskar
authors.initials: O
authors.lastname: Hallgren
authors.forename: Gunilla
authors.initials: G
authors.lastname: Westergren-Thorsson
authors.forename: Donna E
authors.initials: DE
authors.lastname: Davies
authors.forename: Angelica
authors.initials: A
authors.lastname: Brandelius
authors.forename: Lena
authors.initials: L
authors.lastname: Uller
mesh_terms: D000368:Aged; D016207:Cytokines / Q000276:immunology* / Q000378:metabolism; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000276:immunology* / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D010850:Picornaviridae Infections / Q000276:immunology* / Q000378:metabolism; D029424:Pulmonary Disease, Chronic Obstructive / Q000276:immunology* / Q000378:metabolism / Q000473:pathology / Q000821:virology; D012330:RNA, Double-Stranded / Q000276:immunology / Q000494:pharmacology; D011971:Receptors, Immunologic; D020545:Respiratory Mucosa / Q000276:immunology* / Q000473:pathology / Q000821:virology; D012229:Rhinovirus / Q000276:immunology* / Q000378:metabolism; D051196:Toll-Like Receptor 3 / Q000276:immunology* / Q000378:metabolism; D000094632:Thymic Stromal Lymphopoietin
chemical_list: D016207:Cytokines; D012330:RNA, Double-Stranded; D011971:Receptors, Immunologic; C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; C452820:RIGI protein, human; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases; D000094632:Thymic Stromal Lymphopoietin