title: Regulation of interleukin-33 and thymic stromal lymphopoietin in human nasal fibroblasts by proinflammatory cytokines.
abstract: OBJECTIVE
  Epithelial-derived interleukin (IL)-33 and thymic stromal lymphopoietin (TSLP) are critical regulators of innate and adaptive immune responses associated with Th2 cytokine-mediated inflammation, including allergic rhinitis. IL-33 and TSLP are expressed not only in epithelial cells but also fibroblasts, endothelial cells, and smooth muscle cells at nasal mucosal sites. However, the role and the regulation of IL-33 and TSLP in nasal fibroblasts remain unknown. We investigated the signal transduction regulation of IL-33 and TSLP induced by proinflammatory cytokines in nasal fibroblasts.
  
  
  METHODS
  In vitro, prospective study.
  
  
  METHODS
  Nasal fibroblasts were derived from human nasal mucosa without allergic rhinitis. Expression of IL-33 and TSLP was examined in nasal fibroblasts treated with proinflammatory cytokines IL-1β or tumor necrosis factor (TNF)-α after pretreatment with or without various inhibitors of signal transduction pathways.
  
  
  RESULTS
  In nasal fibroblasts, both Western blotting and reverse transcriptase-polymerase chain reaction demonstrated that expression of mRNAs and proteins of IL-33 and TSLP was increased by treatment with IL-1β or TNF-α. Immunostaining revealed that IL-33-positive nuclei were markedly increased by the treatment with IL-1β or TNF-α. Enzyme-linked immunosorbent assay showed that fibroblast-released TSLP was significantly increased by treatment with IL-1β or TNF-α. The upregulation of both IL-33 and TSLP proteins by treatment with IL-1β was prevented by inhibitors of pan- protein kinase C (PKC), p38 mitogen-activated protein kinase, and nuclear factor (NF)-κB. In the cells treated with TNF-α, upregulation of IL-33 protein was prevented by inhibitors of phosphoinositide 3-kinase (PI3K), c-Jun N-terminal kinase (JNK), and NF-κB, whereas upregulation of TSLP protein was prevented by inhibitors of pan-PKC, PI3K, JNK, and NF-κB.
  
  
  CONCLUSIONS
  Expression of IL-33 and TSLP in nasal fibroblasts was regulated via distinct signal transduction pathways including NF-κB.
authors.affiliation: Department of Otolaryngology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Nomura
authors.forename: Takashi
authors.initials: T
authors.lastname: Kojima
authors.forename: Jun
authors.initials: J
authors.lastname: Fuchimoto
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Obata
authors.forename: Takashi
authors.initials: T
authors.lastname: Keira
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Himi
authors.forename: Norimasa
authors.initials: N
authors.lastname: Sawada
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D016207:Cytokines / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D053583:Interleukin-1beta / Q000494:pharmacology*; D000067596:Interleukin-33; D007378:Interleukins / Q000378:metabolism*; D009297:Nasal Mucosa / Q000166:cytology*; D012333:RNA, Messenger / Q000032:analysis; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012494:Sampling Studies; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D000094632:Thymic Stromal Lymphopoietin
chemical_list: D016207:Cytokines; C506536:IL33 protein, human; D053583:Interleukin-1beta; D000067596:Interleukin-33; D007378:Interleukins; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D000094632:Thymic Stromal Lymphopoietin