title: Proinflammatory mediators alter expression of nuclear factor kappa B-regulating deubiquitinases in sinonasal epithelial cells.
abstract: BACKGROUND
  Nuclear factor κB (NF-κB) is a vital transcription factor that is activated by numerous inflammatory stimuli. Its activity is tightly regulated by a family of deubiquitinating enzymes (A20, Cezanne, cylindromatosis [CYLD]) that function in a negative-feedback loop, a process that prevents chronic and systemic inflammation. This study seeks to characterize the expression and functional role of NF-κB-regulating deubiquitinases in the sinonasal epithelium.
  
  
  METHODS
  Expression of A20, Cezanne, and CYLD was assessed in normal sinonasal tissue using immunohistochemistry. Cultured sinonasal epithelial cells (SNECs) were stimulated with proinflammatory cytokines (tumor necrosis factor α [TNF-α], interleukin 4 [IL]-4, IL-13) or lipopolysaccharide (LPS) and changes in NF-κB activation and deubiquitinase expression were assessed using Western blots and quantitative real-time polymerase chain reaction (qRT-PCR), respectively.
  
  
  RESULTS
  NF-κB was activated in response to LPS and TNF-α, but not IL-4 or IL-13. A20, Cezanne, and CYLD were all expressed in sinonasal tissue, primarily along the apical surface of the epithelium. Proinflammatory mediators primarily affected expression of A20, with upregulation by LPS and TNF-α and downregulation by IL-4 and IL-13.
  
  
  CONCLUSIONS
  The NF-κB-regulating deubiquitinases A20, Cezanne, and CYLD are expressed in sinonasal tissue and are differentially induced by proinflammatory cytokines and the microbial antigen, LPS. These results suggest an important role for NF-κB-regulating deubiquitinases in mucosal immunity and homeostasis.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Vanderbilt University School of Medicine, Nashville, TN.
authors.forename: Ping
authors.initials: P
authors.lastname: Li
authors.affiliation: Department of Rhinology, The First Affiliated Hospital of ZhengZhou University, ZhengZhou, Henan, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Vanderbilt University School of Medicine, Nashville, TN.
authors.forename: Justin H
authors.initials: JH
authors.lastname: Turner
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000378:metabolism; D000074749:Deubiquitinating Enzyme CYLD; D010450:Endopeptidases / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000517:prevention & control; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D016328:NF-kappa B / Q000378:metabolism*; D009297:Nasal Mucosa / Q000187:drug effects* / Q000201:enzymology; D009687:Nuclear Proteins / Q000378:metabolism; D010256:Paranasal Sinuses / Q000187:drug effects* / Q000201:enzymology; D060888:Real-Time Polymerase Chain Reaction; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D064570:Ubiquitin-Specific Proteases / Q000378:metabolism*
chemical_list: D016207:Cytokines; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides; D016328:NF-kappa B; D009687:Nuclear Proteins; D025521:Tumor Suppressor Proteins; D010450:Endopeptidases; C476458:OTUD7B protein, human; C409984:CYLD protein, human; D000074749:Deubiquitinating Enzyme CYLD; C076529:TNFAIP3 protein, human; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3; D064570:Ubiquitin-Specific Proteases
keywords: A20; CYLD; Cezanne; NF-κB; TNFAIP3; deubiquitinase; epithelial cell; inflammation; rhinosinusitis; sinonasal