title: IL-17A induces chromatin remodeling promoting IL-8 release in bronchial epithelial cells: Effect of Tiotropium.
abstract: OBJECTIVE
  IL-17A plays a key role in the persistence of airway inflammation, oxidative stress, and reduction of steroid-sensitivity in COPD. We studied the effect of IL-17A on chromatin remodeling and IL-8 production.
  
  
  METHODS
  We measured the levels of IL-8 and IL-17A in induced sputum supernatants (ISS) from healthy controls (HCs), healthy smokers (HSs), and COPD patients by enzyme-linked immunosorbent assay (ELISA). A human bronchial epithelial cell line (16HBE) was stimulated with ISS from HCs, HSs, or COPD subjects. IL-8 was evaluated in 16HBE by Western blot and real-time polymerase chain reaction (PCR). Histone deacetylase 2 (HDAC2), acetyl histone H3 (Ac-His H3) (k9) and inhibitor kappa kinase alpha (IKKα) levels were evaluated in the nuclear extract by Western blot. Finally, we evaluated the effect of IL-17A depletion in ISS, the silencing of IKKα, and the anti-inflammatory effects of Tiotropium Spiriva® (100nM) on 16HBE.
  
  
  RESULTS
  IL-8 and IL-17A levels were higher in ISS from COPD patients and HSs than from HCs. IL-8 protein and messenger RNA (mRNA) levels were increased in 16HBE stimulated with ISS from COPD patients compared with untreated cells. Furthermore, ISS from COPD patients reduced the nuclear levels of HDAC2 while increasing the activity of both Ac-His H3 (k9) and IKKα in stimulated 16HBE. IL-17A depletion in ISS and the IKKα silencing in 16HBE significantly increased the nuclear levels of HDAC2, reduced Ac-His H3 (k9), and promoted IL-8 synthesis in stimulated 16HBE. Tiotropium controls the proinflammatory activity generated by ISS from COPD patients in 16HBE.
  
  
  CONCLUSIONS
  IL-17A present in the airway of COPD patients, which induces chromatin remodeling, promotes the release of IL-8 in the bronchial epithelium. Tiotropium is able to control this proinflammatory activity.
authors.affiliation: Unit: "Ex vivo/In vitro models to study the Immunopathology and Pharmacology of airway diseases", Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR), Palermo, Italy; Dipartimento di Biomedicina sperimentale e Neuroscienze Cliniche (BioNec), University of Palermo, Palermo, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Anzalone
authors.affiliation: Unit: "Ex vivo/In vitro models to study the Immunopathology and Pharmacology of airway diseases", Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Rosalia
authors.initials: R
authors.lastname: Gagliardo
authors.affiliation: Dipartimento di Biomedicina sperimentale e Neuroscienze Cliniche (BioNec), University of Palermo, Palermo, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Bucchieri
authors.affiliation: Unit: "Ex vivo/In vitro models to study the Immunopathology and Pharmacology of airway diseases", Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Giusy Daniela
authors.initials: GD
authors.lastname: Albano
authors.affiliation: Unit: "Ex vivo/In vitro models to study the Immunopathology and Pharmacology of airway diseases", Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Liboria
authors.initials: L
authors.lastname: Siena
authors.affiliation: Unit: "Ex vivo/In vitro models to study the Immunopathology and Pharmacology of airway diseases", Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Angela Marina
authors.initials: AM
authors.lastname: Montalbano
authors.affiliation: Unit: "Ex vivo/In vitro models to study the Immunopathology and Pharmacology of airway diseases", Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Bonanno
authors.affiliation: Unit: "Ex vivo/In vitro models to study the Immunopathology and Pharmacology of airway diseases", Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Riccobono
authors.affiliation: Pharma GmbH & Co. KG, Boehringer Ingelheim, Biberach, Germany.
authors.forename: Michael Paul
authors.initials: MP
authors.lastname: Pieper
authors.affiliation: Unit: "Ex vivo/In vitro models to study the Immunopathology and Pharmacology of airway diseases", Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Mark
authors.initials: M
authors.lastname: Gjomarkaj
authors.affiliation: Unit: "Ex vivo/In vitro models to study the Immunopathology and Pharmacology of airway diseases", Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR), Palermo, Italy. Electronic address: profita@ibim.cnr.it.
authors.forename: Mirella
authors.initials: M
authors.lastname: Profita
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D001980:Bronchi / Q000166:cytology / Q000187:drug effects; D002478:Cells, Cultured; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D056464:Histone Deacetylase 2 / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D020381:Interleukin-17 / Q000378:metabolism*; D016209:Interleukin-8 / Q000378:metabolism*; D029424:Pulmonary Disease, Chronic Obstructive / Q000378:metabolism; D012907:Smoking / Q000378:metabolism; D013183:Sputum / Q000378:metabolism; D000069447:Tiotropium Bromide / Q000494:pharmacology*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; C508606:CXCL8 protein, human; D006657:Histones; C508717:IL17A protein, human; D020381:Interleukin-17; D016209:Interleukin-8; D056464:Histone Deacetylase 2; D000069447:Tiotropium Bromide
keywords: Bronchial epithelial cells; Chromatin remodeling; Chronic obstructive pulmonary disease; IL-17A; IL-8