title: Mechanism of interleukin-13 production by granulocyte-macrophage colony-stimulating factor-dependent macrophages via protease-activated receptor-2.
abstract: BACKGROUND
  Granulocyte-macrophage colony-stimulating factor (GM-CSF) promotes classically activated M1 macrophages. GM-CSF upregulates protease-activated receptor-2 (PAR-2) protein expression and activation of PAR-2 by human neutrophil elastase (HNE) regulates cytokine production.
  
  
  OBJECTIVE
  This study investigated the mechanism of PAR-2-mediated interleukin (IL)-13 production by GM-CSF-dependent macrophages stimulated with HNE.
  
  
  METHODS
  Adherent macrophages were obtained from primary cultures of human mononuclear cells. After stimulation with HNE to activate the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling pathway, IL-13 mRNA and protein levels were assessed by the reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay, respectively.
  
  
  RESULTS
  PAR-2 protein was detected in GM-CSF-dependent macrophages by Western blotting. Unexpectedly, PD98059 (an ERK1 inhibitor) increased IL-13 production, even at higher concentrations. Interestingly, U0126 (an ERK1/2 inhibitor) reduced IL-13 production in a concentration-dependent manner. Neither SB203580 (a p38alpha/p38beta inhibitor) nor BIRB796 (a p38gamma/p38delta inhibitor) affected IL-13 production, while TMB-8 (a calcium chelator) diminished IL-13 production.
  
  
  CONCLUSIONS
  Stimulation with HNE promoted the production of IL-13 (a Th2 cytokine) by GM-CSF-dependent M1 macrophages. PAR-2-mediated IL-13 production may be dependent on the Ca(2+)/ERK2 signaling pathway.
authors.affiliation: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan; Graduate School of Medical Science, Kumamoto University Medical SchooL, Kumamoto, Japan.
authors.forename: Rui
authors.initials: R
authors.lastname: Yamaguchi
authors.affiliation: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Takatoshi
authors.initials: T
authors.lastname: Yamamoto
authors.affiliation: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Arisa
authors.initials: A
authors.lastname: Sakamoto
authors.affiliation: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Yasuji
authors.initials: Y
authors.lastname: Ishimaru
authors.affiliation: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Narahara
authors.affiliation: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sugiuchi
authors.affiliation: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Hirose
authors.affiliation: Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan. Electronic address: yamaguti@kumamoto-hsu.ac.jp.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Yamaguchi
mesh_terms: D002070:Butadienes / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005419:Flavonoids / Q000494:pharmacology; D005707:Gallic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D005786:Gene Expression Regulation; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D018793:Interleukin-13 / Q000235:genetics* / Q000276:immunology; D019272:Leukocyte Elastase / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology*; D048052:Mitogen-Activated Protein Kinase 3 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D009281:Naphthalenes / Q000494:pharmacology; D009570:Nitriles / Q000494:pharmacology; D061251:Primary Cell Culture; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D044464:Receptor, PAR-2 / Q000235:genetics* / Q000276:immunology; D015398:Signal Transduction; D048051:p38 Mitogen-Activated Protein Kinases; D010671:Phenylurea Compounds
chemical_list: D002070:Butadienes; D002118:Calcium; D002121:Calcium Channel Blockers; D005419:Flavonoids; D005707:Gallic Acid; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D007093:Imidazoles; D018793:Interleukin-13; D019272:Leukocyte Elastase; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D009281:Naphthalenes; D009570:Nitriles; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011725:Pyridines; D044464:Receptor, PAR-2; D048051:p38 Mitogen-Activated Protein Kinases; C578095:IL13 protein, human; C113580:U 0126; C006014:8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate; C535150:MAPK1 protein, human; C452139:doramapimod; D010671:Phenylurea Compounds; C093642:SB 203580; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
keywords: Extracellular signal-regulated kinase; Granulocyte-macrophage colony-stimulating factor; Interleukin-13; Neutrophil elastase; Protease-activated receptor-2