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.
Aim:
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.
Discussion:
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.
Keywords:
Extracellular signal-regulated kinase; Granulocyte-macrophage colony-stimulating factor; Interleukin-13; Neutrophil elastase; Protease-activated receptor-2.
Copyright © 2015 Elsevier Inc. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Butadienes / pharmacology
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Calcium / metabolism*
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Calcium Channel Blockers / pharmacology
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Cell Adhesion
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Cell Differentiation / drug effects
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Dose-Response Relationship, Drug
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Flavonoids / pharmacology
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Gallic Acid / analogs & derivatives
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Gallic Acid / pharmacology
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Gene Expression Regulation
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Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology*
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Humans
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Imidazoles / pharmacology
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Interleukin-13 / genetics*
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Interleukin-13 / immunology
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Leukocyte Elastase / pharmacology
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Macrophages / cytology
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Macrophages / drug effects*
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Macrophages / immunology
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Mitogen-Activated Protein Kinase 1 / antagonists & inhibitors
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Mitogen-Activated Protein Kinase 1 / genetics
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Mitogen-Activated Protein Kinase 1 / immunology*
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Mitogen-Activated Protein Kinase 3 / antagonists & inhibitors
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Mitogen-Activated Protein Kinase 3 / genetics
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Mitogen-Activated Protein Kinase 3 / immunology
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Monocytes / cytology
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Monocytes / drug effects
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Monocytes / immunology
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Naphthalenes / pharmacology
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Nitriles / pharmacology
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Primary Cell Culture
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Protein Kinase Inhibitors / pharmacology
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Pyrazoles / pharmacology
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Pyridines / pharmacology
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Receptor, PAR-2 / genetics*
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Receptor, PAR-2 / immunology
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Signal Transduction
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p38 Mitogen-Activated Protein Kinases
Substances
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Butadienes
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Calcium Channel Blockers
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Flavonoids
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IL13 protein, human
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Imidazoles
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Interleukin-13
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Naphthalenes
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Nitriles
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Protein Kinase Inhibitors
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Pyrazoles
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Pyridines
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Receptor, PAR-2
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U 0126
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8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate
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Gallic Acid
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Granulocyte-Macrophage Colony-Stimulating Factor
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MAPK1 protein, human
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Mitogen-Activated Protein Kinase 1
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Mitogen-Activated Protein Kinase 3
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p38 Mitogen-Activated Protein Kinases
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Leukocyte Elastase
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doramapimod
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SB 203580
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2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
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Calcium