Abstract
Background ML-1 (IL-17F) is a recently discovered cytokine, and its function remains elusive. GM-CSF is a crucial cytokine for the maturation of various cell types and regulates allergic airway inflammation. Objective The functional effect of ML-1 in the expression of GM-CSF was investigated. Methods The levels of gene and protein expression in normal human bronchial epithelial cells (NHBEs) in the presence or absence of various kinase inhibitors or, in some cases, of a Raf1 dominant-negative mutant were determined by RT-PCR and ELISA, respectively. Western blotting was performed to investigate kinase activation. Results The results showed first that ML-1 induces, in a time-dependent and dose-dependent manner, the gene and protein expression for GM-CSF NHBEs, which are associated with activation of Raf1 and MAP kinase kinase (MEK) kinases. Selective MEK inhibitors, PD98059 and U0126, and Raf1 kinase inhibitor I significantly inhibited ML-1-induced GM-CSF production. Furthermore, overexpression of Raf1 dominant-negative mutants inhibited IL-17F-induced GMCSF expression. The combination of PD98059 and Raf1 kinase inhibitor I completely blocked GM-CSF production, whereas 2 protein kinase C inhibitors, Ro-31-7549 and GF109203X, and a phosphatidylinositol 3-kinase inhibitor, LY294002, showed no inhibitory effect. Conclusion These findings suggest that ML-1 induces GM-CSF expression through the activation of the Raf1-MEK-extracellular signal-regulated kinase 1/2 pathway.
| Original language | English |
|---|---|
| Pages (from-to) | 444-450 |
| Number of pages | 7 |
| Journal | Journal of Allergy and Clinical Immunology |
| Volume | 114 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Aug 1 2004 |
| Externally published | Yes |
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Keywords
- Bronchial epithelial cells
- Dimethyl sulfoxide
- DMSO
- ERK
- ERK1/2
- Extracellular signal-regulated kinase
- GM-CSF
- MAP kinase
- MAP kinase kinase
- MEK
- NF-κB
- NHBE
- Normal human bronchial epithelial cell
- Nuclear factor κB
- PI3K
- Raf1
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology
Cite this
Induction of granulocyte-macrophage colony-stimulating factor by a new cytokine, ML-1 (IL-17F), via Raf I-MEK-ERK pathway. / Kawaguchi, Mio; Kokubu, Fumio; Odaka, Miho; Watanabe, Shin; Suzuki, Shintaro; Ieki, Koushi; Matsukura, Satoshi; Kurokawa, Masatsugu; Adachi, Mitsuru; Huang, Shau Ku.
In: Journal of Allergy and Clinical Immunology, Vol. 114, No. 2, 01.08.2004, p. 444-450.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Induction of granulocyte-macrophage colony-stimulating factor by a new cytokine, ML-1 (IL-17F), via Raf I-MEK-ERK pathway
AU - Kawaguchi, Mio
AU - Kokubu, Fumio
AU - Odaka, Miho
AU - Watanabe, Shin
AU - Suzuki, Shintaro
AU - Ieki, Koushi
AU - Matsukura, Satoshi
AU - Kurokawa, Masatsugu
AU - Adachi, Mitsuru
AU - Huang, Shau Ku
PY - 2004/8/1
Y1 - 2004/8/1
N2 - Background ML-1 (IL-17F) is a recently discovered cytokine, and its function remains elusive. GM-CSF is a crucial cytokine for the maturation of various cell types and regulates allergic airway inflammation. Objective The functional effect of ML-1 in the expression of GM-CSF was investigated. Methods The levels of gene and protein expression in normal human bronchial epithelial cells (NHBEs) in the presence or absence of various kinase inhibitors or, in some cases, of a Raf1 dominant-negative mutant were determined by RT-PCR and ELISA, respectively. Western blotting was performed to investigate kinase activation. Results The results showed first that ML-1 induces, in a time-dependent and dose-dependent manner, the gene and protein expression for GM-CSF NHBEs, which are associated with activation of Raf1 and MAP kinase kinase (MEK) kinases. Selective MEK inhibitors, PD98059 and U0126, and Raf1 kinase inhibitor I significantly inhibited ML-1-induced GM-CSF production. Furthermore, overexpression of Raf1 dominant-negative mutants inhibited IL-17F-induced GMCSF expression. The combination of PD98059 and Raf1 kinase inhibitor I completely blocked GM-CSF production, whereas 2 protein kinase C inhibitors, Ro-31-7549 and GF109203X, and a phosphatidylinositol 3-kinase inhibitor, LY294002, showed no inhibitory effect. Conclusion These findings suggest that ML-1 induces GM-CSF expression through the activation of the Raf1-MEK-extracellular signal-regulated kinase 1/2 pathway.
AB - Background ML-1 (IL-17F) is a recently discovered cytokine, and its function remains elusive. GM-CSF is a crucial cytokine for the maturation of various cell types and regulates allergic airway inflammation. Objective The functional effect of ML-1 in the expression of GM-CSF was investigated. Methods The levels of gene and protein expression in normal human bronchial epithelial cells (NHBEs) in the presence or absence of various kinase inhibitors or, in some cases, of a Raf1 dominant-negative mutant were determined by RT-PCR and ELISA, respectively. Western blotting was performed to investigate kinase activation. Results The results showed first that ML-1 induces, in a time-dependent and dose-dependent manner, the gene and protein expression for GM-CSF NHBEs, which are associated with activation of Raf1 and MAP kinase kinase (MEK) kinases. Selective MEK inhibitors, PD98059 and U0126, and Raf1 kinase inhibitor I significantly inhibited ML-1-induced GM-CSF production. Furthermore, overexpression of Raf1 dominant-negative mutants inhibited IL-17F-induced GMCSF expression. The combination of PD98059 and Raf1 kinase inhibitor I completely blocked GM-CSF production, whereas 2 protein kinase C inhibitors, Ro-31-7549 and GF109203X, and a phosphatidylinositol 3-kinase inhibitor, LY294002, showed no inhibitory effect. Conclusion These findings suggest that ML-1 induces GM-CSF expression through the activation of the Raf1-MEK-extracellular signal-regulated kinase 1/2 pathway.
KW - Bronchial epithelial cells
KW - Dimethyl sulfoxide
KW - DMSO
KW - ERK
KW - ERK1/2
KW - Extracellular signal-regulated kinase
KW - GM-CSF
KW - MAP kinase
KW - MAP kinase kinase
KW - MEK
KW - NF-κB
KW - NHBE
KW - Normal human bronchial epithelial cell
KW - Nuclear factor κB
KW - PI3K
KW - Raf1
UR - http://www.scopus.com/inward/record.url?scp=4344609710&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=4344609710&partnerID=8YFLogxK
U2 - 10.1016/j.jaci.2004.03.047
DO - 10.1016/j.jaci.2004.03.047
M3 - Article
C2 - 15316530
AN - SCOPUS:4344609710
VL - 114
SP - 444
EP - 450
JO - Journal of Allergy and Clinical Immunology
JF - Journal of Allergy and Clinical Immunology
SN - 0091-6749
IS - 2
ER -