Abstract
CC chemokine receptor 1 (CCR1) has been implicated in inflammation. The present study examined the signaling mechanisms that mediate GM-CSF/IL-10-induced synergistic CCR1 protein expression in monocytic U937 cells. GM-CSF alone markedly increased both the mRNA and protein expression of CCR1. IL-10 augmented GM-CSF-induced CCR1 protein expression with no effect on mRNA expression. PD098059 and U0126 (two MEK inhibitors), and LY294002 (a PI3K inhibitor) inhibited GM-CSF/IL-10-induced CCR1 gene and protein expression. PD098059, U0126, and LY294002 also attenuated chemotaxis of GM-CSF/IL-10-primed U937 cells in response to MIP-1alpha. Immunoblotting studies show that GM-CSF alone induced ERK2 phosphorylation; whereas, IL-10 alone induced p70(S6k) phosphorylation in U937 cells. Neither cytokine when used alone induced PKB/Akt phosphorylation. Combined GM-CSF/IL-10 treatment of U937 cells induced phosphorylation of ERK2, p70(S6k), and PKB/Akt. PD098059 and U0126 completely abrogated ERK2 phosphorylation; whereas, LY294002 completely blocked PKB/Akt and p70(S6k) phosphorylation. Our findings indicate that IL-10 may potentiate GM-CSF-induced CCR1 protein expression in U937 cells via activation of PKB/Akt and p70(S6k).
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Chemokine CCL3
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Chemokine CCL4
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Chemotaxis, Leukocyte
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Drug Synergism
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Enzyme Inhibitors / 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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Interleukin-10 / pharmacology*
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Macrophage Inflammatory Proteins / pharmacology
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Mitogen-Activated Protein Kinases / metabolism
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Monocytes / drug effects
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Monocytes / immunology*
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Myeloid Cells / drug effects
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Myeloid Cells / immunology
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Phosphorylation
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Protein Serine-Threonine Kinases*
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Proto-Oncogene Proteins / metabolism
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Proto-Oncogene Proteins c-akt
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RNA, Messenger / biosynthesis
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Receptors, CCR1
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Receptors, Chemokine / biosynthesis*
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Receptors, Chemokine / genetics
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Ribosomal Protein S6 Kinases, 70-kDa / metabolism
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Signal Transduction / drug effects
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U937 Cells
Substances
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CCR1 protein, human
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Chemokine CCL3
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Chemokine CCL4
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Enzyme Inhibitors
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Macrophage Inflammatory Proteins
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Proto-Oncogene Proteins
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RNA, Messenger
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Receptors, CCR1
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Receptors, Chemokine
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Interleukin-10
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Granulocyte-Macrophage Colony-Stimulating Factor
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AKT1 protein, human
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Protein Serine-Threonine Kinases
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Proto-Oncogene Proteins c-akt
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Ribosomal Protein S6 Kinases, 70-kDa
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Mitogen-Activated Protein Kinases