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The p38 mitogen-activated kinase pathway regulates the human interleukin-10 promoter via the activation of Sp1 transcription factor in lipopolysaccharide-stimulated human macrophages

J Biol Chem. 2001 Apr 27;276(17):13664-74. doi: 10.1074/jbc.M011157200. Epub 2001 Jan 26.

Abstract

Interleukin-10 (IL-10), a pleiotropic cytokine that inhibits inflammatory and cell-mediated immune responses, is produced by a wide variety of cell types including T and B cells and monocytes/macrophages. Regulation of pro- and anti-inflammatory cytokines has been suggested to involve distinct signaling pathways. In this study, we investigated the regulation of the human IL-10 (hIL-10) promoter in the human monocytic cell line THP-1 following activation with lipopolysaccharide (LPS). Analysis of hIL-10 promoter sequences revealed that DNA sequences located between base pairs -652 and -571 are necessary for IL-10 transcription. A computer analysis of the promoter sequence between base pairs -652 and -571 revealed the existence of consensus sequences for Sp1, PEA1, YY1, and Epstein-Barr virus-specific nuclear antigen-2 (EBNA-2)-like transcription factors. THP-1 cells transfected with a plasmid containing mutant Sp1 abrogated the promoter activity, whereas plasmids containing the sequences for PEA1, YY1, and EBNA-2-like transcription factors did not influence hIL-10 promoter activity. To understand the events upstream of Sp1 activation, we investigated the role of p38 and extracellular signal-regulated kinase mitogen-activated protein kinases by using their specific inhibitors. SB202190 and SB203580, the p38-specific inhibitors, inhibited LPS-induced IL-10 production. In contrast, PD98059, a specific inhibitor of extracellular signal-regulated kinase kinases, failed to modulate IL-10 production. Furthermore, SB203580 inhibited LPS-induced activation of Sp1, as well as the promoter activity in cells transfected with a plasmid containing the Sp1 consensus sequence. These results suggest that p38 mitogen-activated protein kinase regulates LPS-induced activation of Sp1, which in turn regulates transcription of the hIL-10 gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins*
  • Base Sequence
  • Binding Sites
  • Cell Line
  • DNA-Binding Proteins / genetics
  • Enzyme Inhibitors / pharmacology
  • Enzyme-Linked Immunosorbent Assay
  • Epstein-Barr Virus Nuclear Antigens / genetics
  • Erythroid-Specific DNA-Binding Factors
  • Flavonoids / pharmacology
  • Flow Cytometry
  • Gene Expression Regulation, Enzymologic*
  • Genes, Reporter
  • Humans
  • Imidazoles / pharmacology
  • Interleukin-10 / biosynthesis
  • Interleukin-10 / genetics*
  • Interleukin-10 / metabolism*
  • Lipopolysaccharide Receptors / metabolism
  • Lipopolysaccharides / metabolism*
  • Luciferases / metabolism
  • Macrophages / metabolism*
  • Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • Mitogen-Activated Protein Kinases / metabolism*
  • Molecular Sequence Data
  • Plant Proteins / genetics
  • Plasmids / metabolism
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic*
  • Pyridines / pharmacology
  • Signal Transduction
  • Sp1 Transcription Factor / metabolism*
  • Time Factors
  • Transcription Factors / genetics
  • Transcription, Genetic
  • Transfection
  • Viral Proteins
  • YY1 Transcription Factor
  • p38 Mitogen-Activated Protein Kinases

Substances

  • Arabidopsis Proteins
  • DNA-Binding Proteins
  • EBNA-2 protein, Human herpesvirus 4
  • Enzyme Inhibitors
  • Epstein-Barr Virus Nuclear Antigens
  • Erythroid-Specific DNA-Binding Factors
  • Flavonoids
  • Imidazoles
  • Lipopolysaccharide Receptors
  • Lipopolysaccharides
  • PEA1 protein, Arabidopsis
  • Plant Proteins
  • Pyridines
  • Sp1 Transcription Factor
  • Transcription Factors
  • Viral Proteins
  • YY1 Transcription Factor
  • YY1 protein, human
  • Interleukin-10
  • Luciferases
  • Mitogen-Activated Protein Kinases
  • p38 Mitogen-Activated Protein Kinases
  • SB 203580
  • 4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one