Abstract
The molecular mechanisms that regulate expression of the immunosuppressive cytokine IL-10 remain poorly understood. In this study, by measuring sensitivity to DNase I digestion, we show that production of IL-10 by primary mouse bone marrow-derived macrophages stimulated through pattern recognition receptors was associated with chromatin remodeling of the IL-10 locus. We also demonstrate that the IL-10 locus is remodeled in primary Th2 cells and IL-10-producing regulatory T cells that have been differentiated in vitro. Strikingly, a novel DNase I-hypersensitive site (HSS-4.5) was identified in stimulated macrophages, but not in T cells. We show that hyperacetylated histones were recruited to this site in stimulated macrophages. Furthermore, HSS-4.5 is highly conserved and contains a putative NF-kappaB binding site. In support of a function for this site, NF-kappaB p65/RelA was recruited to HSS-4.5 in vivo and its activation was required for optimal IL-10 gene expression in LPS-stimulated macrophages.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Base Sequence
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Bone Marrow Cells / enzymology
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Bone Marrow Cells / immunology
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Bone Marrow Cells / metabolism
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Cell Differentiation / genetics
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Cell Differentiation / immunology
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Chromatin / genetics
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Chromatin / immunology*
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Chromatin / metabolism*
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Dendritic Cells / enzymology
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Dendritic Cells / immunology
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Dendritic Cells / metabolism
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Deoxyribonuclease I / genetics
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Deoxyribonuclease I / metabolism
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Genetic Markers / immunology
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Humans
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Interleukin-10 / biosynthesis*
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Interleukin-10 / genetics*
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Interleukin-10 / metabolism
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Lipopolysaccharides / pharmacology
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Macrophage Activation / genetics
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Macrophage Activation / immunology
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Macrophages / enzymology
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Macrophages / immunology*
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Macrophages / metabolism*
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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NF-kappa B / biosynthesis
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NF-kappa B / metabolism
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Protein Transport / genetics
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Protein Transport / immunology
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Regulatory Sequences, Nucleic Acid / immunology
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Resting Phase, Cell Cycle / genetics
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Resting Phase, Cell Cycle / immunology
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T-Lymphocytes, Regulatory / immunology
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T-Lymphocytes, Regulatory / metabolism
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Th2 Cells / cytology
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Th2 Cells / immunology
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Th2 Cells / metabolism
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Transcription Factor RelA
Substances
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Chromatin
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Genetic Markers
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Lipopolysaccharides
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NF-kappa B
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Transcription Factor RelA
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Interleukin-10
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Deoxyribonuclease I