Identification of a macrophage-specific chromatin signature in the IL-10 locus.
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.
Divisions of Immunoregulation, Immune Cell Biology and Molecular Immunology, National Institute for Medical Research, The Ridgeway, London NW7 1AA, United Kingdom. msaraiv@nimr.mrc.ac.uk
Margarida
M
Saraiva
Jillian R
JR
Christensen
Alla V
AV
Tsytsykova
Anne E
AE
Goldfeld
Steven C
SC
Ley
Dimitris
D
Kioussis
Anne
A
O'Garra
D000818:Animals; D001483:Base Sequence; D001854:Bone Marrow Cells / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology; D002843:Chromatin / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D003713:Dendritic Cells / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000235:genetics / Q000378:metabolism; D005819:Genetic Markers / Q000276:immunology; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008262:Macrophage Activation / Q000235:genetics / Q000276:immunology; D008264:Macrophages / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000096:biosynthesis / Q000378:metabolism; D021381:Protein Transport / Q000235:genetics / Q000276:immunology; D012045:Regulatory Sequences, Nucleic Acid / Q000276:immunology; D016192:Resting Phase, Cell Cycle / Q000235:genetics / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology / Q000378:metabolism; D018418:Th2 Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D051996:Transcription Factor RelA
D002843:Chromatin; D005819:Genetic Markers; D008070:Lipopolysaccharides; D016328:NF-kappa B; D051996:Transcription Factor RelA; D016753:Interleukin-10; D003850:Deoxyribonuclease I