title: Interleukin-10 (IL-10) mediated suppression of IL-12 production in RAW 264.7 cells also involves c-rel transcription factor.
abstract: Interleukin-10 (IL-10) is known to inhibit IL-12 production in macrophages primarily at the transcriptional level with the involvement of p50 and p65 nuclear factor-kappaB (NF-kappaB). We demonstrate that the c-rel transcription factor also plays a major role in IL-10-mediated IL-12 suppression. Treatment of macrophages with recombinant IL-10 inhibited nuclear c-rel levels, whereas addition of neutralizing anti-IL-10 antibody up-regulated both nuclear c-rel levels and IL-12 production by macrophages. Decreased nuclear c-rel was associated with a reduction in phosphorylation of inhibitory kappa B alpha (IkappaBalpha) in the cytoplasm, indicating that IL-10 prevents degradation of IkappaBalpha and the subsequent translocation of c-rel into the nucleus. Treatment with leptomycin B, a known inhibitor of c-rel at a concentration of 10 nm, when used with anti-IL-10 antibody, resulted in reduced expression of IL-12. In a complementary experiment, in vitro transient expression of p65 NF-kappaB could not rescue the inhibitory effect of IL-10 on IL-12 production, suggesting that NF-kappaB alone was not sufficient to restore IL-12 production during IL-10 treatment. However, over-expression of c-rel resulted in IL-12 restoration upon stimulation with lipopolysaccharide plus interferon-gamma during IL-10 treatment. Our studies highlight the involvement of c-rel in IL-10-mediated IL-12 regulation.
authors.affiliation: Centre for DNA Fingerprinting and Diagnostics (CDFD), Nacharam, Hyderabad, India.
authors.forename: Sheikh Showkat
authors.initials: SS
authors.lastname: Rahim
authors.forename: Nooruddin
authors.initials: N
authors.lastname: Khan
authors.forename: Chandra Sekhar
authors.initials: CS
authors.lastname: Boddupalli
authors.forename: Seyed E
authors.initials: SE
authors.lastname: Hasnain
authors.forename: Sangita
authors.initials: S
authors.lastname: Mukhopadhyay
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism; D020672:I-kappa B Proteins / Q000378:metabolism; D016753:Interleukin-10 / Q000276:immunology / Q000494:pharmacology*; D018664:Interleukin-12 / Q000096:biosynthesis*; D008070:Lipopolysaccharides / Q000276:immunology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology*; D051379:Mice; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D020569:Proto-Oncogene Proteins c-rel / Q000037:antagonists & inhibitors / Q000502:physiology*; D011994:Recombinant Proteins / Q000494:pharmacology; D051996:Transcription Factor RelA; D015854:Up-Regulation / Q000276:immunology
chemical_list: D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D008070:Lipopolysaccharides; D016328:NF-kappa B; C000606205:Nfkbia protein, mouse; D020569:Proto-Oncogene Proteins c-rel; D011994:Recombinant Proteins; D051996:Transcription Factor RelA; D016753:Interleukin-10; D000072000:NF-KappaB Inhibitor alpha; D018664:Interleukin-12
references.citation: J Biol Chem. 1999 Nov 5;274(45):31868-74
references.pmid: 10542212
references.citation: Mol Cell Biol. 1999 Oct;19(10):7088-95
references.pmid: 10490645
references.citation: Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8427-32
references.pmid: 10890924
references.citation: Proc Natl Acad Sci U S A. 2000 Nov 7;97(23):12705-10
references.pmid: 11058167
references.citation: J Immunol. 2001 Feb 1;166(3):1823-31
references.pmid: 11160229
references.citation: J Exp Med. 2001 Oct 15;194(8):1021-32
references.pmid: 11602633
references.citation: J Immunol. 2002 Mar 15;168(6):2590-4
references.pmid: 11884420
references.citation: J Immunol. 2002 Mar 15;168(6):2914-21
references.pmid: 11884462
references.citation: J Biol Chem. 2003 Jan 31;278(5):2773-6
references.pmid: 12468543
references.citation: Nat Rev Immunol. 2003 Feb;3(2):133-46
references.pmid: 12563297
references.citation: Scand J Immunol. 2003 Jul;58(1):23-32
references.pmid: 12828555
references.citation: Clin Exp Immunol. 2004 Jan;135(1):64-73
references.pmid: 14678266
references.citation: FEBS Lett. 2004 Jun 4;567(2-3):275-80
references.pmid: 15178336
references.citation: Methods Enzymol. 1983;101:582-98
references.pmid: 6888276
references.citation: Proc Natl Acad Sci U S A. 1991 May 15;88(10):4143-7
references.pmid: 1674604
references.citation: J Exp Med. 1991 Nov 1;174(5):1209-20
references.pmid: 1940799
references.citation: J Exp Med. 1992 Nov 1;176(5):1387-98
references.pmid: 1357073
references.citation: J Immunol. 1995 May 15;154(10):5071-9
references.pmid: 7730613
references.citation: Mol Cell Biol. 1997 Aug;17(8):4572-88
references.pmid: 9234715
references.citation: Nature. 1997 Aug 7;388(6642):548-54
references.pmid: 9252186
references.citation: Semin Cancer Biol. 1997 Apr;8(2):63-73
references.pmid: 9299584
references.citation: Science. 1997 Oct 31;278(5339):860-6
references.pmid: 9346484
references.citation: J Immunol. 1998 Jun 15;160(12):5936-44
references.pmid: 9637507
references.citation: Mol Cell Biol. 1998 Sep;18(9):5445-56
references.pmid: 9710628
references.citation: J Immunol. 1999 Jul 15;163(2):875-83
references.pmid: 10395682
references.citation: J Immunol. 1999 Aug 15;163(4):1786-92
references.pmid: 10438910
references.citation: Proc Natl Acad Sci U S A. 2000 Feb 1;97(3):1014-9
references.pmid: 10655476