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Mechanism of interferon-gamma mediated down-regulation of interleukin-10 gene expression

Mechanism of interferon-gamma mediated down-regulation of interleukin-10 gene expression
Mechanism of interferon-gamma mediated down-regulation of interleukin-10 gene expression
Expression of the anti-inflammatory cytokine IL-10 is suppressed by the pro-inflammatory interferon gamma but the mechanism of this action is unknown. We analysed activity of IL-10 promoter luciferase reporter constructs in transfected RPMI 8226.1 B cells that were treated at -2 h with IFNgamma (1000 U/ml) followed by stimulation with LPS (100 ng/ml) at 0 h. IFNgamma treatment suppressed LPS-induced IL-10 promoter activity in a construct carrying the -1044 promoter and also one containing the -195 promoter. The suppression was independent of the IRF-motif at -182 but involved the Stat-motif at -120. In gelshift analysis this Stat motif did bind LPS-induced Stat3 and with IFNgamma treatment it did, in addition, bind Stat1. ChIP analysis for detection of transcription factor binding to chromatin in intact cells demonstrated Stat3 binding to the proximal IL-10 promoter when cells are stimulated with LPS only. Treatment with IFNgamma only led to Stat1 binding in ChIP analysis and treatment with IFNgamma plus LPS led to reduced Stat3 binding while Stat1 binding remained high. Finally, LPS-induced activity of the trimeric Stat-motif in front of the luciferase reporter was suppressed by IFNgamma. These data demonstrate that IFNgamma down-regulates expression of the IL-10 gene by a novel mechanism that involves displacement of transactiving Stat3 by IFNgamma-induced Stat1.
0161-5890
1351-1359
Schaefer, A.
5f9e7a2a-328f-4031-acf5-be6023aea20d
Unterberger, C.
ce704436-9e9b-48b4-9a52-1a302ed98172
Frankenberger, M.
b6dbd3a3-5dd4-4008-bb5b-e834476f85fa
Lohrum, Marion
a358dc72-0837-4d18-b431-dbf9a5f9a439
Staples, Karl J.
e0e9d80f-0aed-435f-bd75-0c8818491fee
Werner, Thomas
b7ef6029-1dcf-4080-b952-a9fca65c1873
Stunnenberg, Henk
a7b7ea55-cac3-4761-9dba-18ce0e834f09
Ziegler-Heitbrock, Loems
71bb9dd4-262c-46ba-9c10-1be5941ffca4
Schaefer, A.
5f9e7a2a-328f-4031-acf5-be6023aea20d
Unterberger, C.
ce704436-9e9b-48b4-9a52-1a302ed98172
Frankenberger, M.
b6dbd3a3-5dd4-4008-bb5b-e834476f85fa
Lohrum, Marion
a358dc72-0837-4d18-b431-dbf9a5f9a439
Staples, Karl J.
e0e9d80f-0aed-435f-bd75-0c8818491fee
Werner, Thomas
b7ef6029-1dcf-4080-b952-a9fca65c1873
Stunnenberg, Henk
a7b7ea55-cac3-4761-9dba-18ce0e834f09
Ziegler-Heitbrock, Loems
71bb9dd4-262c-46ba-9c10-1be5941ffca4

Schaefer, A., Unterberger, C., Frankenberger, M., Lohrum, Marion, Staples, Karl J., Werner, Thomas, Stunnenberg, Henk and Ziegler-Heitbrock, Loems (2009) Mechanism of interferon-gamma mediated down-regulation of interleukin-10 gene expression. Molecular Immunology, 46 (7), 1351-1359. (doi:10.1016/j.molimm.2008.11.015). (PMID:19124156)

Record type: Article

Abstract

Expression of the anti-inflammatory cytokine IL-10 is suppressed by the pro-inflammatory interferon gamma but the mechanism of this action is unknown. We analysed activity of IL-10 promoter luciferase reporter constructs in transfected RPMI 8226.1 B cells that were treated at -2 h with IFNgamma (1000 U/ml) followed by stimulation with LPS (100 ng/ml) at 0 h. IFNgamma treatment suppressed LPS-induced IL-10 promoter activity in a construct carrying the -1044 promoter and also one containing the -195 promoter. The suppression was independent of the IRF-motif at -182 but involved the Stat-motif at -120. In gelshift analysis this Stat motif did bind LPS-induced Stat3 and with IFNgamma treatment it did, in addition, bind Stat1. ChIP analysis for detection of transcription factor binding to chromatin in intact cells demonstrated Stat3 binding to the proximal IL-10 promoter when cells are stimulated with LPS only. Treatment with IFNgamma only led to Stat1 binding in ChIP analysis and treatment with IFNgamma plus LPS led to reduced Stat3 binding while Stat1 binding remained high. Finally, LPS-induced activity of the trimeric Stat-motif in front of the luciferase reporter was suppressed by IFNgamma. These data demonstrate that IFNgamma down-regulates expression of the IL-10 gene by a novel mechanism that involves displacement of transactiving Stat3 by IFNgamma-induced Stat1.

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Published date: April 2009
Organisations: Clinical & Experimental Sciences

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Local EPrints ID: 360138
URI: http://eprints.soton.ac.uk/id/eprint/360138
ISSN: 0161-5890
PURE UUID: e35b2fc9-abf7-4d40-8167-4a65ec5fa366
ORCID for Karl J. Staples: ORCID iD orcid.org/0000-0003-3844-6457

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Date deposited: 27 Nov 2013 13:33
Last modified: 15 Mar 2024 03:27

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Contributors

Author: A. Schaefer
Author: C. Unterberger
Author: M. Frankenberger
Author: Marion Lohrum
Author: Karl J. Staples ORCID iD
Author: Thomas Werner
Author: Henk Stunnenberg
Author: Loems Ziegler-Heitbrock

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