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CD40 induces interleukin-6 gene transcription in dendritic cells: regulation by TRAF2, AP-1, NF-?B, AND CBF1

CD40 induces interleukin-6 gene transcription in dendritic cells: regulation by TRAF2, AP-1, NF-?B, AND CBF1
CD40 induces interleukin-6 gene transcription in dendritic cells: regulation by TRAF2, AP-1, NF-?B, AND CBF1
CD40-induced activation of cytokine gene expression in dendritic cells (DC) is an important process in the initiation of primary immune responses. We have determined the intracellular signaling events that lead to CD40 ligation-induced activation of interleukin-6 (IL-6) gene transcription in a murine DC line, FSDC, that is phenotypically representative of bone marrow-derived DC. IL-6 reverse transcriptase-PCR and promoter assays established the responsiveness of FSDC to anti-CD40 ligation. Further promoter assays showed that the transcription factors NF-?B and AP-1 are downstream transcriptional mediators of CD40-induced IL-6 gene expression. Anti-CD40 treatment of FSDC stimulated increased expression of specific NF-?B (p50:p65) and AP-1 (c-Jun, JunB, JunD, and c-Fos) DNA-protein complexes. Overexpression of an I?B-? super-repressor or a dominant negative JunD resulted in a strong inhibition of CD40-inducible IL-6 promoter activity supporting a role for both transcription factors. Upstream signal transduction events were studied by transfection of wild type and mutant human CD40 expression constructs into FSDC followed by stimulation with an anti-human CD40 antibody. These experiments revealed that anti-CD40 stimulation of NF-?B and IL-6 gene transcription requires specific amino acid residues in the cytoplasmic region of CD40 involved in the recruitment of TRAF2. Induction of IL-6 mRNA by anti-CD40 treatment was found to be a transient event (24 h) and was followed by a diminution of IL-6 transcript to levels below those found in unstimulated cells. This loss of IL-6 expression was associated with reduced p50:p65 NF-?B DNA binding and elevated binding of CBF1 to a site overlapping the NF-?B site. Overexpression of CBF1 resulted in a profound inhibition of basal and anti-CD40-induced IL-6 promoter activities indicating that prolonged induction of CBF1 may contribute to the transient nature of the IL-6 response. The physiological relevance of these molecular events to DC function is discussed.
0021-9258
17125-17138
Mann, Jelena
23defaa6-4d36-4d53-909c-f1375ee73df8
Oakley, Fiona
f5f32319-966d-46b1-b774-bd7548022ff3
Johnson, Peter W.M.
3f6068ce-171e-4c2c-aca9-dc9b6a37413f
Mann, Derek A.
7a0eb3d7-c2ca-432f-82a0-f4b0df08755d
Mann, Jelena
23defaa6-4d36-4d53-909c-f1375ee73df8
Oakley, Fiona
f5f32319-966d-46b1-b774-bd7548022ff3
Johnson, Peter W.M.
3f6068ce-171e-4c2c-aca9-dc9b6a37413f
Mann, Derek A.
7a0eb3d7-c2ca-432f-82a0-f4b0df08755d

Mann, Jelena, Oakley, Fiona, Johnson, Peter W.M. and Mann, Derek A. (2002) CD40 induces interleukin-6 gene transcription in dendritic cells: regulation by TRAF2, AP-1, NF-?B, AND CBF1. The Journal of Biological Chemistry, 277 (19), 17125-17138. (doi:10.1074/jbc.M109250200).

Record type: Article

Abstract

CD40-induced activation of cytokine gene expression in dendritic cells (DC) is an important process in the initiation of primary immune responses. We have determined the intracellular signaling events that lead to CD40 ligation-induced activation of interleukin-6 (IL-6) gene transcription in a murine DC line, FSDC, that is phenotypically representative of bone marrow-derived DC. IL-6 reverse transcriptase-PCR and promoter assays established the responsiveness of FSDC to anti-CD40 ligation. Further promoter assays showed that the transcription factors NF-?B and AP-1 are downstream transcriptional mediators of CD40-induced IL-6 gene expression. Anti-CD40 treatment of FSDC stimulated increased expression of specific NF-?B (p50:p65) and AP-1 (c-Jun, JunB, JunD, and c-Fos) DNA-protein complexes. Overexpression of an I?B-? super-repressor or a dominant negative JunD resulted in a strong inhibition of CD40-inducible IL-6 promoter activity supporting a role for both transcription factors. Upstream signal transduction events were studied by transfection of wild type and mutant human CD40 expression constructs into FSDC followed by stimulation with an anti-human CD40 antibody. These experiments revealed that anti-CD40 stimulation of NF-?B and IL-6 gene transcription requires specific amino acid residues in the cytoplasmic region of CD40 involved in the recruitment of TRAF2. Induction of IL-6 mRNA by anti-CD40 treatment was found to be a transient event (24 h) and was followed by a diminution of IL-6 transcript to levels below those found in unstimulated cells. This loss of IL-6 expression was associated with reduced p50:p65 NF-?B DNA binding and elevated binding of CBF1 to a site overlapping the NF-?B site. Overexpression of CBF1 resulted in a profound inhibition of basal and anti-CD40-induced IL-6 promoter activities indicating that prolonged induction of CBF1 may contribute to the transient nature of the IL-6 response. The physiological relevance of these molecular events to DC function is discussed.

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Published date: 2002

Identifiers

Local EPrints ID: 26460
URI: http://eprints.soton.ac.uk/id/eprint/26460
ISSN: 0021-9258
PURE UUID: 52c9d1f9-df18-4a2a-9656-cb699d14888d
ORCID for Peter W.M. Johnson: ORCID iD orcid.org/0000-0003-2306-4974

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Date deposited: 21 Apr 2006
Last modified: 20 Jul 2019 01:12

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