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The Journal of Immunology, 2003, 170: 3099-3108.
Copyright © 2003 by The American Association of Immunologists

CD40-Mediated Transcriptional Regulation of the IL-6 Gene in B Lymphocytes: Involvement of NF-{kappa}B, AP-1, and C/EBP1

Mekhine Baccam*, So-Youn Woo*, Charles Vinson§ and Gail A. Bishop2,*,{dagger},{ddagger}

Departments of * Microbiology and {dagger} Internal Medicine, University of Iowa, and {ddagger} Veterans Affairs Medical Center, Iowa City, IA 52242; and § Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892

Engagement of CD40 by its ligand CD154 induces IL-6 production by B lymphocytes. We previously reported that this IL-6 production is dependent upon binding of the adapter protein TNF receptor-associated factor 6 to the cytoplasmic domain of CD40, while binding of TNF receptor-associated factors 2 and 3 is dispensable, as is the activation-induced nuclear translocation of NF-{kappa}B. The present study was designed to characterize CD40-mediated transcriptional control of the IL-6 gene in B cells. CD40 engagement on B lymphocytes activated the IL-6 promoter, and mutations in the putative binding sites for AP-1 and C/EBP transcription factors reduced this activation. Interestingly, a mutation in the putative NF-{kappa}B binding site completely abrogated the basal promoter activity, thus also rendering the promoter unresponsive to CD40 stimulation, suggesting that this site is required for binding of NF-{kappa}B constitutively present in the nucleus of mature B cells. The expression of dominant negative Fos or C/EBP{alpha} proteins, which prevent binding of AP-1 or C/EBP complexes to DNA, also reduced CD40-mediated IL-6 gene expression. Furthermore, CD40 stimulation led to phosphorylation of c-Jun on its activation domain, implicating CD40-mediated Jun kinase activation in the transcriptional regulation of IL-6 production.




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