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Nuclear factor-κB does not mediate the inhibitory effects of dexamethasone on granulocyte–macrophage colony-stimulating factor expression

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Summary

Human granulocyte–macrophage colony-stimulating factor (GM-CSF) reporter constructs containing up to 3ยท3 kb of upstream promoter sequence were transiently transfected into both Jurkat and HUT78 human T-cell lines. In Jurkat cells, stimulation with phorbol 12-myristate 13-acetate (PMA) plus phytohemaglutinin (PHA) produced robust increases in reporter activity, whereas HUT78 cells showed low levels of reporter induction attributable to constitutive nuclear factor (NF)-κB activity. Following mutation of either the proximal NF-κB site (−85/−76) or the activator protein1 (AP-1) motif within the conserved lymphokine element 0 (CLE0) site (−54/−31), reporter activity was markedly reduced in both cell lines. Despite this dependence on NF-κB and CLE0/AP-1, GM-CSF reporter activity was unaffected by dexamethasone in either cell line. Similarly, an NF-κB-dependent reporter was also not repressed by dexamethasone, yet GM-CSF release from HUT78 T cells was inhibited. These data therefore confirm a critical role for both NF-κB and CLE0 sites in GM-CSF promoter activation and indicate that NF-κB may not mediate glucocorticoid-dependent repression of GM-CSF in these cells.
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Document Type: Research Article

Affiliations: Department of Thoracic Medicine, National Heart & Lung Institute, Faculty of Medicine, Imperial College London, London, UK

Publication date: April 1, 2004

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