Transcriptional Regulation of Deoxynivalenol-Induced IL-8 Expression in Human Monocytes

• Published in: Toxicological sciences 2007-10, Vol.99 (2), p.502-511
• Main Authors: Gray, Jennifer S., Pestka, James J.
• Format: Article
• Language: English
• Subjects: CCAAT-Enhancer-Binding Protein-beta - physiology; Cells, Cultured; chemokine; Gene Expression Regulation - drug effects; Humans; immunotoxicity; Interleukin-8 - genetics; Monocytes - metabolism; mycotoxin; NF-kappa B - physiology; p38 Mitogen-Activated Protein Kinases - physiology; RNA Stability; transcription; Transcription Factor RelA - metabolism; Transcription, Genetic; trichothecene; Trichothecenes - pharmacology
• ISSN: 1096-6080; 1096-0929
• DOI: 10.1093/toxsci/kfm182

The trichothecene mycotoxin deoxynivalenol (DON), commonly present in contaminated grains worldwide, induces expression of the chemokine interleukin (IL)-8 in human monocytes. The purpose of this study was to test the hypothesis that DON modulates transcriptional and posttranscriptional regulation of IL-8 expression in the U937 human monocyte model. When U937 cells were transfected with a wild-type IL-8 promoter luciferase construct (−162/+44 IL-8 LUC) and incubated with DON (1 μg/ml) or the positive control, lipopolysaccharide (LPS) (1 μg/ml), there was a significant increase in luciferase expression. Mutation of the nuclear factor-κB (NF-κB) binding site significantly impaired both DON- and LPS-induced luciferase expression. In contrast, mutating the activator protein-1 binding site resulted in significantly increased DON- and LPS-induced luciferase expression. CCAAT/enhancer-binding protein β, octamer-1, or NF-κB repressing factor binding site mutations did not affect DON-induced luciferase activity. Consistent with reporter studies, the NF-κB inhibitor caffeic acid phenethyl ester completely ablated both DON-induced IL-8 mRNA and protein expression. When NF-κB subunit binding to a specific IL-8 promoter probe was evaluated by enzyme-linked immunosorbent assay (ELISA), DON was observed to increase p65 binding by 21-fold, have no effect on p50 binding and decrease p52 binding. DON was not found to stabilize IL-8 mRNA in U937 cells. Taken together, these data suggest that DON-induced IL-8 expression is likely to be mediated at the transcriptional level by NF-κB, specifically p65, but does not appear to involve mRNA stabilization.