Most virus-infected cells release interferon-β (IFN-β) as a powerful inducer of antiviral defense. Endothelial cells tightly regulate local immune cell recruitment by expression of adhesion molecules and chemokines. Here, we studied the transcriptional regulation of IFN-β-induced chemokine expression in primary human endothelial cells. IFN-β moderately increased monocyte chemoattractant protein-1/CCL2 and potently raised IFN-γ-inducible protein-10/CXCL10 mRNA steady-state levels and protein release, while no effect was detected on various other chemokines. As shown by transient transfections, induction of CXCL10 expression depends on an IFN-stimulated response element (ISRE) within the CXCL10 promoter. A double point mutation of the putative IFN regulatory factor (IRF)-1/2 binding site within this ISRE motif abolished IFN-β-induced promoter activity. In electrophoretic mobility shift assays, this ISRE motif showed a basal IRF-2 and an IFN-β-inducible IRF-1 and augmented IRF-2 binding. Furthermore, stimulation with IFN-β induced a rapid nuclear translocation of signal transducer and activator of transcription 1 (STAT1) and STAT2 and their transient binding to a γ-activated site within the CCL2 promoter. The kinetics of transient STAT1 binding to this γ-activated site element correlated with the amount of Y701-phosphorylated nuclear STAT1, while S727-phosphorylated nuclear STAT1 remained stable over 24 h after stimulation. Therefore, IFN-β potently induces endothelial chemokine expression at the transcriptional level.

Keywords:
Endothelial cells, Interferon-β, CXC chemokine IP-10, Monocyte chemoattractant protein-1, Interferon regulatory factor-1, Interferon regulatory factor-2, Interferon-stimulated gene factor 3 complex, γ-Activated factor, Signal transducer and activator of transcription 2 protein, Multiple sclerosis
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