Abstract | BACKGROUND: OBJECTIVE: We first confirmed that dsRNA up-regulated RANTES mRNA and protein synthesis in the airway epithelial cells. We next focused our studies on the transcriptional regulation of RANTES. METHODS: Airway epithelial cell line BEAS-2B and normal human bronchial epithelial cells were used in vitro study. Levels of RANTES mRNA and protein expression were determined with RT-PCR and ELISA. Mechanisms of transcriptional regulation were assessed by electrophoretic mobility shift assay and dual luciferase assay using RANTES promoter- luciferase reporter plasmids. RESULTS: Activation of nuclear factor-kappaB ( NF-kappaB) was confirmed by nuclear protein binding to a DNA probe derived from the RANTES promoter. Activity of the RANTES promoter was increased by dsRNA. The stimulation with dsRNA was partially inhibited in plasmids mutated at either of the binding sites for NF-kappaB or IFN regulatory factors (IRFs). When both sites were mutated, the activation was totally abrogated. CONCLUSION:
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Authors | K Ieki, S Matsukura, F Kokubu, T Kimura, H Kuga, M Kawaguchi, M Odaka, S Suzuki, S Watanabe, H Takeuchi, R P Schleimer, M Adachi |
Journal | Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology
(Clin Exp Allergy)
Vol. 34
Issue 5
Pg. 745-52
(May 2004)
ISSN: 0954-7894 [Print] England |
PMID | 15144466
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Chemokine CCL5
- Membrane Glycoproteins
- NF-kappa B
- RNA, Double-Stranded
- RNA, Messenger
- Receptors, Cell Surface
- TLR3 protein, human
- Toll-Like Receptor 3
- Toll-Like Receptors
- Interferons
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Topics |
- Bronchi
(metabolism)
- Cell Line
- Chemokine CCL5
(analysis, genetics)
- Electrophoretic Mobility Shift Assay
- Enzyme-Linked Immunosorbent Assay
(methods)
- Epithelial Cells
(metabolism)
- Gene Expression Regulation
- Humans
- Interferons
(metabolism)
- Membrane Glycoproteins
(genetics, metabolism)
- NF-kappa B
(metabolism)
- RNA, Double-Stranded
(physiology)
- RNA, Messenger
(analysis)
- Receptors, Cell Surface
(genetics, metabolism)
- Reverse Transcriptase Polymerase Chain Reaction
- Toll-Like Receptor 3
- Toll-Like Receptors
- Transcription, Genetic
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