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Divisions of Molecular Virology and Oncology,1 Control and Prevention of Infectious Diseases, Graduate School of Medicine,2 Divisions of Child Health and Welfare,3 Endocrinology and Metabolism, Faculty of Medicine,4 Department of Endoscopy, University Hospital, University of the Ryukyus, Nishihara, Okinawa 903-0215, Japan,7 Department of Pathology, Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8523, Japan,5 Division of Cytokine Signaling, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8523, Japan6
Received 30 May 2007/ Returned for modification 31 July 2007/ Accepted 16 August 2007
CCL20 attracts immature dendritic cells and memory T cells and plays a role on mucosal surfaces in inflammation. However, whether Helicobacter pylori infection induces CCL20 in human gastric epithelial cells remains to be determined. The aim of this study was to analyze the molecular mechanism of H. pylori-induced CCL20 expression. Expression of CCL20 mRNA was assessed by reverse transcription-PCR. Five normal and five H. pylori-infected gastric tissue samples were stained immunohistochemically for CCL20. A luciferase assay was used to monitor activation of the CCL20 gene promoter, and an electrophoretic mobility shift assay was used to explore the binding of transcription factors to this promoter. The CCL20 expression in epithelial cells of H. pylori-positive tissues was higher than that in H. pylori-negative tissues. H. pylori induced CCL20 expression in gastric epithelial cell lines, and the induction was dependent on an intact cag pathogenicity island. Activation of the CCL20 promoter by H. pylori occurred through the action of NF-
B. Transfection of I
B kinase and NF-
B-inducing kinase dominant negative mutants inhibited H. pylori-mediated activation of CCL20. Treatment with an inhibitor of Hsp90 suppressed H. pylori-induced CCL20 mRNA due to deactivation of NF-
B. Collectively, these results suggest that H. pylori activates NF-
B through an intracellular signaling pathway that involves I
B kinase and NF-
B-inducing kinase, leading to CCL20 gene transcription, and that Hsp90 is a crucial regulator of H. pylori-induced CCL20 expression, presumably contributing to the immune response in H. pylori.
Published ahead of print on 27 August 2007.
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