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Molecular & Cellular Proteomics 5:1484-1496, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Comparative Immunoproteomics of Identification and Characterization of Virulence Factors from Helicobacter pylori Related to Gastric Cancer ^{* ,S}

Yu-Fen Lin, Ming-Shiang Wu^,¶, Chia-Che Chang^||, Sheng-Wei Lin, Jaw-Town Lin^,¶, Yuh-Ju Sun^**, Ding-Shinn Chen and Lu-Ping Chow^,,

From the Graduate Institute of Biochemistry and Molecular Biology and ^¶ Department of Primary Care Medicine, College of Medicine, National Taiwan University and Departments of Internal Medicine and Medical Genetics, National Taiwan University Hospital, Taipei 100, Taiwan, ^|| Institute of Biomedical Sciences, National Chung Hsing University, Taichung 402, Taiwan, and ^** Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 300, Taiwan

Helicobacter pylori is an important risk factor of gastric cancer (GC). Although many H. pylori virulence factors have been reported, the pathogenic mechanism by which H. pylori infection causes GC remains unclear. The aims of this study were to identify GC-related antigens from H. pylori and characterize their roles in the development of GC. As GC and duodenal ulcer (DU) are considered clinically divergent, we compared two-dimensional immunoblots of an acid-glycine extract of H. pylori probed with serum samples from 15 patients with GC and 15 with DU to find GC-related antigens, which were subsequently identified by mass spectrometry. Many protein spots were recognized by more than one serum, and 24 of these were better recognized by GC sera. The proteins showing higher frequency of recognition in GC group are threonine synthase, rod shape-determining protein, S-adenosylmethionine synthetase, peptide chain release factor 1, DNA-directed RNA polymerase subunit, co-chaperonin GroES (monomeric and dimeric forms), response regulator OmpR, and membrane fusion protein. Of these proteins, GroES was identified as a dominant GC-related antigen with a much higher seropositivity of GC samples (64.2%, n = 95) compared with 30.9% for gastritis (n = 94) and 35.5% for DU (n = 124). GroES seropositivity was more commonly associated with antral GC than with non-antral GC (odds ratio = 2.7; 95% confidence interval, 1.1–6.7). In peripheral blood mononuclear cells, GroES stimulated production of interleukin (IL)-8, IL-6, granulocyte macrophage colony-stimulating factor, IL-1ß, tumor necrosis factor-, cyclooxygenase-2, and prostaglandin E₂. Moreover when incubated with gastric epithelial cells, GroES induced expression of IL-8, cell proliferation, and up-regulation of c-jun, c-fos, and cyclin D1 but caused down-regulation of p27^Kip1. We conclude that GroES of H. pylori is a novel GC-associated virulence factor and may contribute to gastric carcinogenesis via induction of inflammation and promotion of cell proliferation.

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