Anti-Helicobacter pylori antibody succession during gastric carcinogenesis monitored with proteomics and antigen-chip analysis

• Project/Area Number: 24590919
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Yamaguchi University
• Principal Investigator: AKADA Junko 山口大学, 医学部, 特別医学研究員 (30346548)
• Co-Investigator(Kenkyū-buntansha): NISHIKAWA Jun 山口大学, 医学部附属病院, 准教授 (00379950) ; OKUDA Masumi 兵庫医科大学, 医学部, 教授 (40531091)
• Co-Investigator(Renkei-kenkyūsha): HIRAYAMA Toshiya 長崎大学, 熱帯医学研究所, 教授 (50050696) ; NAKAMURA Kazuyuki 山口大学, 名誉教授 (90107748) ; NAKAMURA Mikiko 山口大学, 大学研究推進機構, 学術研究員 (20457310)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: Helicobacter pylori / 血清 / 抗H. pylori抗体 / 抗原タンパク質 / CagA / 小児 / 血清抗体 / H. pylori / プロテオミクス / 抗原解析 / ヘリコバクター・ピロリ / 抗体

Outline of Final Research Achievements

This study focused on Japanese child serum antibodies against Helicobacter pylori, a chronically-infected gastric bacterium which causes gastric cancer in adults. Serum samples from 24 children, 22 H. pylori (Hp)-positive and 2 Hp-negative children, were used to catalogue antigenic proteins of a Japanese strain CPY2052 by two-dimensional electrophoresis followed by immunoblot and LC-MS/MS analysis. In total, 24 proteins were identified as candidate antigen proteins. Among these, the major virulence factor, cytotoxin-associated gene A (CagA) protein was the most reactive antigen recognized by all the Hp-positive sera even from children under the age of 3 years. The major antigenic part of CagA was identified in the middle region, and two peptides containing CagA epitopes were identified using a newly developed peptide/protein-combined array chip method. Each of the epitopes was found to contain amino acid residue(s) unique to East Asian CagA.

Research Products

• [Journal Article] Proteomic Characterization of Helicobacter pylori CagA Antigen Recognized by Child Serum Antibodies and Its Epitope Mapping by Peptide Array (2014)
  • Author(s): Junko Akada , Masumi Okuda, Narumi Hiramoto, Takao Kitagawa, Xiulian Zhang, Shuichi Kamei, Akane Ito, Mikiko Nakamura, Tomohisa Uchida, Tomoko Hiwatani, Yoshihiro Fukuda, Teruko Nakazawa, Yasuhiro Kuramitsu, Kazuyuki Nakamura
  • Journal Title: PLOS ONE Volume: 9 Issue: 8 Pages: e104611-e104611
  • DOI: 10.1371/journal.pone.0104611
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A new type of protein chip to detect hepatocellular carcinoma-related autoimmune antibodies in the sera of hepatitis C virus-positive patients. (2013)
  • Author(s): Akada J, Kamei S, Ito A, Ito M, Kitagawa T, Furumoto H, Kato Y, Tamesa M, Takashima M, Shirai M, Yamano H, Oka M, Kuramitsu Y, Nakamura K.
  • Journal Title: Proteome Sci. Volume: 11(1) Issue: 1 Pages: 33-33
  • DOI: 10.1186/1477-5956-11-33
  • Peer Reviewed
• [Presentation] Helicobacter pylori CagA as a dominant antigen recognized by Japanese child serum antibodies (2014)
  • Author(s): Masumi Okuda, Junko Akada, Kazuyuki Nakamura, Teruko Nakazawa, Yoshihiro Fukuda
  • Organizer: European Helicobacter Study Group International Workshop on Helicobacter & Microbiota in Inflammation & Cancer
  • Place of Presentation: Rome, Italy
  • Year and Date: 2014-09-10 – 2014-09-13
• [Presentation] ピロリ菌感染小児抗体による東アジアCagA特異的エピトープの同定 (2014)
  • Author(s): 赤田純子、奥田真珠美、福田能啓、中澤晶子、中村和行
  • Organizer: 日本ヘリコバクター学会
  • Place of Presentation: ステーションコンファレンス東京（東京都千代田区）
  • Year and Date: 2014-06-28 – 2014-06-29
• [Presentation] ピロリ菌感染小児血清中の抗ピロリ菌抗体群とCagA抗原ドメインの解析 (2013)
  • Author(s): 赤田純子、奥田真珠美、福田能啓、中澤晶子、中村和行
  • Organizer: 第１9回日本ヘリコバクター学会
  • Place of Presentation: 長崎(長崎大学医学部良順会館記念講堂）
  • Invited
• [Presentation] ピロリ菌感染小児血清中の抗ピロリ菌抗体群の解析 (2012)
  • Author(s): 赤田純子、奥田真珠美、内田智久、福田能啓、中澤晶子、中村和行
  • Organizer: 第１8回日本ヘリコバクター学会
  • Place of Presentation: 岡山（岡山コンベンションセンター）
• [Presentation] ピロリ菌感染小児血清中の抗ピロリ菌抗体群とCagA抗原ドメインの解析
  • Author(s): 赤田純子、奥田真珠美、福田能啓、中澤晶子、中村和行
  • Organizer: 日本ヘリコバクター学会
  • Place of Presentation: 長崎大学 良順会館 (長崎県）
  • Invited
• [Presentation] Immuno-proteomics of Helicobacter pylori antigens for IgG antibodies in H. pylori-positive Japanese child sera
  • Author(s): Junko Akada, Masumi Okuda, Yoshihiro Fukuda, Takao Kitagawa, Yasuhiro Kuramitsu, and Kazuyuki Nakamura
  • Organizer: HUPO 12th Annual World Congress
  • Place of Presentation: Pacifico Yokohama (神奈川県）