Proteomic characterization of Helicobacter pylori CagA antigen recognized by child serum antibodies and its epitope mapping by peptide array

PLoS One. 2014 Aug 20;9(8):e104611. doi: 10.1371/journal.pone.0104611. eCollection 2014.

Abstract

Serum antibodies against pathogenic bacteria play immunologically protective roles, and can be utilized as diagnostic markers of infection. This study focused on Japanese child serum antibodies against Helicobacter pylori, a chronically-infected gastric bacterium which causes gastric cancer in adults. Serological diagnosis for H. pylori infection is well established for adults, but it needs to be improved for children. 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 protein (CagA) 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, modified from our previous protein chip method. Each of the epitopes was found to contain amino acid residue(s) unique to East Asian CagA. Epitope analysis of CagA indicated importance of the regional CagA antigens for serodiagnosis of H. pylori infection in children.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Bacterial / immunology*
  • Bacterial Proteins / immunology*
  • Child
  • Child, Preschool
  • Epitope Mapping
  • Helicobacter Infections / diagnosis*
  • Helicobacter Infections / immunology
  • Helicobacter pylori / immunology*
  • Humans
  • Proteomics

Substances

  • Antigens, Bacterial
  • Bacterial Proteins
  • cagA protein, Helicobacter pylori

Grants and funding

This work was supported by KAKEN-HI No. 24590919 from JSPS of Japan, by Yamaguchi University Innovation Center Foundation grant No. 2010-03, and partially by Yamaguchi University Yobimizu Project 2013, to JA. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of manuscript.