Multiplex-PCR-Based Screening and Computational Modeling of Virulence Factors and T-Cell Mediated Immunity in Helicobacter pylori Infections for Accurate Clinical Diagnosis

PLoS One. 2015 Aug 19;10(8):e0136212. doi: 10.1371/journal.pone.0136212. eCollection 2015.

Abstract

The outcome of H. pylori infection is closely related with bacteria's virulence factors and host immune response. The association between T cells and H. pylori infection has been identified, but the effects of the nine major H. pylori specific virulence factors; cagA, vacA, oipA, babA, hpaA, napA, dupA, ureA, ureB on T cell response in H. pylori infected patients have not been fully elucidated. We developed a multiplex- PCR assay to detect nine H. pylori virulence genes with in a three PCR reactions. Also, the expression levels of Th1, Th17 and Treg cell specific cytokines and transcription factors were detected by using qRT-PCR assays. Furthermore, a novel expert derived model is developed to identify set of factors and rules that can distinguish the ulcer patients from gastritis patients. Within all virulence factors that we tested, we identified a correlation between the presence of napA virulence gene and ulcer disease as a first data. Additionally, a positive correlation between the H. pylori dupA virulence factor and IFN-γ, and H. pylori babA virulence factor and IL-17 was detected in gastritis and ulcer patients respectively. By using computer-based models, clinical outcomes of a patients infected with H. pylori can be predicted by screening the patient's H. pylori vacA m1/m2, ureA and cagA status and IFN-γ (Th1), IL-17 (Th17), and FOXP3 (Treg) expression levels. Herein, we report, for the first time, the relationship between H. pylori virulence factors and host immune responses for diagnostic prediction of gastric diseases using computer-based models.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Computer Simulation
  • Diagnosis, Computer-Assisted
  • Expert Systems
  • Gastritis / diagnosis
  • Gastritis / immunology
  • Gastritis / microbiology
  • Genes, Bacterial
  • Helicobacter Infections / diagnosis*
  • Helicobacter Infections / immunology*
  • Helicobacter Infections / microbiology
  • Helicobacter pylori* / genetics
  • Helicobacter pylori* / immunology
  • Helicobacter pylori* / pathogenicity
  • Host-Pathogen Interactions / genetics
  • Host-Pathogen Interactions / immunology
  • Humans
  • Immunity, Cellular
  • Multiplex Polymerase Chain Reaction
  • T-Lymphocyte Subsets / immunology
  • Virulence Factors / genetics

Substances

  • Bacterial Proteins
  • Virulence Factors

Grants and funding

The financial support of this work was provided by Istanbul Technical University through Scientific Research Project Funds with the grant number 36243.