Helicobacter hepaticus cholesterol-α-glucosyltransferase is essential for establishing colonization in male A/JCr mice

Helicobacter. 2014 Aug;19(4):280-8. doi: 10.1111/hel.12135. Epub 2014 May 23.

Abstract

Background: Helicobacter pylori cholesterol-α-glucosyltransferase (cgt) is essential for survival of H. pylori in mice. Enterohepatic H. hepaticus, the cause of colonic and hepatocellular carcinoma in susceptible mouse strains, contains an ortholog of the H. pylori cgt. However, the role of cgt in the pathogenesis of H. hepaticus has not been investigated.

Materials and methods: Two cgt-deficient isogenic mutants of wild-type H. hepaticus (WT) 3B1 were generated and used to inoculate male A/JCr mice. Cecal and hepatic colonization levels of the mutants and WT 3B1 as well as select inflammation-associated cytokines were measured by qPCR at 4 months postinoculation.

Results: Both mutants were undetectable in the cecum of any inoculated mice (10 per mutant) but were detected in two livers (one for each mutant); by contrast, 9 and 7 of 10 mice inoculated with WT 3B1 were qPCR positive in the ceca and livers, respectively. The mice inoculated with the mutants developed significantly less severe hepatic inflammation (p < .05) and also produced significantly lower hepatic mRNA levels of proinflammatory cytokines Ifn-γ (p < .01) and Tnf-α (p ≤ .02) as well as anti-inflammatory factors Il10 and Foxp3 compared with the WT 3B1-inoculated mice. Additionally, the WT 3B1-inoculated mice developed significantly higher Th1-associated IgG2a (p < .0001) and Th2-associated IgG1 responses (p < .0001) to H. hepaticus infection than mice dosed with isogenic cgt mutants.

Conclusion: Our data indicate that the cholesterol-α-glucosyltransferase is required for establishing colonization of the intestine and liver and therefore plays a critical role in the pathogenesis of H. hepaticus.

Keywords: Helicobacter hepaticus; bacterial load; cholesterol; model; mouse; virulence genes.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cecum / microbiology
  • Cholesterol / metabolism*
  • Cytokines / biosynthesis
  • Disease Models, Animal
  • Gene Deletion
  • Glucosyltransferases / genetics
  • Glucosyltransferases / metabolism*
  • Helicobacter Infections / microbiology*
  • Helicobacter hepaticus / enzymology*
  • Helicobacter hepaticus / genetics
  • Helicobacter hepaticus / growth & development*
  • Liver / microbiology
  • Male
  • Mice, Inbred A
  • Real-Time Polymerase Chain Reaction
  • Virulence Factors / genetics
  • Virulence Factors / metabolism*

Substances

  • Cytokines
  • Virulence Factors
  • Cholesterol
  • Glucosyltransferases