Abstract
Helicobacter pylori colonises the gastric mucosa of humans. The majority of organisms live in mucus. These organisms are an important reservoir for infection of the underlying epithelium. Cell culture models for H. pylori infection do not normally possess a mucus layer. The interaction of H. pylori with TFF1, a member of the trefoil factor family found in gastric mucin, is mediated by lipopolysaccharide. To test the hypothesis that the interaction of H. pylori with TFF1 promotes mucus colonization we characterised the interaction of H. pylori with a mucus secreting cell line, HT29-MTX-E12. An isogenic mutant of H. pylori with truncated core oligosaccharides was produced and binding to TFF1 and ability to colonise HT29-MTX-E12 cells determined. The adherent mucus layer of HT29-MTX-E12 cells contained the gastric mucin MUC5AC and trefoil factors, TFF1 and TFF3. H. pylori was found within the mucus layer in discrete clusters and in close association with TFF1. It also interacted with the membrane bound mucin MUC1 and replicated when co-cultured with the cells. An isogenic mutant of H. pylori with a truncated LPS core did not interact with TFF1, and colonization of HT29-MTX-E12 cells was reduced compared to the wild-type strain (p<0.05). Preincubation of cells with wild type LPS but not with truncated LPS resulted in reduced colonization by H. pylori. These results demonstrate that the interaction of TFF1 with H. pylori is important for colonization of gastric mucus and the core oligosaccharide of H. pylori LPS is critical for this interaction to occur. HT29-MTX-E12 cells are a useful system with which to study the interaction of bacteria with mucosal surfaces and the effect of such interactions on mediating colonization.
Highlights
The majority of bacterial infections in humans and animals result from pathogens colonizing the body via mucosal surfaces such as the gastrointestinal, respiratory and urinary tracts
We have previously identified low molecular weight or rough form (RF) lipopolysacharide (LPS) of H. pylori as a bacterial factor that interacts with TFF1 [9]
The aim of the present study was to test the hypothesis that the interaction of H. pylori with TFF1 promotes colonization of gastric mucus and that the core oligosaccharide of H. pylori LPS is the critical bacterial factor that mediates the interaction between H. pylori and TFF1
Summary
The majority of bacterial infections in humans and animals result from pathogens colonizing the body via mucosal surfaces such as the gastrointestinal, respiratory and urinary tracts. Helicobacter pylori colonizes the gastric mucosa of humans and primates. H. pylori has been described as a paradigm for chronic infection of mucosal surfaces [2]. The majority of infecting bacteria live in the mucus layer that overlies the gastric epithelial cells [3] and colonization of experimental animals suggests that the organisms live close to the epithelial surface [4]. Elucidation of the mechanisms that H. pylori uses to colonise mucosal surfaces could give us valuable insight into how pathogens overcome the barriers to infection such as the presence of a mucus layer
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