Abstract
BackgroundHelicobacter pylori bacteria colonize human gastric mucosa, cause chronic inflammation, peptic ulcers and gastric cancer. Colonization is mediated by H. pylori adhesins, which preferentially bind mucin 5 (MUC5AC) and Lewis (Le) determinants. The aim of this study was to evaluate the influence of H. pylori and their components on MUC5AC production and deposition of LeX/LeY in gastric epithelial cells in relation to bacterial adhesion using Caviae porcellus primary gastric epithelial cells and an in vivo model of experimental H. pylori infection in these animals.MethodsMUCA5C and LeX/LeY were induced in vitro by live H. pylori reference strain CCUG 17874 (2 × 107 CFU/ml), H. pylori glycine acid extract (GE), 10 μg/ml; cytotoxin associated gene A (CagA) protein, 1 μl/ml; UreA urease subunit, 5 μg/ml; lipopolysaccharide (LPS) 25 ng/ml and imaged by fluorescence microscopy after anti-MUC5AC or anti-LeX/LeY FITC antibody staining. Bacterial adhesion was imaged by using anti-H. pylori FITC antibodies. The animals were inoculated per os with H. pylori (3 times in 2 days intervals, 1 × 1010 CFU/ml). After 7 or 28 days an infection and inflammation were assessed by histological, serological and molecular methods. Gastric tissue sections of infected and control animals were screend for MUCA5C and LeX, and H. pylori adhesion as above.ResultsMUC5AC production and deposition of Lewis determinants, especially LeX were upregulated in the milieu of live H. pylori as well as GE, CagA, UreA or LPS in vitro and in vivo during infection, more effectively in the acute (7 days) than in the chronic (28 days) phase of infection. This was related to enhanced adhesion of H. pylori, which was abrogated by anti-MUC5AC and anti-LeX or anti-LeY antibody treatment.ConclusionsModulation of MUCA5C production and LeX/LeY deposition in the gastric mucosa by H. pylori can significantly increase gastric tissue colonization during H. pylori infection.
Highlights
The gastric mucosa is a physical barrier covered with a mucus layer, which protects stomach against harmful chemical, enzymatic, microbiological and mechanical factors [1, 2]
mucin 5 (MUC5AC) and LeX/LeY production by Guinea pigs gastric epithelial cells in the milieu of live H. pylori or soluble compounds of these bacteria in relation to H. pylori adhesion – In vitro and in vivo models We were focused on answering the question whether live H. pylori rods or their soluble antigens modulate MUC5AC production by the gastric epithelial cells and LeX/LeY deposition, and how it influences the process of colonization of gastric mucosa by H. pylori using an in vitro model of primary gastric epithelial cells derived from the guinea pig gastric tissue
Production of MUC5AC was significantly increased after 24 h stimulation of guinea pig primary gastric epithelial cells with H. pylori surface components glycine acid extract (GE) (10 μg/ml), cytotoxin associated gene A (CagA) (1 μg/ml), UreA (5 μg/ml), H. pylori LPS (25 ng/ ml) as well as after 2 h stimulation with live H. pylori (2 × 107 colony forming unit (CFU)/ml) (Fig. 1a), p < 0.05 in Kruskal-Wallis test
Summary
The gastric mucosa is a physical barrier covered with a mucus layer, which protects stomach against harmful chemical, enzymatic, microbiological and mechanical factors [1, 2]. By using two Caviae porcellus (guinea pig) models: a model of primary gastric epithelial cells and a model of experimental H. pylori infection, we focused on the relation between MUC5AC and LeX/LeY production in response to gastric epithelial cell exposure in vivo or in vitro to H. pylori reference LeX/Y positive strain or soluble components of these bacteria, and the effectiveness of epithelial cell colonization. We considerd both the host and bacterial LeX/LeY components in the course of H. pylori adhesion to gastric epithelial cells on guinea pig model. The aim of this study was to evaluate the influence of H. pylori and their components on MUC5AC production and deposition of LeX/LeY in gastric epithelial cells in relation to bacterial adhesion using Caviae porcellus primary gastric epithelial cells and an in vivo model of experimental H. pylori infection in these animals
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