Fig. 2From: Upregulation of MUC5AC production and deposition of LEWIS determinants by HELICOBACTER PYLORI facilitate gastric tissue colonization and the maintenance of infectionAdhesion of H. pylori to guinea pig gastric epithelial cells mediated by MUC5AC mucin and Lewis (Le)X/LeY determinants. Binding of H. pylori to guinea pig primary gastric epithelial cells and gastric tissue specimens was evaluated by imaging H. pylori stained with anti-H. pylori antibodies (Ab) conjugated with fluorescein isothiocyanate (FITC) in a fluorescence reader or confocal microscope, counterstained with DAPI or phalloidine. a – guinea pig primary gastric epithelial cells non treated (in culture medium alone) or treated with selected H. pylori components: for 24 h with glycine acid extract – GE (10 μg/ml), urease subunite A – UreA (5 μg/ml), cytotoxin associated gene A (CagA) protein (1 μg/ml), H. pylori lipopolysaccharide (LPS Hp) or E.coli LPS –LPS Ec (25 ng/ml) or for 2 h with live H. pylori – Hp (2 × 107 colony forming units - CFU/ml) were prepared. Further, cells non-treated (control of adhesion) or blocked with anti-MUC5AC or with anti-LeX/LeY antibodies were used in adhesion assay. a i – the intensity of fluorescence measured in a fluorescence reader at 495 nm (excitation) and 519 nm (emission), mean values ±SD. * Statistical significance for cells treated with an individual component vs untreated cells or # treated with individual antibody vs untreated cells (control cells), p < 0.05 in non parametric U Mann-Whitney test. Representative images of primary gastric epithelial cells untreated (a ii) or treated (a iii) with anti-MUC5AC antibodies before H. pylori binding experiments, stained with anti-H. pylori FITC antibodies in confocal microscope (Leica TCS SPE), at 495 nm excitation and 519 emission for FITC, 345 excitation/455 emission for DAPI and 591 nm excitation/608 nm emission for phalloidine (640 magnification). Gastric tissue specimens from non infected guinea pigs untreated or treated with anti-MUC5AC (b) or anti-LeX (c) blocking antibodies before 2 h exposure to live H. pylori (2 × 107 colony forming units (CFU)/ml). bi and Ci – intensity of fluorescence measured using the software ImageJ version 1.48v (National Institute of Health, United States) at 495 excitation/519 emission, mean values ±SD. *Statistically significant values for gastric tissue blocked with anti-MUC5AC or anti-LeX antibodies vs unblocked specimens. Representative images of H. pylori adhesion to the guinea pig gastric tissue non treated with anti-MUC5AC (bii) or anti-LeX (cii) antibodies vs gastric tissue treated with such antibodies (b iii, iii) from fluorescence microscope (Axio Scope A1, Zeiss, Germany), at 495 nm excitation/519 nm emission for FITC, 345 nm excitation/455 nm emission for DAPI and 591 nm excitation/608 nm emission for phalloidine (100 × magnification)Back to article page