Fig. 1.

Distribution of H. pylori in gnotobiotic tox176 mice. (A and B) Multilabel immunohistochemical study of stomachs from 7-week-old germ-free normal (A) and tox176 (B) mice showing that tox176-mediated parietal cell ablation results in amplification of mitotically active GEPs. Animals were treated with BrdUrd (green) 90 min before death to label cells in S-phase. Dolichos biflorus agglutinin (red) is used to mark parietal cells. Pit cells are tagged with Alexa Fluor 350-conjugated Anguilla anguilla agglutinin (blue), and neck cells are tagged with Alexa Fluor 350- and Alexa Fluor 647-tagged Griffonia simplicifolia II lectin (purple). GEPs produce NeuAcα2,3Galβ1,4 glycans (white after staining with biotinylated MAA and Cy3-labeled streptavidin). (C) Attachment of CAG7:8 (red) to NeuAcα2,3Galβ1,4-positive GEPs (marked green with MAA) in a stomach from a 15-week-old gnotobiotic tox176 mouse killed after 4 weeks of infection. Nuclei are stained blue with bis-benzimide. (D) Frame from a 3D confocal microscopic projection of a focal area of infection in a tox176 stomach showing intracellular bacteria (red, arrows). Cell borders are delineated by using an antibody to E-cadherin (green after treatment with Alexa Fluor 488-tagged donkey anti-rat Ig). To view the 3D projection in its entirety, see Movie 1, which is published as supporting information on the PNAS web site. (Bars, 10 μm.)