Changes of tight junction and interleukin‐8 expression using a human gastroid monolayer model of Helicobacter pylori infection

Background Lack of a model that mirrors Helicobacter pylori‐induced gastric mucosal inflammation has hampered investigation of early host‐bacterial interactions. We used an ex vivo model of human stomach, gastric epithelial organoid monolayers (gastroid monolayers) to investigate interactions of H pylori infection and the apical junctional complex and interleukin‐8 (IL‐8) expression. Method Morphology of human antral mucosal gastroid monolayers was evaluated using histology, immunohistochemical (IHC) staining, and transmission electron microscopy (TEM). Functional and gross changes in the apical junctional complexes were assessed using transepithelial electrical resistance (TEER), cytotoxicity assays, and confocal laser scanning microscopy. IL‐8 expression was evaluated by real‐time quantitative PCR and ELISA. Results When evaluated by IHC and TEM, the morphology of gastroid monolayers closely resembled in vivo human stomach. Following inoculation of H pylori, TEER transiently declined (up to 51%) in an H pylori density‐dependent manner. TEER recovered by 48 hours post‐infection and remained normal despite continued presence and replication of H pylori. Confocal scanning microscopy showed minimal disruption of zonula occludens‐1 or E‐cadherin structure. IL‐8 production was unchanged by infection with either CagA‐positive or CagA‐negative H pylori and JNK and MEK inhibitors did not suppress IL‐8 production, whereas p38 and IKK inhibitor significantly did. Conclusion Human gastroid monolayers provide a model for experimental H pylori infection more consistent with in vivo human infections than seen with typical gastric epithelial cell lines. This ex vivo system should lead to better understanding of H pylori host‐pathogen interactions.