Takahiro Uotani1,2, Kosuke Murakami3, Tomohisa Uchida4, Shingo Tanaka1,2, Hiroyuki Nagashima5, Xi-Lei Zeng3, Junko Akada2, Mary K Estes3, David Y Graham1, Yoshio Yamaoka1,2. 1. Department of Gastroenterology and Hepatology, Baylor College of Medicine and Michael E. DeBakey Veterans Affairs Medical Center, Houston, Texas. 2. Department of Environmental and Preventive Medicine, Oita University Faculty of Medicine, Yufu, Japan. 3. Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas. 4. Department of Molecular Pathology, Oita University Faculty of Medicine, Yufu, Japan. 5. Gastroenterological Medicine, Rumoi City Hospital, Rumoi, Japan.
Abstract
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.
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 pyloriinfection 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:Humangastroid monolayers provide a model for experimental H pyloriinfection more consistent with in vivo humaninfections than seen with typical gastric epithelial cell lines. This ex vivo system should lead to better understanding of H pylori host-pathogen interactions.
Authors: John M Russo; Peter Florian; Le Shen; W Vallen Graham; Maria S Tretiakova; Alfred H Gitter; Randall J Mrsny; Jerrold R Turner Journal: Gastroenterology Date: 2005-04 Impact factor: 22.682
Authors: Maria Jose Oliveira; Angela Margarida Costa; Ana Catarina Costa; Rui Manuel Ferreira; Paula Sampaio; Jose Carlos Machado; Raquel Seruca; Marc Mareel; Ceu Figueiredo Journal: J Infect Dis Date: 2009-09-01 Impact factor: 5.226
Authors: Yoshio Yamaoka; Takahiko Kudo; Hong Lu; Antonella Casola; Allan R Brasier; David Y Graham Journal: Gastroenterology Date: 2004-04 Impact factor: 22.682
Authors: Nina Bertaux-Skeirik; Rui Feng; Michael A Schumacher; Jing Li; Maxime M Mahe; Amy C Engevik; Jose E Javier; Richard M Peek; Karen Ottemann; Veronique Orian-Rousseau; Gregory P Boivin; Michael A Helmrath; Yana Zavros Journal: PLoS Pathog Date: 2015-02-06 Impact factor: 6.823
Authors: Amanda D Morgan; Kevin D Seely; Lauren D Hagenstein; Garrett M Florey; James M Small Journal: Cancers (Basel) Date: 2022-10-06 Impact factor: 6.575
Authors: Ju Yup Lee; Nayoung Kim; Yoon Jin Choi; Ji Hyun Park; Hassan Ashktorab; Duane T Smoot; Dong Ho Lee Journal: J Neurogastroenterol Motil Date: 2020-04-30 Impact factor: 4.924