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A Novel Gastric Spheroid Co-Culture Model Reveals Chemokine-Dependent Recruitment of Human Dendritic Cells to the Gastric Epithelium.
Cellular and Molecular Gastroenterology and Hepatology 2019 March 14
BACKGROUND & AIMS: Gastric dendritic cells (DCs) control the adaptive response to infection with Helicobacter pylori, a major risk factor for peptic ulcer disease and gastric cancer. We hypothesize that DC interactions with the gastric epithelium position gastric DCs for uptake of luminal H. pylori and promote DC responses to epithelial-derived mediators. The aim of this study was to determine whether the gastric epithelium actively recruits DCs, using a novel co-culture model of human gastric epithelial spheroids and monocyte-derived DCs.
METHODS: Spheroid cultures of primary gastric epithelial cells were infected with H. pylori by microinjection. Co cultures were established by adding human monocyte-derived DCs (MoDCs) to the spheroid cultures and were analyzed for DC recruitment and antigen uptake by confocal microscopy. Protein array, gene expression PCR array and chemotaxis assays were used to identify epithelial-derived chemotactic factors that attract DCs. Data from the co-culture model were confirmed using human gastric tissue samples.
RESULTS: Human MoDCs co-cultured with gastric spheroids spontaneously migrated to the gastric epithelium, established tight interactions with the epithelial cells and phagocytosed luminally applied H. pylori. DC recruitment was increased upon H. pylori infection of the spheroids and involved the activity of multiple chemokines including CXCL1, CXCL16, CXCL17 and CCL20. Enhanced chemokine expression and DC recruitment to the gastric epithelium was also observed in H. pylori-infected human gastric tissue samples.
CONCLUSIONS: Our results indicate that the gastric epithelium actively recruits DCs for immunosurveillance and pathogen sampling through chemokine-dependent mechanisms, with increased recruitment upon active H. pylori infection.
METHODS: Spheroid cultures of primary gastric epithelial cells were infected with H. pylori by microinjection. Co cultures were established by adding human monocyte-derived DCs (MoDCs) to the spheroid cultures and were analyzed for DC recruitment and antigen uptake by confocal microscopy. Protein array, gene expression PCR array and chemotaxis assays were used to identify epithelial-derived chemotactic factors that attract DCs. Data from the co-culture model were confirmed using human gastric tissue samples.
RESULTS: Human MoDCs co-cultured with gastric spheroids spontaneously migrated to the gastric epithelium, established tight interactions with the epithelial cells and phagocytosed luminally applied H. pylori. DC recruitment was increased upon H. pylori infection of the spheroids and involved the activity of multiple chemokines including CXCL1, CXCL16, CXCL17 and CCL20. Enhanced chemokine expression and DC recruitment to the gastric epithelium was also observed in H. pylori-infected human gastric tissue samples.
CONCLUSIONS: Our results indicate that the gastric epithelium actively recruits DCs for immunosurveillance and pathogen sampling through chemokine-dependent mechanisms, with increased recruitment upon active H. pylori infection.
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