title: Differential effect of immune cells on non-pathogenic Gram-negative bacteria-induced nuclear factor-kappaB activation and pro-inflammatory gene expression in intestinal epithelial cells.
abstract: We have previously shown that non-pathogenic Gram negative bacteria induce RelA phosphorylation, nuclear factor (NF)-kappaB transcriptional activity and pro-inflammatory gene expression in intestinal epithelial cells (IEC) in vivo and in vitro. In this study, we investigated the molecular mechanism of immune-epithelial cell cross-talk on Gram-negative enteric bacteria-induced NF-kappaB signalling and pro-inflammatory gene expression in IEC using HT-29/MTX as well as CaCO-2 transwell cultures Interestingly, while differentiated HT-29/MTX cells are unresponsive to non-pathogenic Gram negative bacterial stimulation, interleukin-8 (IL-8) mRNA accumulation is strongly induced in Escherichia coli- but not Bacteroides vulgatus-stimulated IEC cocultured with peripheral blood (PBMC) and lamina propria mononuclear cells (LPMC). The presence of PBMC triggered both E. coli- and B. vulgatus-induced mRNA expression of the Toll-like receptor-4 accessory protein MD-2 as well as endogenous IkappaBalpha phosphorylation, demonstrating similar capabilities of these bacteria to induce proximal NF-kappaB signalling. However, B. vulgatus failed to trigger IkappaBalpha degradation and NF-kappaB transcriptional activity in the presence of PBMC. Interestingly, B. vulgatus- and E. coli-derived lipopolysaccharide-induced similar IL-8 mRNA expression in epithelial cells after basolateral stimulation of HT-29/PBMC cocultures. Although luminal enteric bacteria have adjuvant and antigenic properties in chronic intestinal inflammation, PBMC from patients with active ulcerative colitis and Crohn's disease differentially trigger epithelial cell activation in response to E. coli and E. coli-derived LPS. In conclusion, this study provides evidence for a differential regulation of non-pathogenic Gram-negative bacteria-induced NF-kappaB signalling and IL-8 gene expression in IEC cocultured with immune cells and suggests the presence of mechanisms that assure hyporesponsiveness of the intestinal epithelium to certain commensally enteric bacteria.
authors.affiliation: Department of Medicine and the Center for Gastrointestinal Biology and Disease, University of North Carolina, Chapel Hill, USA. haller@wzw.tum.de
authors.forename: D
authors.initials: D
authors.lastname: Haller
authors.forename: L
authors.initials: L
authors.lastname: Holt
authors.forename: A
authors.initials: A
authors.lastname: Parlesak
authors.forename: J
authors.initials: J
authors.lastname: Zanga
authors.forename: A
authors.initials: A
authors.lastname: Bäuerlein
authors.forename: R B
authors.initials: RB
authors.lastname: Sartor
authors.forename: C
authors.initials: C
authors.lastname: Jobin
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology; D001439:Bacteroides / Q000276:immunology; D018938:Caco-2 Cells; D002478:Cells, Cultured; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000276:immunology; D004926:Escherichia coli / Q000276:immunology; D005786:Gene Expression Regulation / Q000276:immunology; D006090:Gram-Negative Bacteria / Q000276:immunology*; D006801:Humans; D018928:Immunity, Mucosal; D015212:Inflammatory Bowel Diseases / Q000276:immunology* / Q000382:microbiology; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics; D007413:Intestinal Mucosa / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D016328:NF-kappa B / Q000378:metabolism*; D010766:Phosphorylation; D012333:RNA, Messenger / Q000235:genetics; D015398:Signal Transduction / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D016209:Interleukin-8; D016328:NF-kappa B; D012333:RNA, Messenger