title: Functional modulation of human intestinal epithelial cell responses by Bifidobacterium infantis and Lactobacillus salivarius.
abstract: Intestinal epithelial cells (IECs) and dendritic cells (DCs) play a pivotal role in antigen sampling and the maintenance of gut homeostasis. However, the interaction of commensal bacteria with the intestinal surface remains incompletely understood. Here we investigated immune cell responses to commensal and pathogenic bacteria. HT-29 human IECs were incubated with Bifidobacterium infantis 35624, Lactobacillus salivarius UCC118 or Salmonella typhimurium UK1 for varying times, or were pretreated with a probiotic for 2 hr prior to stimulation with S. typhimurium or flagellin. Gene arrays were used to examine inflammatory gene expression. Nuclear factor (NF)-kappaB activation, interleukin (IL)-8 secretion, pathogen adherence to IECs, and mucin-3 (MUC3) and E-cadherin gene expression were assayed by TransAM assay, enzyme-linked immunosorbent assay (ELISA), fluorescence, and real-time reverse transcriptase-polymerase chain reaction (RT-PCR), respectively. IL-10 and tumour necrosis factor (TNF)-alpha secretion by bacteria-treated peripheral blood-derived DCs were measured using ELISA. S. typhimurium increased expression of 36 of the 847 immune-related genes assayed, including NF-kappaB and IL-8. The commensal bacteria did not alter expression levels of any of the 847 genes. However, B. infantis and L. salivarius attenuated both IL-8 secretion at baseline and S. typhimurium-induced pro-inflammatory responses. B. infantis also limited flagellin-induced IL-8 protein secretion. The commensal bacteria did not increase MUC3or E-cadherin expression, or interfere with pathogen binding to HT-29 cells, but they did stimulate IL-10 and TNF-alpha secretion by DCs. The data demonstrate that, although the intestinal epithelium is immunologically quiescent when it encounters B. infantis or L. salivarius, these commensal bacteria exert immunomodulatory effects on intestinal immune cells that mediate host responses to flagellin and enteric pathogens.
authors.affiliation: Alimentary Pharmabiotic Centre, University College Cork, National University of Ireland, Cork, Ireland.
authors.forename: Ann M
authors.initials: AM
authors.lastname: O'Hara
authors.forename: Padraig
authors.initials: P
authors.lastname: O'Regan
authors.forename: Aine
authors.initials: A
authors.lastname: Fanning
authors.forename: Caitlin
authors.initials: C
authors.lastname: O'Mahony
authors.forename: John
authors.initials: J
authors.lastname: Macsharry
authors.forename: Anne
authors.initials: A
authors.lastname: Lyons
authors.forename: John
authors.initials: J
authors.lastname: Bienenstock
authors.forename: Liam
authors.initials: L
authors.lastname: O'Mahony
authors.forename: Fergus
authors.initials: F
authors.lastname: Shanahan
mesh_terms: D000942:Antigens, Bacterial / Q000276:immunology; D001644:Bifidobacterium / Q000276:immunology*; D015820:Cadherins / Q000096:biosynthesis / Q000235:genetics; D002470:Cell Survival / Q000276:immunology; D003106:Colon / Q000276:immunology*; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000276:immunology; D004847:Epithelial Cells / Q000276:immunology; D005408:Flagellin / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology; D019073:HT29 Cells; D006801:Humans; D018928:Immunity, Mucosal; D018836:Inflammation Mediators / Q000276:immunology; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics; D007413:Intestinal Mucosa / Q000276:immunology*; D007778:Lactobacillus / Q000276:immunology*; D055263:Mucin-3; D009077:Mucins / Q000096:biosynthesis / Q000235:genetics; D016328:NF-kappa B / Q000378:metabolism; D019936:Probiotics / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012486:Salmonella typhimurium / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D015820:Cadherins; D016207:Cytokines; D018836:Inflammation Mediators; D016209:Interleukin-8; D055263:Mucin-3; D009077:Mucins; D016328:NF-kappa B; D012333:RNA, Messenger; D005408:Flagellin