title: Cell-free culture supernatant of Bifidobacterium breve CNCM I-4035 decreases pro-inflammatory cytokines in human dendritic cells challenged with Salmonella typhi through TLR activation.
abstract: Dendritic cells (DCs) constitute the first point of contact between gut commensals and our immune system. Despite growing evidence of the immunomodulatory effects of probiotics, the interactions between the cells of the intestinal immune system and bacteria remain largely unknown. Indeed,, the aim of this work was to determine whether the probiotic Bifidobacterium breve CNCM I-4035 and its cell-free culture supernatant (CFS) have immunomodulatory effects in human intestinal-like dendritic cells (DCs) and how they respond to the pathogenic bacterium Salmonella enterica serovar Typhi, and also to elucidate the molecular mechanisms involved in these interactions. Human DCs were directly challenged with B. breve/CFS, S. typhi or a combination of these stimuli for 4 h. The expression pattern of genes involved in Toll-like receptor (TLR) signaling pathway and cytokine secretion was analyzed. CFS decreased pro-inflammatory cytokines and chemokines in human intestinal DCs challenged with S. typhi. In contrast, the B. breve CNCM I-4035 probiotic strain was a potent inducer of the pro-inflammatory cytokines and chemokines tested, i.e., TNF-α, IL-8 and RANTES, as well as anti-inflammatory cytokines including IL-10. CFS restored TGF-β levels in the presence of Salmonella. Live B.breve and its supernatant enhanced innate immune responses by the activation of TLR signaling pathway. These treatments upregulated TLR9 gene transcription. In addition, CFS was a more potent inducer of TLR9 expression than the probiotic bacteria in the presence of S. typhi. Expression levels of CASP8 and IRAK4 were also increased by CFS, and both treatments induced TOLLIP gene expression. Our results indicate that the probiotic strain B. breve CNCM I-4035 affects the intestinal immune response, whereas its supernatant exerts anti-inflammatory effects mediated by DCs. This supernatant may protect immune system from highly infectious agents such as Salmonella typhi and can down-regulate pro-inflammatory pathways.
authors.affiliation: Department of Biochemistry and Molecular Biology II, Institute of Nutrition and Food Technology José Mataix, Biomedical Research Center, University of Granada, Granada, Spain.
authors.forename: Miriam
authors.initials: M
authors.lastname: Bermudez-Brito
authors.forename: Sergio
authors.initials: S
authors.lastname: Muñoz-Quezada
authors.forename: Carolina
authors.initials: C
authors.lastname: Gomez-Llorente
authors.forename: Esther
authors.initials: E
authors.lastname: Matencio
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Bernal
authors.forename: Fernando
authors.initials: F
authors.lastname: Romero
authors.forename: Angel
authors.initials: A
authors.lastname: Gil
mesh_terms: D001644:Bifidobacterium / Q000502:physiology*; D053181:Caspase 8 / Q000235:genetics / Q000276:immunology; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D016207:Cytokines / Q000235:genetics / Q000276:immunology*; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology; D006801:Humans; D053592:Interleukin-1 Receptor-Associated Kinases / Q000235:genetics / Q000276:immunology; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000276:immunology; D019936:Probiotics / Q000494:pharmacology*; D012485:Salmonella typhi / Q000254:growth & development / Q000276:immunology*; D015398:Signal Transduction; D051217:Toll-Like Receptor 9 / Q000235:genetics / Q000276:immunology
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D047908:Intracellular Signaling Peptides and Proteins; C495371:TLR9 protein, human; C410602:TOLLIP protein, human; D051217:Toll-Like Receptor 9; C507773:IRAK4 protein, human; D053592:Interleukin-1 Receptor-Associated Kinases; C505530:CASP8 protein, human; D053181:Caspase 8
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