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In Vitro Evidence for Immune-Modulatory Properties of Non-Digestible Oligosaccharides: Direct Effect on Human Monocyte Derived Dendritic Cells.

Lehmann S, Hiller J, van Bergenhenegouwen J, Knippels LM, Garssen J, Traidl-Hoffmann C - PLoS ONE (2015)

Bottom Line: Oligosaccharides did not induce relevant amounts of IL-12p70 production, but did promote IL-10 release by MoDC.Furthermore, scGOS/lcFOS mixtures exerted a significant enhancing effect on LAB induced IL-10 secretion by MoDC while no increase in IL-12p70 production was observed.Our results indicate anti-inflammatory and direct, microbiota independent, immune-modulatory properties of scGOS/lcFOS mixtures on human MoDC suggesting a possible induction of regulatory T cells (Tregs).

View Article: PubMed Central - PubMed

Affiliation: Institute of Environmental Medicine, UNIKA-T, Technische Universität München, Augsburg, Germany; Center of Allergy and Environment (ZAUM), Member of the German Center for Lung Research (DZL), Technische Universität & Helmholtz Zentrum München, Munich, Germany.

ABSTRACT
Infant formulas containing non-digestible oligosaccharides (NDO) similar to the composition in breast milk or a combination of lactic acid bacteria (LAB) and NDO have been shown to harbor preventive effects towards immune-regulatory disorders. The aim of this study was to investigate the immune-modulatory potential of non-digestible short chain galacto- and long chain fructo-oligosaccharides (scGOS/lcFOS) mimicking the natural distribution of oligosaccharides in human breast milk in presence or absence of certain LAB strains in human monocyte derived dendritic cells (MoDC). Immature human MoDC prepared from peripheral blood of healthy non-atopic volunteers were screened in vitro after stimulation with specific scGOS/lcFOS in presence or absence of LAB. IL-10 and IL-12p70 release was analyzed after 24 hours in cell-free supernatants by enzyme-linked immunosorbent assay (ELISA). A luminex-based assay was conducted to assess further cytokine and chemokine release by MoDC. To investigate the resulting T cell response, stimulated MoDC were co-incubated with naïve T cells in allogeneic stimulation assays and intracellular Foxp3 expression, as well as immune-suppressive capacity was determined. Oligosaccharides did not induce relevant amounts of IL-12p70 production, but did promote IL-10 release by MoDC. Furthermore, scGOS/lcFOS mixtures exerted a significant enhancing effect on LAB induced IL-10 secretion by MoDC while no increase in IL-12p70 production was observed. Blocking toll like receptor (TLR)4 abrogated the increase in IL-10 in both the direct stimulation and the LAB stimulation of MoDC, suggesting that scGOS/lcFOS act via TLR4. Finally, scGOS/lcFOS-treated MoDC were shown to upregulate the number of functional suppressive Foxp3 positive T cells following allogeneic stimulation. Our results indicate anti-inflammatory and direct, microbiota independent, immune-modulatory properties of scGOS/lcFOS mixtures on human MoDC suggesting a possible induction of regulatory T cells (Tregs). The tested combinations of LAB and scGOS/lcFOS might represent a useful dietary ingredient for the maintenance of intestinal homeostasis via the induction of Tregs.

No MeSH data available.


Related in: MedlinePlus

Stimulation with scGOS/lcFOS directly induces a dose dependent enhanced IL-10 release by LAB-primed MoDC that is impaired by addition of a TLR4 antagonist.Immature human MoDC were exposed to oligosaccharide mixture scGOS/lcFOS (312 μg/ml–5 mg/ml) in the presence of B.breve at a concentration of 1x106cfu/ml in the presence (black marks) or absence (grey marks) of a TLR4 antagonist. The amount of IL-10 was measured by ELISA in cell-free supernatant after 24h. Results are presented as mean ± SEM, five independent experiments are shown. * p<0.05, Mann Whitney test. scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharide-mixture, MoDC = monocyte-derived dendritic cells, TLR = Toll like receptor.
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pone.0132304.g005: Stimulation with scGOS/lcFOS directly induces a dose dependent enhanced IL-10 release by LAB-primed MoDC that is impaired by addition of a TLR4 antagonist.Immature human MoDC were exposed to oligosaccharide mixture scGOS/lcFOS (312 μg/ml–5 mg/ml) in the presence of B.breve at a concentration of 1x106cfu/ml in the presence (black marks) or absence (grey marks) of a TLR4 antagonist. The amount of IL-10 was measured by ELISA in cell-free supernatant after 24h. Results are presented as mean ± SEM, five independent experiments are shown. * p<0.05, Mann Whitney test. scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharide-mixture, MoDC = monocyte-derived dendritic cells, TLR = Toll like receptor.

Mentions: To investigate this differential effect on cytokine release in more detail, a larger selection of MoDC-derived soluble mediators was analyzed. scGOS/lcFOS was found to enhance mediator release depending both on the analyte measured and the bacterial strain used suggesting strain-dependent effects (S5 Fig). To investigate the potential contribution of TLR4 activity to the observed effect, MoDC were stimulated in the presence of a LPS antagonist. scGOS/lcFOS dose-dependently increased B.breve-induced MoDC IL-10 release. Blocking of TLR4-activity abrogated this enhanced effect (Fig 5).


In Vitro Evidence for Immune-Modulatory Properties of Non-Digestible Oligosaccharides: Direct Effect on Human Monocyte Derived Dendritic Cells.

Lehmann S, Hiller J, van Bergenhenegouwen J, Knippels LM, Garssen J, Traidl-Hoffmann C - PLoS ONE (2015)

Stimulation with scGOS/lcFOS directly induces a dose dependent enhanced IL-10 release by LAB-primed MoDC that is impaired by addition of a TLR4 antagonist.Immature human MoDC were exposed to oligosaccharide mixture scGOS/lcFOS (312 μg/ml–5 mg/ml) in the presence of B.breve at a concentration of 1x106cfu/ml in the presence (black marks) or absence (grey marks) of a TLR4 antagonist. The amount of IL-10 was measured by ELISA in cell-free supernatant after 24h. Results are presented as mean ± SEM, five independent experiments are shown. * p<0.05, Mann Whitney test. scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharide-mixture, MoDC = monocyte-derived dendritic cells, TLR = Toll like receptor.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493044&req=5

pone.0132304.g005: Stimulation with scGOS/lcFOS directly induces a dose dependent enhanced IL-10 release by LAB-primed MoDC that is impaired by addition of a TLR4 antagonist.Immature human MoDC were exposed to oligosaccharide mixture scGOS/lcFOS (312 μg/ml–5 mg/ml) in the presence of B.breve at a concentration of 1x106cfu/ml in the presence (black marks) or absence (grey marks) of a TLR4 antagonist. The amount of IL-10 was measured by ELISA in cell-free supernatant after 24h. Results are presented as mean ± SEM, five independent experiments are shown. * p<0.05, Mann Whitney test. scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharide-mixture, MoDC = monocyte-derived dendritic cells, TLR = Toll like receptor.
Mentions: To investigate this differential effect on cytokine release in more detail, a larger selection of MoDC-derived soluble mediators was analyzed. scGOS/lcFOS was found to enhance mediator release depending both on the analyte measured and the bacterial strain used suggesting strain-dependent effects (S5 Fig). To investigate the potential contribution of TLR4 activity to the observed effect, MoDC were stimulated in the presence of a LPS antagonist. scGOS/lcFOS dose-dependently increased B.breve-induced MoDC IL-10 release. Blocking of TLR4-activity abrogated this enhanced effect (Fig 5).

Bottom Line: Oligosaccharides did not induce relevant amounts of IL-12p70 production, but did promote IL-10 release by MoDC.Furthermore, scGOS/lcFOS mixtures exerted a significant enhancing effect on LAB induced IL-10 secretion by MoDC while no increase in IL-12p70 production was observed.Our results indicate anti-inflammatory and direct, microbiota independent, immune-modulatory properties of scGOS/lcFOS mixtures on human MoDC suggesting a possible induction of regulatory T cells (Tregs).

View Article: PubMed Central - PubMed

Affiliation: Institute of Environmental Medicine, UNIKA-T, Technische Universität München, Augsburg, Germany; Center of Allergy and Environment (ZAUM), Member of the German Center for Lung Research (DZL), Technische Universität & Helmholtz Zentrum München, Munich, Germany.

ABSTRACT
Infant formulas containing non-digestible oligosaccharides (NDO) similar to the composition in breast milk or a combination of lactic acid bacteria (LAB) and NDO have been shown to harbor preventive effects towards immune-regulatory disorders. The aim of this study was to investigate the immune-modulatory potential of non-digestible short chain galacto- and long chain fructo-oligosaccharides (scGOS/lcFOS) mimicking the natural distribution of oligosaccharides in human breast milk in presence or absence of certain LAB strains in human monocyte derived dendritic cells (MoDC). Immature human MoDC prepared from peripheral blood of healthy non-atopic volunteers were screened in vitro after stimulation with specific scGOS/lcFOS in presence or absence of LAB. IL-10 and IL-12p70 release was analyzed after 24 hours in cell-free supernatants by enzyme-linked immunosorbent assay (ELISA). A luminex-based assay was conducted to assess further cytokine and chemokine release by MoDC. To investigate the resulting T cell response, stimulated MoDC were co-incubated with naïve T cells in allogeneic stimulation assays and intracellular Foxp3 expression, as well as immune-suppressive capacity was determined. Oligosaccharides did not induce relevant amounts of IL-12p70 production, but did promote IL-10 release by MoDC. Furthermore, scGOS/lcFOS mixtures exerted a significant enhancing effect on LAB induced IL-10 secretion by MoDC while no increase in IL-12p70 production was observed. Blocking toll like receptor (TLR)4 abrogated the increase in IL-10 in both the direct stimulation and the LAB stimulation of MoDC, suggesting that scGOS/lcFOS act via TLR4. Finally, scGOS/lcFOS-treated MoDC were shown to upregulate the number of functional suppressive Foxp3 positive T cells following allogeneic stimulation. Our results indicate anti-inflammatory and direct, microbiota independent, immune-modulatory properties of scGOS/lcFOS mixtures on human MoDC suggesting a possible induction of regulatory T cells (Tregs). The tested combinations of LAB and scGOS/lcFOS might represent a useful dietary ingredient for the maintenance of intestinal homeostasis via the induction of Tregs.

No MeSH data available.


Related in: MedlinePlus