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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

scGOS/lcFOS stimulated MoDC induce CD4+Foxp3+ T cells.MoDC stimulated with scGOS/lcFOS (5 mg/ml) were used to prime naïve CD4+CD45RA+ T cells in an ASA. At day 7, cells were stained for CD4 and intracellular Foxp3 was analyzed by flow cytometry. The dot plots show data from one representative experiment (A-C). T cells that have been co-incubated with medium-treated MoDC served as medium/negative control (A) and a positive control for Treg polarization is shown in (B). CD4+Foxp3+ T cells induced by scGOS/lcFOS stimulated MoDC are shown in (C). Data in (D) are expressed as percentages of CD4+Foxp3+ cells. Results are presented as mean ± SEM, 4 independent experiments are shown (D). * p<0.05, Mann Whitney test. ASA = allogeneic stimulation assay, scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharides, MoDC = monocyte-derived dendritic cells.
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pone.0132304.g006: scGOS/lcFOS stimulated MoDC induce CD4+Foxp3+ T cells.MoDC stimulated with scGOS/lcFOS (5 mg/ml) were used to prime naïve CD4+CD45RA+ T cells in an ASA. At day 7, cells were stained for CD4 and intracellular Foxp3 was analyzed by flow cytometry. The dot plots show data from one representative experiment (A-C). T cells that have been co-incubated with medium-treated MoDC served as medium/negative control (A) and a positive control for Treg polarization is shown in (B). CD4+Foxp3+ T cells induced by scGOS/lcFOS stimulated MoDC are shown in (C). Data in (D) are expressed as percentages of CD4+Foxp3+ cells. Results are presented as mean ± SEM, 4 independent experiments are shown (D). * p<0.05, Mann Whitney test. ASA = allogeneic stimulation assay, scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharides, MoDC = monocyte-derived dendritic cells.

Mentions: DC release of IL-10 is associated with the induction of Tregs [27]. We therefore investigated whether the IL-10 release following stimulation of MoDC with scGOS/lcFOS would lead to induction of Tregs. MoDC were treated with scGOS/lcFOS for 24 hours, washed and subsequently co-cultured with naïve T cells in an allogeneic stimulation assay. Compared to control MoDC (medium-treated MoDC control Fig 6A, positive Treg control Fig 6B), scGOS/lcFOS-stimulated DC induced an upward shift in Foxp3 expressing T cells (Fig 6C). However, this increase did not reach significance (Fig 6D).


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)

scGOS/lcFOS stimulated MoDC induce CD4+Foxp3+ T cells.MoDC stimulated with scGOS/lcFOS (5 mg/ml) were used to prime naïve CD4+CD45RA+ T cells in an ASA. At day 7, cells were stained for CD4 and intracellular Foxp3 was analyzed by flow cytometry. The dot plots show data from one representative experiment (A-C). T cells that have been co-incubated with medium-treated MoDC served as medium/negative control (A) and a positive control for Treg polarization is shown in (B). CD4+Foxp3+ T cells induced by scGOS/lcFOS stimulated MoDC are shown in (C). Data in (D) are expressed as percentages of CD4+Foxp3+ cells. Results are presented as mean ± SEM, 4 independent experiments are shown (D). * p<0.05, Mann Whitney test. ASA = allogeneic stimulation assay, scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharides, MoDC = monocyte-derived dendritic cells.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0132304.g006: scGOS/lcFOS stimulated MoDC induce CD4+Foxp3+ T cells.MoDC stimulated with scGOS/lcFOS (5 mg/ml) were used to prime naïve CD4+CD45RA+ T cells in an ASA. At day 7, cells were stained for CD4 and intracellular Foxp3 was analyzed by flow cytometry. The dot plots show data from one representative experiment (A-C). T cells that have been co-incubated with medium-treated MoDC served as medium/negative control (A) and a positive control for Treg polarization is shown in (B). CD4+Foxp3+ T cells induced by scGOS/lcFOS stimulated MoDC are shown in (C). Data in (D) are expressed as percentages of CD4+Foxp3+ cells. Results are presented as mean ± SEM, 4 independent experiments are shown (D). * p<0.05, Mann Whitney test. ASA = allogeneic stimulation assay, scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharides, MoDC = monocyte-derived dendritic cells.
Mentions: DC release of IL-10 is associated with the induction of Tregs [27]. We therefore investigated whether the IL-10 release following stimulation of MoDC with scGOS/lcFOS would lead to induction of Tregs. MoDC were treated with scGOS/lcFOS for 24 hours, washed and subsequently co-cultured with naïve T cells in an allogeneic stimulation assay. Compared to control MoDC (medium-treated MoDC control Fig 6A, positive Treg control Fig 6B), scGOS/lcFOS-stimulated DC induced an upward shift in Foxp3 expressing T cells (Fig 6C). However, this increase did not reach significance (Fig 6D).

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