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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

Suppressive capacity of T cells induced by scGOS/lcFOS-treated MoDC.T cells of allogeneic stimulation assays (ASA) were harvested on day seven and co-incubated with freshly isolated, violet-labeled responder CD4+ T cells a ratio of 1:1 for five days in presence of anti-CD3/28 and rhIL-2. The mix of ASA cells and violet stained responder CD4+ T cells was then stained (after five days of co-incubation) with CD4 APC-Cy7. The dot plots show data from one representative experiment. Proliferation of responder T cells only (A) or in co-incubation with T cells induced by medium- (B), LPS- (C) and scGOS/lcFOS- (E) stimulated MoDC and Treg control (D) is shown. Results in (F) are presented as mean ± SEM, 4 independent experiments are depicted. Proliferation of violet-positive cells was analyzed by flow cytometry and suppressive functionality index was determined by calculating dividing cells (div)/ non-dividing (ndiv) cells (F). * p<0.05, Mann Whitney test. ASA = allogeneic stimulation assay, scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharides, MoDC = monocyte-derived dendritic cells, div = dividing cells, ndiv = non-dividing cells.
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pone.0132304.g007: Suppressive capacity of T cells induced by scGOS/lcFOS-treated MoDC.T cells of allogeneic stimulation assays (ASA) were harvested on day seven and co-incubated with freshly isolated, violet-labeled responder CD4+ T cells a ratio of 1:1 for five days in presence of anti-CD3/28 and rhIL-2. The mix of ASA cells and violet stained responder CD4+ T cells was then stained (after five days of co-incubation) with CD4 APC-Cy7. The dot plots show data from one representative experiment. Proliferation of responder T cells only (A) or in co-incubation with T cells induced by medium- (B), LPS- (C) and scGOS/lcFOS- (E) stimulated MoDC and Treg control (D) is shown. Results in (F) are presented as mean ± SEM, 4 independent experiments are depicted. Proliferation of violet-positive cells was analyzed by flow cytometry and suppressive functionality index was determined by calculating dividing cells (div)/ non-dividing (ndiv) cells (F). * p<0.05, Mann Whitney test. ASA = allogeneic stimulation assay, scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharides, MoDC = monocyte-derived dendritic cells, div = dividing cells, ndiv = non-dividing cells.

Mentions: These induced CD4+Foxp3+ T cells differentiated by scGOS/lcFOS-stimulated MoDC showed in suppression assays an increased capacity to inhibit the proliferation of CD4+ responder cells (Fig 7A and 7E) compared to T cells induced by medium-/LPS-treated MoDC and Treg control (Fig 7B–7D). As demonstrated in Fig 7F these scGOS/lcFOS-induced Foxp3+ T cells are characterized by a low suppressive functionality index comparable to Treg control which indicates a high capacity to inhibit the proliferation of CD4+ responder cells.


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)

Suppressive capacity of T cells induced by scGOS/lcFOS-treated MoDC.T cells of allogeneic stimulation assays (ASA) were harvested on day seven and co-incubated with freshly isolated, violet-labeled responder CD4+ T cells a ratio of 1:1 for five days in presence of anti-CD3/28 and rhIL-2. The mix of ASA cells and violet stained responder CD4+ T cells was then stained (after five days of co-incubation) with CD4 APC-Cy7. The dot plots show data from one representative experiment. Proliferation of responder T cells only (A) or in co-incubation with T cells induced by medium- (B), LPS- (C) and scGOS/lcFOS- (E) stimulated MoDC and Treg control (D) is shown. Results in (F) are presented as mean ± SEM, 4 independent experiments are depicted. Proliferation of violet-positive cells was analyzed by flow cytometry and suppressive functionality index was determined by calculating dividing cells (div)/ non-dividing (ndiv) cells (F). * p<0.05, Mann Whitney test. ASA = allogeneic stimulation assay, scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharides, MoDC = monocyte-derived dendritic cells, div = dividing cells, ndiv = non-dividing cells.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0132304.g007: Suppressive capacity of T cells induced by scGOS/lcFOS-treated MoDC.T cells of allogeneic stimulation assays (ASA) were harvested on day seven and co-incubated with freshly isolated, violet-labeled responder CD4+ T cells a ratio of 1:1 for five days in presence of anti-CD3/28 and rhIL-2. The mix of ASA cells and violet stained responder CD4+ T cells was then stained (after five days of co-incubation) with CD4 APC-Cy7. The dot plots show data from one representative experiment. Proliferation of responder T cells only (A) or in co-incubation with T cells induced by medium- (B), LPS- (C) and scGOS/lcFOS- (E) stimulated MoDC and Treg control (D) is shown. Results in (F) are presented as mean ± SEM, 4 independent experiments are depicted. Proliferation of violet-positive cells was analyzed by flow cytometry and suppressive functionality index was determined by calculating dividing cells (div)/ non-dividing (ndiv) cells (F). * p<0.05, Mann Whitney test. ASA = allogeneic stimulation assay, scGOS/lcFOS = short chain galacto-, long chain fructo-oligosaccharides, MoDC = monocyte-derived dendritic cells, div = dividing cells, ndiv = non-dividing cells.
Mentions: These induced CD4+Foxp3+ T cells differentiated by scGOS/lcFOS-stimulated MoDC showed in suppression assays an increased capacity to inhibit the proliferation of CD4+ responder cells (Fig 7A and 7E) compared to T cells induced by medium-/LPS-treated MoDC and Treg control (Fig 7B–7D). As demonstrated in Fig 7F these scGOS/lcFOS-induced Foxp3+ T cells are characterized by a low suppressive functionality index comparable to Treg control which indicates a high capacity to inhibit the proliferation of CD4+ responder cells.

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