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The Polyunsaturated Fatty Acids Arachidonic Acid and Docosahexaenoic Acid Induce Mouse Dendritic Cells Maturation but Reduce T-Cell Responses In Vitro.

Carlsson JA, Wold AE, Sandberg AS, Östman SM - PLoS ONE (2015)

Bottom Line: Here, we measured the effects of omega-3 (n-3), n-6 and n-9 fatty acids on the interaction between dendritic cells (DCs) and naïve T cells.Fatty acids were taken up by the DCs, as shown by gas chromatography analysis.However, fewer T cells co-cultured with these DCs proliferated (CellTrace Violet low) and expressed CD69 or CD25, while more were necrotic (7AAD+).

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases, Institute of Biomedicine, the Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.

ABSTRACT
Long-chain polyunsaturated fatty acids (PUFAs) might regulate T-cell activation and lineage commitment. Here, we measured the effects of omega-3 (n-3), n-6 and n-9 fatty acids on the interaction between dendritic cells (DCs) and naïve T cells. Spleen DCs from BALB/c mice were cultured in vitro with ovalbumin (OVA) with 50 μM fatty acids; α-linolenic acid, arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid or oleic acid and thereafter OVA-specific DO11.10 T cells were added to the cultures. Fatty acids were taken up by the DCs, as shown by gas chromatography analysis. After culture with arachidonic acid or DHA CD11c+ CD11b+ and CD11c+ CD11bneg DCs expressed more CD40, CD80, CD83, CD86 and PDL-1, while IAd remained unchanged. However, fewer T cells co-cultured with these DCs proliferated (CellTrace Violet low) and expressed CD69 or CD25, while more were necrotic (7AAD+). We noted an increased proportion of T cells with a regulatory T cell (Treg) phenotype, i.e., when gating on CD4+ FoxP3+ CTLA-4+, CD4+ FoxP3+ Helios+ or CD4+ FoxP3+ PD-1+, in co-cultures with arachidonic acid- or DHA-primed DCs relative to control cultures. The proportion of putative Tregs was inversely correlated to T-cell proliferation, indicating a suppressive function of these cells. With arachidonic acid DCs produced higher levels of prostaglandin E2 while T cells produced lower amounts of IL-10 and IFNγ. In conclusion arachidonic acid and DHA induced up-regulation of activation markers on DCs. However arachidonic acid- and DHA-primed DCs reduced T-cell proliferation and increased the proportion of T cells expressing FoxP3, indicating that these fatty acids can promote induction of regulatory T cells.

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Co-expression of FoxP3 with Helios, CTLA-4 and PD-1.T cells were co-cultured for 6 days with dendritic cells (DCs) previously supplemented with fatty acids (50 μM); arachidonic acid (AA), docosahexaenoic acid (DHA), oleic acid (OA) or ethanol only (Ctrl); and thereafter analyzed by flow cytometry. (A) Representative dot plots from control samples showing staining of FoxP3+ and indicated markers (left column). In each plot double-positive-cells are found in the upper right quadrant and double-negative cells in the lower left quadrant. FMO samples are shown in the right column. (B) Proportion of double-positive (right column), FoxP3+ samples (middle column) and FoxP3- samples (left column) in CD4+ cells. Each dot represents one individual. Black dots denote samples supplemented with fatty acid while white dots with black borders denote control (ethanol only). Horizontal black solid lines show median value. The median from the control group has been extended with a dotted line for easy comparison to the other groups. Statistical mean difference was compared to the control group. Data are representative of two independent experiments. p-values: * <0.05, ** <0.01, *** <0.001, **** <0.0001.
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pone.0143741.g008: Co-expression of FoxP3 with Helios, CTLA-4 and PD-1.T cells were co-cultured for 6 days with dendritic cells (DCs) previously supplemented with fatty acids (50 μM); arachidonic acid (AA), docosahexaenoic acid (DHA), oleic acid (OA) or ethanol only (Ctrl); and thereafter analyzed by flow cytometry. (A) Representative dot plots from control samples showing staining of FoxP3+ and indicated markers (left column). In each plot double-positive-cells are found in the upper right quadrant and double-negative cells in the lower left quadrant. FMO samples are shown in the right column. (B) Proportion of double-positive (right column), FoxP3+ samples (middle column) and FoxP3- samples (left column) in CD4+ cells. Each dot represents one individual. Black dots denote samples supplemented with fatty acid while white dots with black borders denote control (ethanol only). Horizontal black solid lines show median value. The median from the control group has been extended with a dotted line for easy comparison to the other groups. Statistical mean difference was compared to the control group. Data are representative of two independent experiments. p-values: * <0.05, ** <0.01, *** <0.001, **** <0.0001.

Mentions: Since arachidonic acid and DHA were the only fatty acids that affected DC phenotype as well as the DCs’ ability to induce T-cell proliferation and activation markers, we chose to focus on these and included oleic acid as an inert fatty acid control. To investigate if the low T-cell proliferation observed with arachidonic acid- and DHA-primed DCs could be linked to presence of Tregs in the cultures we stained the T cells with FoxP3, Helios, CTLA-4 and PD-1. For representative dot plots and gating strategy see Fig 8A. We found that the proportion of FoxP3+ DO11.10+ T cells, either co-expressing Helios, CTLA-4 or PD-1 or not, were higher in DC: T cell co-cultures in which the DCs had been primed with arachidonic acid, and to some extent also DHA, compared to control cultures (Fig 8B). In contrast the proportion of FoxP3neg DO11.10+ T cells co-expressing CTLA-4 or PD-1 was lower (Fig 8B). With oleic acid-primed DCs the proportion of putative Tregs was similar as in the control cultures. If proliferation mainly occurred among effector T cells, and not Tregs, the higher proportion of Tregs with arachidonic acid and DHA might be due to limited proliferation in these cultures and not because of regulatory activity. Expression of Helios, CTLA-4 and PD-1 on the whole T cell population is shown in S5 Fig.


The Polyunsaturated Fatty Acids Arachidonic Acid and Docosahexaenoic Acid Induce Mouse Dendritic Cells Maturation but Reduce T-Cell Responses In Vitro.

Carlsson JA, Wold AE, Sandberg AS, Östman SM - PLoS ONE (2015)

Co-expression of FoxP3 with Helios, CTLA-4 and PD-1.T cells were co-cultured for 6 days with dendritic cells (DCs) previously supplemented with fatty acids (50 μM); arachidonic acid (AA), docosahexaenoic acid (DHA), oleic acid (OA) or ethanol only (Ctrl); and thereafter analyzed by flow cytometry. (A) Representative dot plots from control samples showing staining of FoxP3+ and indicated markers (left column). In each plot double-positive-cells are found in the upper right quadrant and double-negative cells in the lower left quadrant. FMO samples are shown in the right column. (B) Proportion of double-positive (right column), FoxP3+ samples (middle column) and FoxP3- samples (left column) in CD4+ cells. Each dot represents one individual. Black dots denote samples supplemented with fatty acid while white dots with black borders denote control (ethanol only). Horizontal black solid lines show median value. The median from the control group has been extended with a dotted line for easy comparison to the other groups. Statistical mean difference was compared to the control group. Data are representative of two independent experiments. p-values: * <0.05, ** <0.01, *** <0.001, **** <0.0001.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4664484&req=5

pone.0143741.g008: Co-expression of FoxP3 with Helios, CTLA-4 and PD-1.T cells were co-cultured for 6 days with dendritic cells (DCs) previously supplemented with fatty acids (50 μM); arachidonic acid (AA), docosahexaenoic acid (DHA), oleic acid (OA) or ethanol only (Ctrl); and thereafter analyzed by flow cytometry. (A) Representative dot plots from control samples showing staining of FoxP3+ and indicated markers (left column). In each plot double-positive-cells are found in the upper right quadrant and double-negative cells in the lower left quadrant. FMO samples are shown in the right column. (B) Proportion of double-positive (right column), FoxP3+ samples (middle column) and FoxP3- samples (left column) in CD4+ cells. Each dot represents one individual. Black dots denote samples supplemented with fatty acid while white dots with black borders denote control (ethanol only). Horizontal black solid lines show median value. The median from the control group has been extended with a dotted line for easy comparison to the other groups. Statistical mean difference was compared to the control group. Data are representative of two independent experiments. p-values: * <0.05, ** <0.01, *** <0.001, **** <0.0001.
Mentions: Since arachidonic acid and DHA were the only fatty acids that affected DC phenotype as well as the DCs’ ability to induce T-cell proliferation and activation markers, we chose to focus on these and included oleic acid as an inert fatty acid control. To investigate if the low T-cell proliferation observed with arachidonic acid- and DHA-primed DCs could be linked to presence of Tregs in the cultures we stained the T cells with FoxP3, Helios, CTLA-4 and PD-1. For representative dot plots and gating strategy see Fig 8A. We found that the proportion of FoxP3+ DO11.10+ T cells, either co-expressing Helios, CTLA-4 or PD-1 or not, were higher in DC: T cell co-cultures in which the DCs had been primed with arachidonic acid, and to some extent also DHA, compared to control cultures (Fig 8B). In contrast the proportion of FoxP3neg DO11.10+ T cells co-expressing CTLA-4 or PD-1 was lower (Fig 8B). With oleic acid-primed DCs the proportion of putative Tregs was similar as in the control cultures. If proliferation mainly occurred among effector T cells, and not Tregs, the higher proportion of Tregs with arachidonic acid and DHA might be due to limited proliferation in these cultures and not because of regulatory activity. Expression of Helios, CTLA-4 and PD-1 on the whole T cell population is shown in S5 Fig.

Bottom Line: Here, we measured the effects of omega-3 (n-3), n-6 and n-9 fatty acids on the interaction between dendritic cells (DCs) and naïve T cells.Fatty acids were taken up by the DCs, as shown by gas chromatography analysis.However, fewer T cells co-cultured with these DCs proliferated (CellTrace Violet low) and expressed CD69 or CD25, while more were necrotic (7AAD+).

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases, Institute of Biomedicine, the Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.

ABSTRACT
Long-chain polyunsaturated fatty acids (PUFAs) might regulate T-cell activation and lineage commitment. Here, we measured the effects of omega-3 (n-3), n-6 and n-9 fatty acids on the interaction between dendritic cells (DCs) and naïve T cells. Spleen DCs from BALB/c mice were cultured in vitro with ovalbumin (OVA) with 50 μM fatty acids; α-linolenic acid, arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid or oleic acid and thereafter OVA-specific DO11.10 T cells were added to the cultures. Fatty acids were taken up by the DCs, as shown by gas chromatography analysis. After culture with arachidonic acid or DHA CD11c+ CD11b+ and CD11c+ CD11bneg DCs expressed more CD40, CD80, CD83, CD86 and PDL-1, while IAd remained unchanged. However, fewer T cells co-cultured with these DCs proliferated (CellTrace Violet low) and expressed CD69 or CD25, while more were necrotic (7AAD+). We noted an increased proportion of T cells with a regulatory T cell (Treg) phenotype, i.e., when gating on CD4+ FoxP3+ CTLA-4+, CD4+ FoxP3+ Helios+ or CD4+ FoxP3+ PD-1+, in co-cultures with arachidonic acid- or DHA-primed DCs relative to control cultures. The proportion of putative Tregs was inversely correlated to T-cell proliferation, indicating a suppressive function of these cells. With arachidonic acid DCs produced higher levels of prostaglandin E2 while T cells produced lower amounts of IL-10 and IFNγ. In conclusion arachidonic acid and DHA induced up-regulation of activation markers on DCs. However arachidonic acid- and DHA-primed DCs reduced T-cell proliferation and increased the proportion of T cells expressing FoxP3, indicating that these fatty acids can promote induction of regulatory T cells.

Show MeSH
Related in: MedlinePlus