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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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Activation of T cells by fatty-acid supplemented dendritic cells (DCs).T cells were co-cultured for 6 days with DCs previously supplemented with fatty acids (50 μM); α-linolenic acid (ALA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA), oleic acid (OA) or ethanol only (Ctrl); and thereafter analyzed by flow cytometry. (A) Gating strategy for lymphocytes (upper row), DO11.10+ T cells (middle row) and divided DO11.10+ T cells; CellTrace™ Violetlow (bottom row). (B) Representative dot plots showing dividing and activated (CD25+ and CD69+) DO11.10+ cells for arachidonic acid (AA), DHA and control samples. (C) Proportion divided, (D) CD25+ and (E) CD69+ of DO11.10+ T 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 solid black 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. p-values: ** <0.01, *** <0.001, **** <0.0001.
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pone.0143741.g006: Activation of T cells by fatty-acid supplemented dendritic cells (DCs).T cells were co-cultured for 6 days with DCs previously supplemented with fatty acids (50 μM); α-linolenic acid (ALA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA), oleic acid (OA) or ethanol only (Ctrl); and thereafter analyzed by flow cytometry. (A) Gating strategy for lymphocytes (upper row), DO11.10+ T cells (middle row) and divided DO11.10+ T cells; CellTrace™ Violetlow (bottom row). (B) Representative dot plots showing dividing and activated (CD25+ and CD69+) DO11.10+ cells for arachidonic acid (AA), DHA and control samples. (C) Proportion divided, (D) CD25+ and (E) CD69+ of DO11.10+ T 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 solid black 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. p-values: ** <0.01, *** <0.001, **** <0.0001.

Mentions: Next, we investigated if the fatty acid exposure of DCs would alter the outcome of the antigen-presentation to naïve T cells. Therefore, DCs were cultured with fatty acid and the model antigen OVA for 3 days, thereafter OVA-specific DO11.10+ T cells were added to the DC cultures (S1 Fig). The gating strategy of T cells is shown in Fig 6A and 6B. Briefly, lymphocytes were gated (upper 3 panels, Fig 6A), followed by gating for DO11.10 expression (middle panel, Fig 6A). Proliferation is shown in the lower panel of Fig 6A. After culture with OVA, about 25% of the T cells had divided, only 2% had divided in the absence of antigen. The solvent, ethanol had an inhibitory effect on proliferation, i.e. about 20% of the T cells proliferated in control cultures (Fig 6A). Arachidonic acid- or DHA-primed DCs reduced subsequent T-cell proliferation, compared to the control DCs, from ≈ 20% to ≈ 5% (Fig 6C). Expression of the activation markers CD69 and CD25 was reduced in parallel (Fig 6D and 6E). There was a correlation between proliferation and expression of CD69 (r = 0.8587, p < 0.0001) as well as proliferation and CD25 (r = 0.8935, p < 0.0001) (S4 Fig). None of the other investigated fatty acids (linoleic acid, α-linolenic acid, oleic acid or EPA) suppressed the capacity to activate T cells.


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)

Activation of T cells by fatty-acid supplemented dendritic cells (DCs).T cells were co-cultured for 6 days with DCs previously supplemented with fatty acids (50 μM); α-linolenic acid (ALA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA), oleic acid (OA) or ethanol only (Ctrl); and thereafter analyzed by flow cytometry. (A) Gating strategy for lymphocytes (upper row), DO11.10+ T cells (middle row) and divided DO11.10+ T cells; CellTrace™ Violetlow (bottom row). (B) Representative dot plots showing dividing and activated (CD25+ and CD69+) DO11.10+ cells for arachidonic acid (AA), DHA and control samples. (C) Proportion divided, (D) CD25+ and (E) CD69+ of DO11.10+ T 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 solid black 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. p-values: ** <0.01, *** <0.001, **** <0.0001.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4664484&req=5

pone.0143741.g006: Activation of T cells by fatty-acid supplemented dendritic cells (DCs).T cells were co-cultured for 6 days with DCs previously supplemented with fatty acids (50 μM); α-linolenic acid (ALA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA), oleic acid (OA) or ethanol only (Ctrl); and thereafter analyzed by flow cytometry. (A) Gating strategy for lymphocytes (upper row), DO11.10+ T cells (middle row) and divided DO11.10+ T cells; CellTrace™ Violetlow (bottom row). (B) Representative dot plots showing dividing and activated (CD25+ and CD69+) DO11.10+ cells for arachidonic acid (AA), DHA and control samples. (C) Proportion divided, (D) CD25+ and (E) CD69+ of DO11.10+ T 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 solid black 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. p-values: ** <0.01, *** <0.001, **** <0.0001.
Mentions: Next, we investigated if the fatty acid exposure of DCs would alter the outcome of the antigen-presentation to naïve T cells. Therefore, DCs were cultured with fatty acid and the model antigen OVA for 3 days, thereafter OVA-specific DO11.10+ T cells were added to the DC cultures (S1 Fig). The gating strategy of T cells is shown in Fig 6A and 6B. Briefly, lymphocytes were gated (upper 3 panels, Fig 6A), followed by gating for DO11.10 expression (middle panel, Fig 6A). Proliferation is shown in the lower panel of Fig 6A. After culture with OVA, about 25% of the T cells had divided, only 2% had divided in the absence of antigen. The solvent, ethanol had an inhibitory effect on proliferation, i.e. about 20% of the T cells proliferated in control cultures (Fig 6A). Arachidonic acid- or DHA-primed DCs reduced subsequent T-cell proliferation, compared to the control DCs, from ≈ 20% to ≈ 5% (Fig 6C). Expression of the activation markers CD69 and CD25 was reduced in parallel (Fig 6D and 6E). There was a correlation between proliferation and expression of CD69 (r = 0.8587, p < 0.0001) as well as proliferation and CD25 (r = 0.8935, p < 0.0001) (S4 Fig). None of the other investigated fatty acids (linoleic acid, α-linolenic acid, oleic acid or EPA) suppressed the capacity to activate T cells.

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