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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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Proliferation of CD4+ T cells with and without blocking of CD83 or PD-1 during DC: T cell co-culture.T cells were co-cultured for 6 days with DCs previously supplemented with fatty acids (50 μM); arachidonic acid (AA), docosahexaenoic acid (DHA), oleic acid (OA) or ethanol only (Ctrl) without (-, No blocking) or with 10 μg/ml purified CD83-antibody (α-CD83) or PD-1 antibody (α-PD-1); and thereafter analyzed by flow cytometry. (A) Proportion of T cells that has proliferated (CellTrace™ CFSElow), with (+) or without (-) PD-1 blocking. (B) Proportion of T cells that has proliferated (CellTrace™ CFSElow), with (+) or without (-) CD83 blocking. 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. Statistical mean difference, for each fatty acid or control, was compared between no blocking and blocking. Data are representative of two experiment. p-values: * <0.05, ** <0.01, *** <0.001, **** <0.0001.
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pone.0143741.g011: Proliferation of CD4+ T cells with and without blocking of CD83 or PD-1 during DC: T cell co-culture.T cells were co-cultured for 6 days with DCs previously supplemented with fatty acids (50 μM); arachidonic acid (AA), docosahexaenoic acid (DHA), oleic acid (OA) or ethanol only (Ctrl) without (-, No blocking) or with 10 μg/ml purified CD83-antibody (α-CD83) or PD-1 antibody (α-PD-1); and thereafter analyzed by flow cytometry. (A) Proportion of T cells that has proliferated (CellTrace™ CFSElow), with (+) or without (-) PD-1 blocking. (B) Proportion of T cells that has proliferated (CellTrace™ CFSElow), with (+) or without (-) CD83 blocking. 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. Statistical mean difference, for each fatty acid or control, was compared between no blocking and blocking. Data are representative of two experiment. p-values: * <0.05, ** <0.01, *** <0.001, **** <0.0001.

Mentions: DCs supplemented with arachidonic acid or DHA had increased expression of PDL-1 and CD83. To evaluate if these markers were involved in the reduced T-cell activation induced by such DCs we added either anti-CD83 or anti-PD-1 blocking antibodies to the DC: T-cell co-cultures. To achieve a more efficient blocking we blocked PD-1 instead of PDL-1, because PD-1 can also bind another molecule, PDL-2. As demonstrated in Fig 11 blocking of PD-1 led to increased proliferation in all DC: T-cell co-cultures. Blocking of PD-1 in arachidonic acid cultures did not restore proliferation to control levels, indicating that other pathways are involved in the low T-cell response. Blocking of CD83 led to lower proliferation in all DC: T-cell co-cultures. This supports the literature describing CD83 as an activation [35] and maturation marker [36].


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)

Proliferation of CD4+ T cells with and without blocking of CD83 or PD-1 during DC: T cell co-culture.T cells were co-cultured for 6 days with DCs previously supplemented with fatty acids (50 μM); arachidonic acid (AA), docosahexaenoic acid (DHA), oleic acid (OA) or ethanol only (Ctrl) without (-, No blocking) or with 10 μg/ml purified CD83-antibody (α-CD83) or PD-1 antibody (α-PD-1); and thereafter analyzed by flow cytometry. (A) Proportion of T cells that has proliferated (CellTrace™ CFSElow), with (+) or without (-) PD-1 blocking. (B) Proportion of T cells that has proliferated (CellTrace™ CFSElow), with (+) or without (-) CD83 blocking. 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. Statistical mean difference, for each fatty acid or control, was compared between no blocking and blocking. Data are representative of two experiment. p-values: * <0.05, ** <0.01, *** <0.001, **** <0.0001.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0143741.g011: Proliferation of CD4+ T cells with and without blocking of CD83 or PD-1 during DC: T cell co-culture.T cells were co-cultured for 6 days with DCs previously supplemented with fatty acids (50 μM); arachidonic acid (AA), docosahexaenoic acid (DHA), oleic acid (OA) or ethanol only (Ctrl) without (-, No blocking) or with 10 μg/ml purified CD83-antibody (α-CD83) or PD-1 antibody (α-PD-1); and thereafter analyzed by flow cytometry. (A) Proportion of T cells that has proliferated (CellTrace™ CFSElow), with (+) or without (-) PD-1 blocking. (B) Proportion of T cells that has proliferated (CellTrace™ CFSElow), with (+) or without (-) CD83 blocking. 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. Statistical mean difference, for each fatty acid or control, was compared between no blocking and blocking. Data are representative of two experiment. p-values: * <0.05, ** <0.01, *** <0.001, **** <0.0001.
Mentions: DCs supplemented with arachidonic acid or DHA had increased expression of PDL-1 and CD83. To evaluate if these markers were involved in the reduced T-cell activation induced by such DCs we added either anti-CD83 or anti-PD-1 blocking antibodies to the DC: T-cell co-cultures. To achieve a more efficient blocking we blocked PD-1 instead of PDL-1, because PD-1 can also bind another molecule, PDL-2. As demonstrated in Fig 11 blocking of PD-1 led to increased proliferation in all DC: T-cell co-cultures. Blocking of PD-1 in arachidonic acid cultures did not restore proliferation to control levels, indicating that other pathways are involved in the low T-cell response. Blocking of CD83 led to lower proliferation in all DC: T-cell co-cultures. This supports the literature describing CD83 as an activation [35] and maturation marker [36].

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