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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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Distribution of dendritic cells (DCs) in fatty-acid supplemented cell cultures.DC cultures were supplemented with fatty acids (50 μM); α-linolenic acid (ALA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA) or oleic acid (OA) for 3 days and thereafter DCs were analyzed by flow cytometry. (A) Dot plots showing gating strategy for CD11c+ CD11bneg (lower gate) and CD11c+ CD11b+ (upper gate) DCs based on FMO (fluorescence minus one) samples. To the right a representative sample, in the middle FMO for CD11b and to the right FMO for CD11c. (B) Proportion of CD11c+ DCs, both CD11bneg and CD11b+, in fatty-acid supplemented cell cultures. (C) Proportion of CD11b+ cells within the CD11c+ population shown in Fig 2B. Black bars denote samples supplemented with fatty acid while white bar with black border denotes control (ethanol only). For each group n = 8. Error bars show standard deviation.
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pone.0143741.g002: Distribution of dendritic cells (DCs) in fatty-acid supplemented cell cultures.DC cultures were supplemented with fatty acids (50 μM); α-linolenic acid (ALA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA) or oleic acid (OA) for 3 days and thereafter DCs were analyzed by flow cytometry. (A) Dot plots showing gating strategy for CD11c+ CD11bneg (lower gate) and CD11c+ CD11b+ (upper gate) DCs based on FMO (fluorescence minus one) samples. To the right a representative sample, in the middle FMO for CD11b and to the right FMO for CD11c. (B) Proportion of CD11c+ DCs, both CD11bneg and CD11b+, in fatty-acid supplemented cell cultures. (C) Proportion of CD11b+ cells within the CD11c+ population shown in Fig 2B. Black bars denote samples supplemented with fatty acid while white bar with black border denotes control (ethanol only). For each group n = 8. Error bars show standard deviation.

Mentions: After 3 days of in vitro culture with fatty acids DCs were analyzed by flow cytometry for separation of DC subsets and examination of their costimulatory molecule expression (Experimental design, S1 Fig). The FACS analysis revealed a high background in both the CD11c and CD11b channel (Fig 2A), reflecting low purity and pronounced autofluorescence of in vitro cultured cells. CD11c+ cells were gated, based on higher fluorescence than to the FMO control (Fig 2A) and divided into CD11c+ CD11b+ (upper gate) and CD11c+ CD11bneg (lower gate) subpopulations, shown in Fig 2A. This was made because the relative level of expression for several cell surface molecules were higher in the CD11b+ group, as seen in the histograms in Figs 3 and 4. The proportion of CD11c+ cells (Fig 2B) or CD11c+ CD11b+ cells (Fig 2C) did not differ between cultures supplemented with fatty acids, compared to control cultures with ethanol only.


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)

Distribution of dendritic cells (DCs) in fatty-acid supplemented cell cultures.DC cultures were supplemented with fatty acids (50 μM); α-linolenic acid (ALA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA) or oleic acid (OA) for 3 days and thereafter DCs were analyzed by flow cytometry. (A) Dot plots showing gating strategy for CD11c+ CD11bneg (lower gate) and CD11c+ CD11b+ (upper gate) DCs based on FMO (fluorescence minus one) samples. To the right a representative sample, in the middle FMO for CD11b and to the right FMO for CD11c. (B) Proportion of CD11c+ DCs, both CD11bneg and CD11b+, in fatty-acid supplemented cell cultures. (C) Proportion of CD11b+ cells within the CD11c+ population shown in Fig 2B. Black bars denote samples supplemented with fatty acid while white bar with black border denotes control (ethanol only). For each group n = 8. Error bars show standard deviation.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0143741.g002: Distribution of dendritic cells (DCs) in fatty-acid supplemented cell cultures.DC cultures were supplemented with fatty acids (50 μM); α-linolenic acid (ALA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA) or oleic acid (OA) for 3 days and thereafter DCs were analyzed by flow cytometry. (A) Dot plots showing gating strategy for CD11c+ CD11bneg (lower gate) and CD11c+ CD11b+ (upper gate) DCs based on FMO (fluorescence minus one) samples. To the right a representative sample, in the middle FMO for CD11b and to the right FMO for CD11c. (B) Proportion of CD11c+ DCs, both CD11bneg and CD11b+, in fatty-acid supplemented cell cultures. (C) Proportion of CD11b+ cells within the CD11c+ population shown in Fig 2B. Black bars denote samples supplemented with fatty acid while white bar with black border denotes control (ethanol only). For each group n = 8. Error bars show standard deviation.
Mentions: After 3 days of in vitro culture with fatty acids DCs were analyzed by flow cytometry for separation of DC subsets and examination of their costimulatory molecule expression (Experimental design, S1 Fig). The FACS analysis revealed a high background in both the CD11c and CD11b channel (Fig 2A), reflecting low purity and pronounced autofluorescence of in vitro cultured cells. CD11c+ cells were gated, based on higher fluorescence than to the FMO control (Fig 2A) and divided into CD11c+ CD11b+ (upper gate) and CD11c+ CD11bneg (lower gate) subpopulations, shown in Fig 2A. This was made because the relative level of expression for several cell surface molecules were higher in the CD11b+ group, as seen in the histograms in Figs 3 and 4. The proportion of CD11c+ cells (Fig 2B) or CD11c+ CD11b+ cells (Fig 2C) did not differ between cultures supplemented with fatty acids, compared to control cultures with ethanol only.

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