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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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Correlation between proliferation and expression of regulatory markers on T cells.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. Proportion of divided cells was correlated to proportion of T cells double-positive for FoxP3 and (A) Helios, (B) CTLA-4, and (C) PD-1. All samples, regardless of stimuli, were used in the same correlation analysis. The proportion of divided cells is different in (C) compared to (A) and (B) since this staining was performed in another experiment. r value: non-parametric Spearman correlation.
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pone.0143741.g009: Correlation between proliferation and expression of regulatory markers on T cells.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. Proportion of divided cells was correlated to proportion of T cells double-positive for FoxP3 and (A) Helios, (B) CTLA-4, and (C) PD-1. All samples, regardless of stimuli, were used in the same correlation analysis. The proportion of divided cells is different in (C) compared to (A) and (B) since this staining was performed in another experiment. r value: non-parametric Spearman correlation.

Mentions: In addition we divided the DO11.10+ T cells into FoxP3+ and FoxP3neg cells (S6A Fig) and as stated above, there was a significantly higher proportion of FoxP3+cells with both arachidonic acid and DHA relative to control (S6B Fig). The FoxP3+ T cells did not differ in MFI for FoxP3 depending on fatty acid-priming of the co-cultured DC (S6C Fig), meaning that arachidonic acid and DHA resulted in a higher proportion of FoxP3+ cells but not a higher expression of FoxP3 molecules on each cell. We further determined the expression of Helios, CTLA-4 and PD-1 by proportion (%) and MFI on both the FoxP3+ (S6D Fig) and FoxP3neg T-cell population (S6E Fig). Phenotypically the FoxP3+ population were similar in all cultures i.e. the MFI values and proportions of Helios, CTLA-4 and PD-1 were similar between the different fatty acids and control (S6D and S6E Fig). As noted above, CTLA-4 was expressed to a lower extent in the FoxP3neg population in both arachidonic and DHA compared to control cultures (S6E Fig). There was a negative correlation between proliferation (% divided cells) and proportion of Tregs assessed by the following populations; FoxP3+ Helios+ (Spearman r-value -0.92, p<0.0001), FoxP3+ CTLA-4+ (r = - 0.94, p<0.0001) and FoxP3+ PD-1+ (r = -0.63, p = 0.0034) (Fig 9). In summary, DCs supplemented with arachidonic acid and, to some extent, DHA inhibit T-cell proliferation, accompanied by a higher proportion of Tregs. These Tregs have a similar phenotype as Tregs found in oleic acid or control cultures.


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)

Correlation between proliferation and expression of regulatory markers on T cells.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. Proportion of divided cells was correlated to proportion of T cells double-positive for FoxP3 and (A) Helios, (B) CTLA-4, and (C) PD-1. All samples, regardless of stimuli, were used in the same correlation analysis. The proportion of divided cells is different in (C) compared to (A) and (B) since this staining was performed in another experiment. r value: non-parametric Spearman correlation.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0143741.g009: Correlation between proliferation and expression of regulatory markers on T cells.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. Proportion of divided cells was correlated to proportion of T cells double-positive for FoxP3 and (A) Helios, (B) CTLA-4, and (C) PD-1. All samples, regardless of stimuli, were used in the same correlation analysis. The proportion of divided cells is different in (C) compared to (A) and (B) since this staining was performed in another experiment. r value: non-parametric Spearman correlation.
Mentions: In addition we divided the DO11.10+ T cells into FoxP3+ and FoxP3neg cells (S6A Fig) and as stated above, there was a significantly higher proportion of FoxP3+cells with both arachidonic acid and DHA relative to control (S6B Fig). The FoxP3+ T cells did not differ in MFI for FoxP3 depending on fatty acid-priming of the co-cultured DC (S6C Fig), meaning that arachidonic acid and DHA resulted in a higher proportion of FoxP3+ cells but not a higher expression of FoxP3 molecules on each cell. We further determined the expression of Helios, CTLA-4 and PD-1 by proportion (%) and MFI on both the FoxP3+ (S6D Fig) and FoxP3neg T-cell population (S6E Fig). Phenotypically the FoxP3+ population were similar in all cultures i.e. the MFI values and proportions of Helios, CTLA-4 and PD-1 were similar between the different fatty acids and control (S6D and S6E Fig). As noted above, CTLA-4 was expressed to a lower extent in the FoxP3neg population in both arachidonic and DHA compared to control cultures (S6E Fig). There was a negative correlation between proliferation (% divided cells) and proportion of Tregs assessed by the following populations; FoxP3+ Helios+ (Spearman r-value -0.92, p<0.0001), FoxP3+ CTLA-4+ (r = - 0.94, p<0.0001) and FoxP3+ PD-1+ (r = -0.63, p = 0.0034) (Fig 9). In summary, DCs supplemented with arachidonic acid and, to some extent, DHA inhibit T-cell proliferation, accompanied by a higher proportion of Tregs. These Tregs have a similar phenotype as Tregs found in oleic acid or control cultures.

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