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. 2017 Jul 4;7:4560. doi: 10.1038/s41598-017-04474-2

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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EPA and its metabolite 5-HEPE up-regulate Treg induction in the presence of macrophages. Effects of EPA, fatty acids and EPA metabolites on in vitro induction of Foxp3⁺ regulatory T cells stimulated with CD3ε antibody, adipose tissue macrophages, and TGFβ. Foxp3 expression was assessed by flow cytometry on day six of culture. (A) Effects and dose-dependency of EPA on in vitro Treg induction from Foxp3⁻ T cells isolated from Foxp3 EGFP mice. (B) Effects of the indicated fatty acids (100 μM) on in vitro Treg induction (n = 4). (C) Effects of the indicated purified EPA metabolites (100 μM) on in vitro Treg induction (n = 6). (D) Effects of pharmacological inhibition of the indicated EPA metabolic enzymes on in vitro Treg induction (n = 6). (E) The effect of 20 μM pioglitazone on in vitro Treg induction by splenic dendritic cells and adipose tissue macropahges (n = 3). (F) Isolated CD3⁺ CD4⁺ Foxp3⁻ T cells were co-cultured with adipose tissue macrophages in the presence of GPR119 agonist PSN375963 or GPR120 agonist TUG-891. Percentage of Tregs induced by adipose tissue macrophages in the presence of GPR119 or GPR120 agonist (n = 3). (G) Percentage of Tregs induced by adipose tissue macrophages in the presence of EPA with or without 10 μM GPR119 antagonist TM43718 (n = 3). Data are mean ± SEM. *P < 0.05, **P < 0.01 and ***P < 0.001.