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. 2017 Sep 4;7:10310. doi: 10.1038/s41598-017-10677-4

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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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Characterisation of IL-10+ lymphocytes. (A) Three-colour flow cytometry showed CD8+ lymphocytes to be the main source of ES-induced IL-10 expression. In contrast, CD4- CD8- double-negative lymphocytes revealed lower IL-10 expression after ES treatment at 150 µg/mL. (B) ES antigens were associated with a mild decrease in Foxp3^high cells within IL-10+ lymphocytes in the absence of LPS stimulation, although the finding failed to reach statistical significance. For AcES at 150 µg/mL, this effect was more obvious when cells were stimulated with LPS (C) Co-staining for IL-17 showed ES antigens to be capable of elevating IL-17 expression in LPS-stimulated IL-10+ lymphocytes and this effect was significant for TcES at 15 µg/mL. Although the data may suggest a decrease in IL-17 expression by unstimulated IL-10+ lymphocytes this effect failed to reach statistical significance. P-values were calculated according to paired t-test. Error bars illustrate standard error of mean (SEM). FSC = forward scatter, SSC = side scatter. Experiments were repeated six times at two different time points.