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. 2018 Jan 11;8:420. doi: 10.1038/s41598-017-17981-z

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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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The different Treg subsets exhibit unique capacities to undergo in vitro expansion with maintained FoxP3 expression and suppressive activity. (A) The number of T cells that were produced after a 14d in vitro stimulation is shown (n = 3). The expanded T cell populations were tested for (B) expression of intracellular FoxP3 to indicate purity; and (C) for their ability to suppress CFSE labeled mTresp when co-cultured at a 1:1/2 (mTresp:Treg) ratio with αCD3/APC stimulation. (D) Shows representative intracellular staining of the expanded Tregs for co-expression of FoxP3 and IL-17. (E) The frequency of IL-17 producing, FoxP3+ cells in the DR⁻mTreg versus the nvTreg populations (n = 4) is shown. Data shown are mean ± SEM. Significance, determined by Student’s t test, is indicated by (*P < 0.05), (**P < 0.01), and (***P < 0.001).