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. 2020 May 20;10:8350. doi: 10.1038/s41598-020-65212-9

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© The Author(s) 2020

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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Infection-experienced and naïve T_regs are equally well maintained in vivo. CD4+GFP+ T_regs were sorted from naive (CD45.1) or LCMV-experienced (CD45.2) Foxp3-GFP reporter mice and 500’000 of each population were adoptively co-transferred i.v. into naive CD90.1+ recipient mice. Organs from T_reg recipients were harvested and analyzed by flow cytometry 21–28 days after transfer. (A) Characterization of T_regs from naive and LCMV-experienced mice before transfer. Input ratio (top) and purity (bottom) are shown in representative plots. (B) On day 21–28 post transfer, expression of CD25, CXCR3, CD44, TIGIT and Ki67 was determined in naive or LCMV-experienced CD4+Foxp3+ T_regs isolated from the spleens of recipient mice. Representative plots (top) and summary plots (bottom) are depicted. (C) Chimerism (left) and absolute numbers (right) of CD90.2⁺ T_regs recovered from the indicated organs on day 21 to 28 post transfer. (B,C) Cumulative data (Mean ± SD; biological replicates: Spleen n = 7, pLNs n = 4, BM n = 4, Lung n = 3; 4 independent experiments) (t Test, *p < 0.05, **p < 0.01, ***p < 0.001).