Fig. 3

mESC-TEP-treated recipients have an increased number and function of Tregs. NOD mice were injected i.v. with anti-CD3/CD8 Abs on days − 8 and − 3. On day 0, the conditioned mice were injected i.v. with BM and CD4⁺ T cell-depleted spleen cells from MHC-matched H-2^g7 B6 mice. Groups of the BMT recipient were also injected i.t. with MHC-mismatched B6 mESC- EpCAM1⁺ TEPs or mESC-control cells (EpCAM1⁻ cells) as in Fig. 2. On day 100, Tregs in the thymus and spleen were analyzed by flow cytometry. a Representative flow cytometric profiles showing thymic Tregs. b The number of thymic CD4⁺CD8⁻CD3⁺FoxP3⁺ Tregs from each group. c Representative flow cytometric profiles showing the Tregs in the spleen of the recipients. d The number of splenic CD4⁺CD25⁺FoxP3⁺ Tregs from each group. e CD4⁺CD25⁺ Tregs were purified from the spleen of each group. CD4⁺CD25⁻ Teffs were purified from wild-type NOD mice. The Teffs were stimulated with anti-CD3 antibody (0.5 μg/ml) in the presence or absence of Tregs at a ratio 2:1 for 3 days. The proliferation of Teffs was measured by [³H] thymidine incorporation. The results are presented as suppression index. c–e The data are expressed as mean ± SD from one of three independent experiments with similar results (4–5 mice per group in each experiment). *P < 0.05, compared with mice given MHC-matched BMT only