pTreg generation and maintenance by antigen feeding. (A) Number of DO CD4⁺ T cells in dLNs in Fig. 1 A. (B) Frequency of IFN-γ⁺, IL-4⁺, and IL-17A⁺ cells among DO CD4⁺ T cells in dLNs (n = 10 and 11). (C) CD25 and Foxp3 expression by DO CD4⁺ T cells in the spleen at 4 wk after designated feeding (n = 4). (D) Changes in left ear thickness in Fig. 1 A. Arrowheads indicate OVA challenges. (E) Histology and histological scores of the left ear in NC- and EWP-fed DORe mice. Bars in histology indicate 100 µm. (F and G) Frequencies of pTreg cells in mLNs, dLNs, and the right ear (F) and frequencies of IFN-γ⁺, IL-4⁺, and IL-17A⁺ cells in dLNs (G) in the mice shown in Fig. 1 G (n = 7 [PBS-treated] or n = 6 [DTx-treated]). (H and I) Frequencies of pTreg cells in mLNs and dLNs (H), and frequencies of IFN-γ⁺, IL-4⁺, and IL-17A⁺ cells in dLNs (I) in the mice shown in Fig. 1 J (n = 4–6). (J and K) Frequencies of pTreg cells in mLNs and dLNs (J), and frequencies of IFN-γ⁺, IL-4⁺, and IL-17A⁺ cells in dLNs (K) (n = 4–6). Vertical bars indicate the mean ± SEM. Statistical significance was assessed by an unpaired t test or Welch’s t test (A–C, E–G, J, and K), and the Tukey–Kramer test (H and I). P values were shown in the figures.