Figure 5.

The induction of CD8⁺ Tregs by Cal/BDP and the suppression of the Foxp3 expression in CD4⁺ Tregs by BDP. Mice received the topical application of IMQ cream on the ear for 6 consecutive days and were treated with 0.6 nmol Cal, 0.06 nmol BDP, or the combination of 0.6 nmol Cal and 0.06 nmol BDP 1 h before each IMQ application. (A–C) Changes in the proportion of CD4⁺ Tregs (gated on CD45⁺CD3⁺CD4⁺CD25⁺Foxp3⁺), CD8⁺ Tregs (gated on CD45⁺CD3⁺CD8⁺CD122⁺PD-1⁺), and Bregs (gated on CD45⁺B220⁺CD5⁺CD1d⁺IL-10⁺) in the CD45⁺ leukocyte population. The rates of CD4⁺ Tregs, CD8⁺ Tregs, and Bregs in the CD45⁺ leukocyte population were analyzed by flow cytometry 1 day after the final IMQ application. (D–F) The expression of Foxp3 in the CD4⁺ Treg subset, PD-1 in the CD8⁺ Treg subset, and IL-10 in the Breg subset in dLNs. The geometric means of Foxp3 in the CD4⁺ Treg subset, of PD-1 in the CD8⁺ Treg subset, and of IL-10 in the Breg subset in dLNs were analyzed by flow cytometry 1 day after the final IMQ application. Each symbol represents an individual mouse, and the horizontal lines indicate the means of the groups (n = 6). *p < 0.05, **p < 0.01, ***p < 0.001 by Tukey test among the vehicle, Cal, BDP, and Cal/BDP groups.