Figure 2. Selective dysregulation of Th2 responses in Itch^fl/flFoxp3^Cre mice.

(A) The percentage and number of CD4⁺ and CD8⁺ T cells in the spleen (sp) of Itch^fl/flFoxp3^Cre (white squares) and Itch^+/+Foxp3^Cre (black circles) littermates. (B) Flow cytometric analysis of CD44, CD62L, and CD69 expression on conventional CD4⁺ T cells in 8-week-old Itch^fl/flFoxp3^Cre mice and Itch^+/+Foxp3^Cre littermates. One representative experiment of three independent experiments is shown. (C) Flow cytometric analysis of cytokine production by splenic CD4⁺ T cells from 8-week-old Itch^fl/flFoxp3^Cre mice and Itch^+/+Foxp3^Cre littermates. Splenocytes were stimulated with PMA (100 ng/ml) and ionomycin (1 μg/ml) in the presence of Golgi stop for 4 hours. (D) Frequency of CD4⁺ T cells producing IL-4, IL-5, IFN-γ, IL-17, IL-2, or IL-10. Data represent the mean ± SEM. (E) Serum samples were obtained from 8- to 12-week-old Itch^fl/flFoxp3^Cre mice and Itch^+/+Foxp3^Cre littermates (n = 6). Itch^fl/flFoxp3^Cre (white squares) and Itch^+/+Foxp3^Cre mice (black circles). Total amounts of each Ig subclass were determined by ELISA using isotype-specific antibodies. *P < 0.05; **P < 0.01; ***P < 0.001 (unpaired 2-tailed Student’s t test).