Splenic T-cell responses. Sorted CD4⁺ T cells were stimulated with PMA and ionomycin for 8 hours. Brefeldin A was added for the last 4 hours. The cells were then fixed and permeabilized for IFN-γ, IL-17, and IL-4 intracellular staining. (A) Th1 cells were defined as CD4⁺IFN-γ⁺, Th17 cells as CD4⁺IL-17⁺, and Th2 cells as CD4⁺IL-4⁺. Dot plots are of one representative RTX-treated ITP patient. (B) Data are summarized in dot plots representing the different T-cell subsets (Th1, Th17, and Th2) in controls (n = 7), RTX-untreated ITP patients (ITP RTX⁻; n = 7), and RTX-treated patients (ITP RTX⁺; n = 7). The balance between immune response and immune tolerance was evaluated by assessing the Th1/Treg and Th17/Treg ratios. The Th1/Th2 ratio is also represented. The horizontal bars represent the mean values with standard deviations. *P < .05; **P < .001; ***P < .0001; NS, nonsignificant.