FIG. 3.

Rapamycin/IL-2 combination therapy results in a persistent enhancement of IL-2 responsiveness in CD4⁺CD25⁺ T cells. Thawed PBMCs were stimulated with media alone or 100 IU/mL IL-2 prior to fixation, permeabilization, and staining for CD4, CD25, and pSTAT5. Representative dot plots of CD4⁺ cells are shown in A. B: The frequency of pSTAT5⁺ cells of CD4⁺CD25⁺ cells (A, top) is shown for control (n = 5) and T1D (n = 5) subjects matched to subjects in the trial (noted by ITN R + 2 Trial) by HLA, age, sex, and genotype (PTPN221858 or PTPN2rs1893217). Day 0 and 28 analyses are shown for patients in the trial. Statistical significance was determined between control and T1D subjects (matched + trial) using a two-sample Student t test and between day 0 and 28 in the trial using a paired Student t test. C: The frequency and fold increase (FI) of pSTAT5 in CD4⁺CD25⁺ cells are shown over time for patients in the trial. Statistical significance was determined by a one-way ANOVA and is shown in both graphs. Significance from time 0 to each time point was determined using a paired Student t test and is indicated with an asterisk. *P ≤ 0.05, **P ≤ 0.01, and ***P ≤ 0.001.