Panel A shows histograms of cell trace violet (CTV) dilution, indicating cell proliferation. The x-axis represents CTV, and the y-axis represents cell count. Different treatments, including PBS, 3A10, and wtIL-2, are compared. Panel B consists of two box plots showing the fold change over PBS in Foxp3 and CTLA-4 mean fluorescence intensity (MFI) in the spleen. The x-axis lists different treatments, and the y-axis shows the fold change. Panel C includes two bar graphs depicting CD25 MFI and Treg number in the pancreatic lymph nodes (pancLN). The x-axis lists treatments, and the y-axis shows the fold change over PBS. Panel D features six bar graphs showing CD25 MFI in host Tregs, CD4, and CD8 cells in the spleen and pancLN. The x-axis lists treatments, and the y-axis shows the fold change over PBS. Panel E is a line graph showing the percentage of diabetes-free mice over time, with different doses of Tregs. The x-axis represents age in weeks, and the y-axis shows the percentage of diabetes-free mice. Different lines represent various doses of Tregs. The graphs collectively illustrate the effects of different treatments on Treg proliferation, marker expression, and diabetes incidence.
OrthoIL-2 3A10 on orthoIL-2R engineered Tregs (related to Fig. 2). (A and B) A mixture of orthoIL-2R–transduced Thy1.1+ and non-transduced Thy1.1− NOD Tregs were transferred to NSG mice followed by daily i.p. injection of PBS, MSA-3A10, or MSA-wtIL-2 (MSA is denoted by *). Spleens were collected on day 3 after Treg transfer for assessing CTV dilution (A) and on day 8 for measuring Foxp3 and CTLA-4 expression (B). Ordinary two-way ANOVA followed by Sidak multiple comparison test was used to determine the statistical significance. (C and D) FACS-purified orthoIL-2(+) BDC2.5 Tregs were transferred to 5-wk-old NOD.CD28KO mice, followed by 7 daily i.p. injection of PBS, MSA-3A10, or MSA-wtIL-2. (C) CD25 expression on orthoIL-2(+) and total number of orthoIL-2R(+) Tregs in the pancLN. (D) CD25 expression on host T cells in the spleen and pancLN. Results shown are summaries of three to five independent experiments. Kruskal–Wallis test was used to determine the statistical significance. (E) A titrated doses of BDC2.5 Tregs were injected into 5-wk-old NOD.CD28KO mice to assess effect on diabetes incidence. Mantel–Cox log-rank test was used to determine the statistical significance. Only statistically significant P values are listed.