Ig-GAD2 treatment could not overcome overt T1D despite induction of immune tolerance. A: Hyperglycemic (BGL = 160–250 mg/dL) and diabetic (BGL ≥300 mg/dL) NOD mice (six per group) were given Ig-GAD2, and their
BGLs were monitored for a period of 70 days. B and C: The mice of the diabetic group were killed on day 70 and the SP and PLN cells were harvested, stimulated with PMA and ionomycin,
and stained for surface CD4, CD8, and Vβ8.1/8.2 as well as intracellular IFN-γ, IL-10, and IL-17. A group of mice killed upon
diagnosis of diabetes (untreated) was used as control. B: Representative FACS plots (top) and individual mice (bottom) depicting CD4+CD8−Vβ8.1/8.2+ cells producing IFN-γ and IL-10. C: The frequency of CD4+CD8−IL-17+ cells. D: The RQ of mRNA expression for RORγt (rorc) and T-bet (tbx21) in the pancreatic islets of untreated and Ig-GAD2–treated mice. Each bar represents mean ± SEM of three to six mice. *P < 0.05; **P < 0.01.