Fig. 5

GLS-1027 prevents type 1 diabetes in the NOD mouse. (A) Eight to nine weeks-old female NOD mice were treated with GLS-1027 (5 mg/mouse) under a late prophylactic regime, from 10 to 20 weeks of age (n = 16 per group). Blood glucose was monitored and mice with blood glucose levels ≥ 11 mmol/L on two consecutive occasions were considered diabetic. Diabetic mice were sacrificed the day when diagnosis was confirmed. (B) Spleens and pancreatic lymph nodes were aseptically isolated from the mice and cells from spleen (10⁶/well) or lymph nodes (5 × 10⁵/well) were cultured with Concanavalin A (Con A; 2.5 μg/ml) in the of culture medium for 48 h. The concentration of IL-17A in the culture media was measured by ELISA. (C) Serum samples were obtained from terminal blood samples collected immediately after CO₂ euthanasia and assessed for IL-17A levels via ELISA. (D) Spleens and pancreatic lymph nodes were aseptically isolated from the mice and cells from spleen (10⁶/well) or lymph nodes (5 × 10.⁵/well) were cultured with Concanavalin A (Con A; 2.5 μg/ml) in 1 ml of culture medium for 48 h. The concentration of TNF-α in the culture media was measured by ELISA. (E) Serum samples were obtained from terminal blood samples collected immediately after CO₂ euthanasia and assessed for TNF-α levels via ELISA. Treatment with GLS-1027 resulted in a dose-dependent inhibition of cytokines. Error bars in all panels indicate 1 standard deviation among 3 replicates per condition