Figure 2.

The inability of NSG recipients to respond to common gamma chain cytokines is associated with an ability of NOD donor polyclonal T-cells to impair T1D transfer by AI4 CD8 T-cells.

A) Total splenocytes from NOD-scid or NSG mice (2.5×10⁵) were cultured in vitro in the presence of 10 pg/ml IL-2 and with or without 0.1% NaN₃ acting as inhibitor of cytokine translocation. After three days in culture, the amount of IL-2 remaining in the supernatant was quantified by ELISA. B) NSG and NOD-scid mice were injected i.v. with a mixture of 2.5×10⁶ NOD-AI4 and 5×10⁶ standard NOD splenocytes. Data indicate T1D incidence from 2 independent experiments total n=20 for each strain. Disease incidence compared by Log-rank (Mantel-Cox) Test (*** p<0.0001). C, D) At 2 weeks post-transfer the total number and activation phenotype of AI4 CD8⁺ T-cells in each type of recipient (n=3 and 4 respectively for NOD-scid and NSG recipients) was analyzed by flow cytometry in spleen (SPL), pancreatic lymph node (PLN), and mesenteric lymph node (MLN). In A, C, and D bars represent mean ± SEM, * p<0.05, *** p<0.0001, ns p>0.05, Two-way ANOVA with Bonferroni post-test correction.