Figure 5: B7.1wa-DC and multiligand-DC treatments at early-hyperglycemic stage results in reversal of hyperglycemia for significant durations.

Early-hyperglycemic female NOD mice (blood glucose: 140–250 mg/dl) were treated i.v. with control-DCs and B7.1wa-DCs and multiligand-DCs twice as described for Fig. 4. pCDH1-multiligand vector, instead of individual ligand vectors, was used for generating multiligand-DCs in this experiment. A) Blood glucose levels were examined every week and glucose values of individual mice are shown. Dotted line represents overt-hyperglycemic (blood glucose: >450 mg/dl). This experiment was carried out in 2 batches of 4 mice/group and the cumulative results are shown. B) Cohorts of mice were euthanized 30-days post treatment, and pancreatic sections were subjected to insulin (green), glucagon (red) and DAPI (blue) staining and insulin positive islets and immune cell infiltration, based on DAPI staining, were scored as detailed under materials and method section. *Pancreatic tissues from untreated mice that were at early-hyperglycemic stage at the time of euthanasia were included as “none” control group to assess the pre-treatment frequency of insulin positive islets. Pancreatic tissues from at least 5 mice were used for each group. C) PnLN cells from individual mice (6 mice/group), cultured for 24h with BcAg, and injected (i.v.) into NOD-Rag1−/− mice (about 1×10⁶ cells/mouse; one recipient/donor) and examined for blood glucose levels every week, for up to 20 weeks. P-value by log-rank test with overt hyperglycemia for two consecutive weeks as the end-point for panel A, Fisher’s exact test for comparing grades ≤3+ and ≥4+ among groups for panel B, and log-rank test for panel C. All P-values are in comparison with control-DC group.