Figure 3.

The role of Salmonella-based combination therapy induced Tregs in suppression of autoimmune diabetes. Female 7–8 week old NOD mice were treated with 2.5 μg/mouse of anti-CD3 mAb via i.p. injection for 5 days followed by 2 doses of oral combined vaccine. Splenocyte pools were prepared from the indicated treated groups at day 30 post-vaccination. (A) Adoptive transfer of splenocytes from overtly diabetic NOD mice into recipient NSG mice. (B,E) Adoptive transfer of splenocyte pools from NOD mice treated with vehicle or combined therapy into NSG mice. (C,F) Co-transfer of splenocyte Tregs from vehicle or vaccinated mice with diabetic splenocytes into NSG recipient mice. (D,G) Adoptive transfer of Treg-depleted splenocytes isolated from vehicle or vaccinated NOD mice into NSG mice. (A–D) Data display the blood glucose levels for individual mice over the time. (E–G) Data presented as the Log-rank plot of the percentage of NSG mice that remained diabetes-free over the time after transfer. The differences between the group of mice vaccinated with combined therapy and vehicle was significant (*p < 0.05, ****p < 0.0001) by the log-rank (Mantel-Cox) test.