Journal of Diabetes Research / 2013 / Article / Fig 2

Review Article

Foxp3+ Regulatory T Cells in Mouse Models of Type 1 Diabetes

Figure 2

Adoptive BDC2.5+ T cell transfer into immunodeficient NOD mice. (a) Using anti-V 4 antibodies, naïve BDC2.5+ T cells CD25GFP) were FACS purified from NOD.Foxp ×BDC2.5 mice, after CD4 bead enrichment of pooled cells from spleen and LNs. Presort (top) and postsort (bottom) analyses of V 4/CD62L (left) and CD25/GFP (right) expression among CD4-gated cells are depicted. The gating scheme is illustrated by the line with arrowhead. For antigen-specific stimulation in vitro, BDC2.5+ T cells were cultured as previously described [7], in the presence of the mimotope peptide RTRPLWVRME (10  g/mL). Naïve or in vitro preactivated BDC2.5+ T cells were injected i.v. into NOD.Rag1−/− recipient mice, as indicated below. (b) Flow cytometry of GFP (left) and CD25/GFP (right) expression among gated CD4+ cells from LNs at day 15 after adoptive transfer into NOD.Rag1−/− recipient mice. Numbers in dot plots indicate the percentage of cells in the respective quadrant or gate. (c) Blood glucose concentrations (top) and diabetes incidence (bottom) of NOD.Rag1−/− recipient mice injected with naïve (5 × 105 cells/mouse, blue triangles, ) or in vitro preactivated BDC2.5+ T cells (5 × 105 cells/mouse, red squares, ; or 6×106 cells/mouse, red circles, ). Blood glucose concentrations of recipient mice were determined and plotted as described in the legend for Figure 1.