Foxp3+ Regulatory T Cells in Mouse Models of Type 1 Diabetes
Adoptive TCR-HA+ T cell transfer into immunocompetent RIP-HA mice. (a) Using the clonotypic antibody 6.5, naïve TCR-HA+ T cells ( CD25−GFP−) were FACS purified from BALB/c.Thy1.1 mice, after CD4 bead enrichment of pooled cells from spleen and LNs. Presort (top) and postsort (bottom) analyses of TCR-HA/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, TCR-HA+ T cells were cultured as previously described , in the presence of peptide (10 g/mL). As indicated, naïve or in vitro preactivated TCR-HA+ T cells were injected i.v. into immunocompetent BALB/c.Thy1.2 RIP-HA mice. (b) Flow cytometry of expression among gated CD4+Thy1.1+ cells at day 14 after adoptive transfer into BALB/c.RIP-HA recipient mice (mLN: mesenteric lymph node; pLN: pancreatic LN). Numbers in dot plots or histograms indicate the percentage of cells in the respective gate. (c) Blood glucose concentrations (top) and diabetes incidence (bottom) of BALB/c.RIP-HA mice injected with naïve (blue circles, ) or in vitro preactivated TCR-HA+ T cells (red circles, ). Blood glucose concentrations (in mg/dL) of individual mice were determined every other day and plotted against time. The grey dashed line indicates normoglycemia. Mice were considered diabetic at blood glucose levels above 200 mg/dL (red dashed line) on at least two consecutive measurements or with blood glucose levels once above 400 mg/dL.