Figure 2.

Immunogenicity of ZnT8 protein and peptides. A and B: ELISPOT assay after immunization and restimulation with the indicated peptides. Shown are the number of IL-2–positive (A) and IFN-γ–positive (B) spot forming cells (SFC) per 10⁶ plated lymph node cells. C and D: ELISPOT after immunization with ZnT8 C-Cyt protein (274–367) and restimulation with the immunizing protein or the peptides indicated. C and D represent number of IL-2–positive and IFN-γ–positive cells, respectively. Blocking of MHC-II was achieved by addition of 5–6 μg/mL anti I-A^g7 antibody. E: IL-2 ELISPOT assay on lymph node cells from NOD mice immunized with the indicated peptide antigen. The cells were restimulated with the immunizing peptide (10 μmol/L) or ZnT8 C-Cyt protein (10 μmol/L), with and without MHC II–blocking antibody (anti I-A^g7). *P values are provided from paired t test. Data are representative of six (A and B), three (C and D), and two (E) experiments. Baseline spots never exceeded 20, were not subtracted, and are indicated in baselines. Bars indicate mean and standard deviation.