J Clin Invest.
Tregs from CT-cured mice can suppress CD4+ T cell activation in an Ag-specific manner.
Cell Proliferation Dye eFluor–labeled (eBioscience) CD4+CD25– Tresps, isolated from normoglycemic NOD mice that had received a prime-boost with human PINS or chicken OVA, were cocultured for 4 days with activated PINS- or OVA-pulsed APCs and the indicated ratios of CD4+CD25+ T cells, isolated from CT-cured mice at the end of the 6-week treatment. (A) Activation of Tresps was measured by flow cytometric analysis of CD44 and shown as MFI normalized to responder-only culture. (B) ELISA measurement of IFN-γ in the supernatant of Treg-Tresp cultures. (C) Percentage of IFN-γ–positive cells in CD4+ Tresps in Treg-Tresp cocultures stimulated with PINS or OVA as indicated. (D) Percentage of IL-10–producing cells in CD4+ Tregs in Treg-Tresp cocultures stimulated with PINS or OVA as indicated. Statistical significance was calculated using Mann-Whitney t test (*P < 0.05, **P < 0.01, ***P < 0.001).