FIG. 3.
CA treatment reduces insulin-specific T-cell effector function and frequency. NOD mice were treated with CA or HBSS daily. Mice were immunized with insulin in CFA. A: Inguinal LNs were removed at day 6 postimmunization and surface stained for CD4 as well as intracellularly stained for IFN-γ for flow cytometric analysis. Cells were gated on CD4+ cells (n = 3 independent experiments with two mice per group). **P < 0.005. B: Inguinal LNs were isolated on day 6 and stimulated with insulin in a recall IFN-γ ELISPOT. Two days after stimulation, ELISPOT plates were developed, and spots were counted using the Zeiss KS Elispot Imaging system. Frequency = 2.5 × 105/average number of spots per treatment. Graph shows the average of three independent experiments performed in triplicate. *P < 0.05. MFI, mean fluorescence intensity.