(A-B) Splenic CD11c+ DCs were pulsed with 1 μg/ml CBirTox or CBir1 peptide for 1 hour while CD19+ B cells or YAMC cells were pulsed with 2 μg/ml CBirTox or CBir peptide for 2 or 4 hours, respectively, and co-cultured with CD4+CD25- CBir1 Tg T cells for 4 days before flow cytometry analysis. Flow plots (A) are representative of 3–6 independent experiments; aggregate data shown (B). (C-D) CD19+ B cells were treated similarly to A and co-cultured with CD4+CD25- CBir1 Tg T cells for 4 days before stimulation with PMA and ionomycin in the presence of Golgi Stop for 4 hours prior to staining for intracellular cytokines. Flow plots (C) are representative of 4 independent experiments. (D) Results are expressed as the mean ± SEM. *p<0.05, **p<0.005, ***p<0.0005, NS, not significant. Groups of three were analyzed using one-way ANOVA with Bonferroni’s post test, while groups of two were analyzed using Student’s unpaired t test.