Figure 2.

Tregs depletion during boost immunization leads to increased stimulation of IFNγ+Th1 lymphocytes (A), and increased production of IFNγ, IL-2, and IL-6 associated with reduced IL-10 compared to the group without Tregs depletion (B). Vaccination did not stimulate IL-17 production. To analyze the effect of Tregs depletion on the Th1/Th17 response, both male WT littermate and DEREG mice received two subcutaneous injections (0.1 ml) on days 0 and 14 with rSsEno + Gel 01, or PBS alone (non-vaccinated control). Immunized mice were treated with 1 μg of diphtheria toxin (DT) intraperitoneally (i.p.) for two consecutive days, before 24 hours, and during either the first (priming) or the second dose (booster). Immunized and control mice were euthanized 21 days after the primary immunization and their splenocytes were stimulated for 24 hours with 10μg of rSsEno/mL. The supernatant-accumulated cytokines (IL-2, IL-6, IL17A, IFN-γ, and IL-10) were measured by Cytometric Bead Array Th1/Th2/Th17 Cytokine Kit. The results are presented as the mean ± SD of 7 mice per group from two independent experiments, and statistical significance was determined by one-way ANOVA using Tukey’s multiple comparisons tests. *(p < 0.05); **(p < 0.01); ***(p < 0.001); ****(p < 0.0001).