Figure 5.

Vaccination with αDC-SIGN:P25 plus αCD40, CTA1-DD or zymosan induces pro-inflammatory cytokine production. (A) Experimental scheme. WT or hSIGN mice were vaccinated i.p. at days −42, −28, and −14 with vehicle (PBS) or αDC-SIGN:P25 (10 µg) plus αCD40 (10 µg), CTA1-DD (10 µg) or zymosan (200 µg). At day 0, mice were challenged i.v. with 2 × 10⁶ Mycobacterium bovis BCG-Ag85B. Five days later, mice were sacrificed and splenocytes were isolated and stained for intracellular cytokine production after ex vivo restimulation with P25 (30 µg/mL). (B) Representative flow cytometry plots depicting the percentage of IFN-γ⁺ (upper panels), IL-2⁺ (middle panels) and TNF-α⁺ (lower panels) among CD44⁺ CD4⁺ T cells. (C–E) Graphs represent percent of IFN-γ⁺, IL-2⁺, TNF-α⁺ and polyfunctional (IFN-γ⁺ IL-2⁺ TNF-α⁺) among CD44⁺ CD4⁺ T cells of mice vaccinated with (C) αCD40, (D) CTA1-DD and (E) zymosan. Each symbol represents an individual mouse and results are pooled from two experiments with 4–5 mice per group. *p < 0.05, **p < 0.01, and ***p < 0.001; two-way ANOVA with Bonferroni’s post hoc test.