Figure 2.

Ag85B-specific CD4⁺ T cell response. C57BL/6 mice were subcutaneously immunized as summarized in Figure 1. Draining lymph nodes were collected 7 and 28 days after priming, and 3 and 10 days after booster immunization and analyzed for the frequency of Ag-specific CD4⁺ T cells, identified by staining with Ag85B-specific MHC class II tetramers (Tet-Ag85B). (A) Scatter plot of CD44 versus Tet-Ag85B, gated on live CD4⁺ lymphocytes, shown from a single animal representative of each immunization group, collected 10 days after boosting. Activated tetramer⁺ cells are gated and the frequency reported. (B) Time course analysis of the frequencies of tetramer⁺ CD4⁺ T cells, detected in each group, reported as mean ± SEM of five mice per group. Mann–Whitney test for multiple pairwise comparisons was used for assessing statistical difference between groups receiving the same booster and primed with H56 alone or H56 + CAF01 adjuvant. Kruskal–Wallis test, followed by Dunn’s post test for multiple comparison, was used to assess statistical difference among all groups (P ≤ 0.05).