Determination of CD4+ T cells activation ex vivo after immunization. The determination of three different activation markers, CD69, CD71, and CD25, was performed ex vivo in mice immunized with each of the recombinant BCG vaccine candidates. For rBCG-N-SARS-CoV-2, the activation of CD69 (A), CD71 (B), and CD25 (C) were evaluated in CD4+ T cells. For rBCG-N-ANDV, the activation of CD69 (D), CD71 (E), and CD25 (F) were evaluated in CD4+ T cells. Data are representative of 3 independent experiments. Each experiment considered a total of three mice per group. Statistical differences were assessed by two-way ANOVA with a post hoc Tukey test. *: p < 0.05; **: p < 0.01; ***: p < 0.001.