Figure 4.

Cytokine releases from splenocytes of immunized mice. IFN-γ, IL-12, and IL-4 secretion in splenocytes was evaluated following stimulation with CFP-10:Fcγ2a, CFP-10:His, and PHA. Mice were immunized with CFP-10:Fcγ2a in DDA/TDM, and CFP-10:His in DDA/TDM, respectively. PBS and BCG were used as controls. Freshly isolated spleen cells were plated in duplicate at 5×10⁶ cell per well in 24-well plate and incubated with CFP-10:Fcγ2a and CFP-10:His (10 μg/ml) and PHA (5μg/mL) for 72 h at 37 °C, 5 % CO₂. IFN-γ, IL-12, and IL-4 were detected using mouse IFN-γ, IL-12, and IL-4 ELISA kits. A and B) CFP-10:Fcγ2a stimulated splenic cells from immunized mice with CFP-10:Fcγ2a had the highest numbers of IFN-γ and IL-12 secretion than CFP-10:His, BCG, or PBS immunized groups (P <0.05). C) Both CFP-10:Fcγ2a- and CFP-10:His immunized groups showed increased level of IL-4 production, as a marker of Th2, compare to BCG and PBS control groups (P<0.05). D) Differential IFN-γ, IL-12 and IL-4 production in the vaccinated mice showed all immunizes groups were pushed toward Th1 responses. *There is no significant difference between the groups with at least one shared letter