Figure 4.

Mixed vaccination induces weak lung Th1 and splenic Th1 and Tc1 responses. Mice were immunized SC two times with 100 µL of NCMX, and the third booster was administered via intranasal instillation. Twenty-one days after the last immunization, the spleen and lungs were collected and analyzed by flow cytometry for Th1, Th17, and Tc1 lymphocytes (A). The number of Th1 (CD4⁺IFN-γ⁺), Th17 (CD4⁺IL-17⁺), and Tc1 (CD8⁺IFN-γ⁺) cells in the spleen (B, D and F, respectively) and lungs (C, E and G, respectively) are shown. Serum samples were collected, and the humoral immune response was evaluated by measuring the levels of IgG1 (H) and IgG2a (I). Differences between the means of the groups were determined by Student's t-test, and p values are shown. Significant differences were found between the groups, n = 4.