<p>SMNCs secreting IFN-γ, IL-4, or IL-10 upon stimulation were detected by ELISPOT assay. Groups of six mice were immunized intramuscularly three times at 2-week intervals using 10 µg of NM2e protein in normal saline (NS) or 10 µg of NM2e formulated with Al(OH)<sub>3</sub>, CpG 1826, or Al(OH)<sub>3</sub> plus CpG 1826. Mice immunized with NS or Al(OH)<sub>3</sub> plus CpG 1826 were treated as negative controls. All mice in each treatment group were sacrificed on day 38. SMNCs were separated from mouse spleen samples, and 5 µg/ml NP<sub>147–155</sub>, NP<sub>55–69</sub>, and M2e peptide pool were used as stimulants in the ELISPOT assays. The numbers of SMNCs producing IFN-γ (A), IL-4 (B), or IL-10 (C) after stimulation for 40 h with NP<sub>147–155</sub> (left), NP<sub>55–69</sub> (middle), or M2e peptides (right) are presented as spot-forming cells (SFCs)/10<sup>6</sup> SMNCs. Columns show the average SFCs/10<sup>6</sup> SMNCs, and bars indicate the standard deviation of each group. Lines above two or more groups indicate that they have the same comparative results. *, <i>p</i>≤0.05; **, <i>p</i>≤0.01; ***, <i>p</i>≤0.001 by one-way ANOVA.</p
