Fig. 3. Promotion of antigen-specific immune responses by PPTEE-glucan in C57BL/6 mice. C57BL/6 mice were vaccinated with OVA-containing microemulsion (ME) every 2 weeks for a total of 3 vaccinations. Serum was collected 21 days post-vaccine injection, and the titer and Area Under Curve (AUC) of OVA-specific IgM (A), IgG1 (B), IgG2b (C), and IgG2c (D) in the serum were analyzed by ELISA. (E) The AUC ratio of OVA-specific IgG1 to OVA-specific IgG2b was calculated (E). At 35 d post-vaccine injection, the spleens of vaccinated mice were extracted, and splenocytes were stimulated with an OVA-containing growth medium for 3 days. IFN-γ-expressing CD4+T cells and CD8+T cells were analyzed using flow cytometry (F). For the in vivo CD8+T cell proliferation assay, C57BL/6 recipient mice were injected with QS-21- and PPTEE-containing ME the day after transferring CD45.1 OT-I CD8+T cells. Three days after ME injection, CD45.1 OT-I CD8+T cells in the spleen were analyzed by flow cytometry (G). The data is presented as mean ± SEM. Statistical analysis was conducted using the unpaired t-test for (E) and one-way ANOVA for (A-D, F, G), with significance: non-significant (ns) when p≥0.05, *p<0.05, **p<0.01, ***p<0.001.
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