A Subunit Vaccine Based on E2 Protein of Atypical Porcine Pestivirus Induces Th2-type Immune Response in Mice

Figures

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  Figure 1. Western blot analysis of the expression of atypical porcine pestivirus (APPV) E2 protein. (A) Western blot analysis of APPV E2 protein expressed in Sf9 cells infected with the recombinant baculovirus AC-3-APPV-E2. Lane 1: Culture supernatant of sf9 cells infected with Ac-3-APPV-E2; Lane 2: Lysates of Sf9 cells infected with Ac-3-APPV-E2. (B) APPV E2 protein digested with PNGase F by Western blot analysis. Lane 1: APPV E2 protein as normal control; Lane 2: APPV E2 protein digested with PNGase F.

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  Figure 2. SDS-PAGE analysis of the purified recombinant APPV E2 protein. Lanes 1–8, purified recombinant APPV E2 protein eluted with five column volumes of 80 mM imidazole.

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  Figure 3. APPV E2-specific IgG detected by indirect ELISA at different dpi. Data represent the mean ± standard error of mean (SEM). Different letters (a, b, c) indicate a statistically significant difference between different experimental groups (p < 0.05).

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  Figure 4. IgG subtype in serum samples tested using indirect ELISA. (A) APPV E2-specific IgG1 level. (B) APPV E2-specific IgG2a level. (C) APPV E2-specific IgG3 level. Data represent the mean ± SEM. Different letters (a, b) indicate a statistically significant difference between different experimental groups (p < 0.05).

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  Figure 5. Detection of lymphocyte proliferative responses. Data represent the mean ± SEM. Different letters (a, b) indicate a statistically significant difference between different experimental groups (p < 0.05).

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  Figure 6. Detection of cytokine production in the supernatants of stimulated lymphocytes from mice immunized with the APPV E2 subunit vaccines. Concentrations of IFN-γ (A), IL-2 (B), IL-4 (C), and IL-10 (D) detected with commercial ELISA kits in accordance with the manufacturer’s instructions. Data represent the mean ± SEM. Different letters (a, b) indicate a statistically significant difference between different experimental groups (p < 0.05).