Immunostimulant-integrated acrylate nanocomposite adjuvants potentiate foot-and-mouth disease vaccine efficacy

Figures

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  Fig. 1. Physicochemical properties of the AHN and AHN@X adjuvants: (a) Appearance; (b) Mean particle diameter and P.I.; (c-d) Storage stability at ambient temperatures of 4 °C and 37 °C; (e) Particle size distribution and zeta potential; (f) Micromorphology.

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  Fig. 2. Safety of adjuvant: (a) Cytotoxicity of macrophages treated with AHN and AHN@X for 24 h, as assessed by CCK8 assay (n = 5, data were analyzed using two-way ANOVA); (b) Weight monitoring of FMDV, AHN and AHN@X with time of initial immunization (n = 6). Data are presented as the mean ± SD. ⁎P < 0.05, ⁎⁎P < 0.01.

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  Fig. 3. Specific antibody response of FMDV, AHN and AHN@X measured by ELISA on days 14 and 28 post-immunization: (a) IgG antibody titer; (b) IgG1 antibody titer; (c) IgG2a antibody titer; (d) IgG2a/IgG1 ratio. Data were analyzed using two-way ANOVA and presented as mean ± SD. N = 6, ⁎P < 0.05, ⁎⁎P < 0.01.

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  Fig. 4. Cytokine secretion levels of spleen cells stimulated by FMDV (5 μg/mL) for 24 h on day 28 post-immunization: (a) IL-4; (b) IFN-γ; (c) IFN-γ/IL-4 ratio. Data were analyzed using one-way ANOVA and presented as mean ± SD. n = 3, ⁎P < 0.05, ⁎⁎P < 0.01.

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  Fig. 5. Percentages of B cells and T cells in spleen cells stimulated by FMDV (5 μg/mL) for 24 h were analyzed by flow cytometry on day 28 post-immunization: Activation and representative flow cytometry of lymphocytes in each group of mice on day 28 post-immunization: (a) B cell activation; (b) CD4+ T cell activation; (c) Central memory CD4+ T cells; (d) Effector memory CD4+ T cells; (e) Representative flow cytometry. Data were analyzed using one-way ANOVA and presented as mean ± SD. n = 3, ⁎P < 0.05, ⁎⁎P < 0.01.

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  Fig. 6. Endocytosis capacity of macrophages treated with FITC-OVA, AHN/FITC-OVA, and AHN@X/FITC-OVA for 24 h, as measured by an inverted fluorescence microscope. Green: FITC-OVA, blue: nuclei stained with DAPI. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)