Oral Administration of Zinc Sulfate with Intramuscular Foot-and-Mouth Disease Vaccine Enhances Mucosal and Systemic Immunity

Figures

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  Figure 1. Oral zinc sulfate administration influences early, mid-, and long-term immune responses in mice. (A–E) Overview of experimental strategy (A); antibody titers for SP O measured using PrioCheck™ FMDV kit with undiluted serum (B); antibody titers for SP A assessed using PrioCheck™ FMDV kit with undiluted serum (C); VN titers for O PA2 (D); and VN titers for A YC (E). The dotted line on the y-axis in (B,C) indicates the positive threshold provided by the manufacturer. The dotted line on the y-axis in (D,E) indicates the 1:45 (1.65 Log10) virus-neutralizing titer cutoff level recognized as the defensive level against virus infection. Data are expressed as the mean ± standard error of the mean (SEM) based on triplicate measurements (n = 5/group). Statistical evaluations were conducted using two-way analysis of variance (ANOVA), followed by Tukey’s post hoc test. Significance levels are indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

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  Figure 2. Oral administration of zinc sulfate improves host protection against virus in FMD-vaccinated mice. (A,B) Survival rates post-challenge with O/VET/2013 (A); Changes in body weight post-challenge with O/VET/2013 (B). Data are expressed as mean ± standard error of mean (SEM) based on triplicate measurements (n = 5/group).

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  Figure 3. Oral zinc sulfate administration influences early, mid-term, and long-term immune responses in pigs. (A–E) An overview of the experimental strategy (A); antibody titers for SP O measured using the PrioCheck™ FMDV kit (B); antibody titers for SP A assessed with the PrioCheck™ FMDV kit (C); VN titers for O PA2 (D); and VN titers for A YC (E). The dotted line on the y-axis in (B,C) indicates the positive threshold provided by the manufacturer. The dotted line on the y-axis in (D,E) indicates the 1:45 (1.65 Log10) virus-neutralizing titer cutoff level recognized as the defensive level against virus infection. Data are expressed as the mean ± standard error of the mean (SEM) based on triplicate measurements (n = 5/group). Statistical evaluations were conducted using two-way analysis of variance (ANOVA), followed by Tukey’s post hoc test. Significance levels are indicated as * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001.

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  Figure 4. Oral zinc sulfate administration enhances the expression of secretory IgA. (A,B) sIgA antibody levels in mice (Cusabio® mouse secretory IgA ELISA kit) (A); sIgA antibody levels in pigs (Cusabio® pig secretory IgA ELISA kit) (B). Data are expressed as the mean ± standard error of the mean (SEM) based on triplicate measurements (n = 5/group). Statistical evaluations were conducted using two-way analysis of variance (ANOVA), followed by Tukey’s post hoc test. Significance levels are indicated as * p < 0.05; and **** p < 0.0001.

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  Figure 5. Oral zinc sulfate administration boosts the expression of mucosal immune-related cytokines in pigs. (A–H) Gene expression levels of IL2 (A); IL4 (B); IL12p40 (C); IL17A (D); IL18 (E); IL23p19 (F); IL23R (G); IFNγ (H). Data are expressed as the mean ± standard error of the mean (SEM) based on triplicate measurements (n = 5/group). Statistical evaluations were conducted using two-way analysis of variance (ANOVA), followed by Tukey’s post hoc test. Significance levels are indicated as * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001.