Journal: Nature Communications
Article Title: Efficient oral vaccination by bioengineering virus-like particles with protozoan surface proteins
Figure Lengend Snippet: Induction of anti-HA humoral and cellular responses in mice vaccinated with VLP-HA/VSP-G. a – e Mice were orally vaccinated with VLPs and the humoral response against HA in different biological samples was evaluated. Levels of total serum anti-HA IgG ( a ), IgG1 and IgG2a ( b ), n = 8 from two independent experiments. Levels of IgA anti-HA in fecal extracts and BAL, n = 6 from two independent experiments ( c ). Levels of serum IgG anti-HA measured at different times post immunization, n = 6 from two independent experiments ( d ). Levels of serum IgG anti-HA in animals immunized with a mixture of VLP-HA plus VLP-VSP-G, n = 6 from two independent experiments ( e ). f Levels of serum IgG and fecal IgA in mice vaccinated with VLPs via oral or subcutaneous (s.c.) route, n = 8 from two independent experiments. g Mice were orally immunized with four weekly doses of 50 μg of recombinant ΔVSP1267 or vehicle. One week after the last immunization the presence of anti-VSP1267 antibodies in blood and fecal samples was checked, then these animals were orally immunized with VLP-HA/VSP-G, according to the protocol previously used. One week after the last VLP dose, the levels of serum IgG and fecal IgA anti-HA Abs were measured, n = 6 from two independent experiments. h , i Mice were orally vaccinated with VLPs and the cellular response against HA was evaluated. The frequency of HA-specific IFN-γ-secreting T cells (spot-forming colonies, SFC) was determined after antigen-specific re-stimulation of splenocytes, n = 7 from two independent experiments ( h ). Cytokines were measured in splenocyte supernatants and only those cytokines with detectable levels are shown, n = 6 from two independent experiments ( i ). Data were analyzed by one-way ANOVA and Tukey’s multiple comparison test ( a – f , h , i ) or by two-way ANOVA and Bonferroni post-tests ( g ). Values represent mean ± s.e.m. * p
Article Snippet: TEM of cryosections was performed by applying the Tokuyasu technique and incubated with anti-HA or anti-VSP and anti-mouse coupled 5-nm gold particles (Sigma-Aldrich, Cat. # G7527, dilution 1/50).
Techniques: Mouse Assay, Recombinant