In Vivo Sustained Release of Peptide Vaccine Mediated by Dendritic Mesoporous Silica Nanocarriers

Figures

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  Figure 1. TEM and DLS analysis of DMSNs-57 and DMSNs-156. TEM image of 57 nm DMSNs (A) and of 156 nm DMSNs (B). Scale bar, 200 nm. The insets show the DMSNs with higher magnification revealing the dendritic structure. Scale bar, 100 nm. (C) DMSNs-57 and DMSNs-156 hydrodynamic size (75 ± 9 nm and 156 ± 10 nm) and (D) ζ-potential values (-30.2 and -37.1 mV).

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  Figure 2. B2T release profiles from B2T@DMSNs-57 and from B2T@DMSNs-156 dispersed in DPBS or BSA-DPBS. After each time point, the supernatants were collected for UV-vis analysis and the pellets were redispersed in the same volume of medium. The procedure was repeated during 1,000 h.

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  Figure 3. RAW 264.7 macrophage cellular interactions of fluoro-B2T@DMSNs-57 and fluoro-B2T@DMSNs-156. (A) CLSM images showing a time- and DMSNs size-dependent internalization. (Note: Green correspond to BodiFluor-488 conjugated to the B2T loaded within the DMSNs whereas the magenta color corresponds to the dye, cell mask deep red used to stain the plasma membrane of the cells). (cf. §SI-11, Figures SI-11, SI-12 , and SI-16 ) (B) Flow cytometry analysis of cellular interactions. The columns represent the mean fluorescence intensity of fluoro-B2T@DMSNs-57 and fluoro-B2T@DMSNs-156. (cf. §SI-2.3, Figures SI – 13 ).

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  Figure 4. In vivo functional validation. ELISA-determined anti-B2T peptide responses of mice vaccinated with B2T@Montanide™ (red circle, n=4), B2T@DMSNs-156nm (green down triangle, n=6), DMSNs-156nm (blue squares, n=4) from sera collected at days 14, 21 (pre-boost) and 40 (post-boost) post-immunization. Each point depicts mean antibody titers (calculated as described in Materials and methods) ± SD for each group. No individual spontaneous reactivity was observed in the titers determined at day 0. (cf. §SI-3; Table SI-1 , and Figures SI-14, SI-15 , and SI-16 ).

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  Figure 5. Sustained in vivo immune response performed by the DMSNs. ELISA-determined anti-B2T peptide responses obtained in vaccination trial II ( Table SI-2 ) of mice vaccinated with B2T@Montanide™ (red circle, n=3), B2T@DMSNs-57nm (purple up triangle, n=5) and B2T@DMSNs-156nm (green down triangle, n=5) from sera collected on the indicated days post-immunization (20, 40, 60, and 80 pi). Each point depicts mean antibody titers (calculated as described in Materials and methods) ± SD for each group. No individual spontaneous reactivity was observed in the titers determined at day 0. (cf. §SI-3; Table SI-2, and Figures SI-14, SI-15, and SI-16 ).