Characterization of nanoparticles for vaccine development
Nanotechnology are becoming to play an important role in vaccine development, it offers the opportunity to design nanoparticles varying in composition, size, shape and surface properties for application in the fields of medicine. In this work, we aimed to characterize nanoparticles as a safety adjuvant for further use in immunotherapies. Polycationic polymer nanoparticles (R3N0) were used in BMDC to characterize them in vitro. Nanoparticle-cell interaction (internalization or adhesion) was evaluated by microscopy and cell phenotype activation (MHCII and CD86) were determined by flow-cytometry. IL-1β-production was also evaluated by ELISA. Furthermore, Balb/c mice were intraperitoneally sensitized with R3N0, R3N0 plus OVA (combined) and OVA conjugated to R3N0. Finally, humoral (serum antibodies) and cellular immune response (cytokines) was evaluated by ELISA and flow cytometry. We found that R3N0 is internalized by the cells generating its activation with an increase in the expression of CD86 and IL-1β. In turn, R3N0 promotes grater stimulation and release of IL-1β than the positive control (P<0.05). IL-1β was abrogated by the presence of inhibitors of the pathways that activate the inflammosome NLRP3. In vivo, we observed that the OVA-specific-IgG levels were much higher than those corresponding to the adjuvant used as a positive control (P<0.05). Immunized mice showed an increase of IFN-γ by LTCD4 and LTCD8. In conclusion, we found that R3N0 are internalized by BMDC and activates them with the production of proinflammatory cytokines. Moreover, this nanoparticle may have a potential adjuvant effect with the production of Th1 specific antibodies and an IFN-γ dependent cell response.