High Throughput Design of Functionalized Nanoparticles for Targeted Vaccine Delivery

This dissertation focuses on the design of functionalized nanoparticles, a novel approach to targeted antigen delivery by decorating the surface of polyanhydride nanoparticles with specific carbohydrates to provide pathogen-like properties. Polyanhydride nanoparticles are promising vaccine platforms that have the potential to be used as controlled drug delivery devices as well as potent and effective immune adjuvants for the design of single dose vaccines. CLRs offer unique advantages for tailoring immune responses. Engagement of CLRs regulates antigen presenting cell activation and promotes delivery of antigens to specific intracellular compartments inside antigen presenting cells for efficient processing and presentation. CLRs recognize conserved carbohydrate structures on the surface of diverse microorganisms, by attaching this carbohydrates to the surface of polyanhydride nanoparticles, the recognition of these nanoparticles and danger signals triggered on antigen presenting cells is similar to the one induced by microorganisms. This strategy could be useful for the rational design of vaccines. In fact, the design of intranasal vaccines is the particular interest of this dissertation, accordingly, the target and engagement of CLRs were studied on alveolar macrophages harvested from mice. The results presented on this work include the development of an efficient high throughput method of synthesis and functionalization of polyanydride nanoparticles; maintaining the morphology of the nanoparticles and a constant concentration of carbohydrates on the surface. In addition, in vitro studies indicate that the co-culture of functionalized nanoparticles with alveolar macrophages significantly increased cell surface expression and pro-inflammatory cytokine production, enhancing alveolar macrophage activation. This phenotype was found to be dependent upon nanoparticle internalization. In conclusion, this study indicate that the strategy of targeting and engaging CLRs is a viable strategy for enhancing the intrinsic adjuvant properties of nanovaccine adjuvants and promoting robust pulmonary immunity