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The aim of this thesis was to formulate nanoparticles with DNA plasmid (pDNA), encoding the Mycobacterium tuberculosis (Mtb) antigen 85A. The design of cationic N-trimethylated chitosan (TMC) nanoparticles, adjuvanted with the pattern recognition ligands TLR-9 and NLR-2, is a novel approach for improving DNA vaccines. To provide a potential context for future design of DNA vaccines, important parameters of TMC nanoparticle formulations with pDNA were evaluated and characterized. Successful binding of pDNA to the nanoparticles was confirmed as well as integrity of the secondary structure of pDNA after being released. It was shown that two pattern-recognition ligands co-delivered by nanoparticles synergistically activate innate immune responses in vitro and enhance cell-mediated immune responses in vivo. TMC nanoparticles, formulated with Mtb antigen 85A expressing pDNA, successfully induced robust Th1 immune responses in mice and may render pDNA/TMC nanoparticles a potential vaccine candidate for further investigations of protective efficacy against Mtb infections.

Adjuvanted trimethyl chitosan based nanoparticle formulations to improve immunogenicity of DNA vaccines