Establishment of RNA vaccine techniques using RNA replicons stabilized by cationic liposomes in vitro and in vivo

Abstract

In this project we propose the use of RNA replicons encapsulated in cationic liposomes as a delivery method that should be able to generate in vitro and in vivo therapeutic effects. At first, the cationic liposomes will be analyzed regarding their physicochemical properties such as charge, size and ability to protect the encapsulated RNA. Afterwards, assays will be performed with liposomes varying the concentration of RNA replicons containing the reporter genes gfp and luciferase, in order to obtain the best liposome-RNA replicon ratio for the in vitro essays. For liposome delivery in animals, the experiments will follow the ratios characterized in vitro, which will be used in intradermal immunizations to validate the in vivo data. Lastly, the effect of these immunizations will be compared to the one generated by DNA vaccines. This will be done using an in vivo imaging system (IVIS), with the purpose of obtaining an efficacy comparison between these vaccine models. Once characterized, this system might influence immunization studies using antigens relevant for Plasmodium sp., the causative agent of human malaria.