GIULIETTI Ana Maria
congresos y reuniones científicas
Expression of a dengue virus antigen and Hepatitis B core fusion protein in N. benthamiana plants
MARTINEZ C;; GIULIETTI AM; RODRÍGUEZ TALOU J; MASSON H
Congreso; XLVI Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2011
Hepatitis B core antigen (HBcore) can self-assemble into VLP (virus-like particles) when it is produced in plants and it has shown to be an attractive carrier for presentation of foreign epitopes for the development of subunit vaccines. The small and critical dengue virus domain III of envelope protein (DVd3) contains multiple serotype-speciﬁc neutralizing epitopes. However, previous work showed that mice immunized with plant-made DVd3 protein without adjuvant elicited low immune responses. In order to produce a candidate for subunit vaccines and to avoid low immune responses in the corresponding pre-clinical studies, DVd3 was fused to HBcore and introduced into tobacco plant cells using deconstructed viral vectors. DVd3 and HBcore antigens were linked by a ﬂexible linker peptide and co-infiltrated in Nicotiana benthamiana plants with 5 Module and Integrase Module. HBcore-DVd3 was produced successfully in N. benthamiana plants (0.43 mg/g fresh weight or 2.23% of total soluble protein) and the fusion protein obtained was reactive with anti-E and anti-HBcore antibodies. Preliminary sedimentation analysis on sucrose gradient showed a particulate nature of the plant-produced fusion protein. Our study demonstrated the potential use of plant-produced HBcore as a carrier protein for immunogenic presentation of dengue epitopes. Electron microscopy studies are currently being done.