Clinical Microbiology: Open Access

Production and optimization of dengue virus- like particles for immunization

4^th International Conference on Clinical Microbiology and Microbial Genomics

October 05-07, 2015 Philadelphia, USA

G Singh, V N Chouljenko, K G Kousoulas and A J Ramsay

LSU Health Sciences Center, USA

Posters-Accepted Abstracts: Clin Microbiol

Abstract:

Dengue virus (DENV) causes more than 100 million cases of dengue fever (DF) annually, making it the common and widespread viral infection of the humans and there are no protective vaccines available currently. Moreover, individuals exposed to DENV infections earlier appear to be more susceptible to complications after secondary infection, a phenomenon attributed to antibodydependent enhancement (ADE). DENV serotypes can be readily produced in cell culture through the expression of the heterologous pre-membrane/membrane (prM/M) and envelope (E) structural proteins that contain the majority of immunogenic domains for antibodies against DENV, have recently gained considerable attention for studies of viral replication and as potential vaccine target. Baculovirus expression systems in insect cells have emerged as a highly efficient means of producing sub-unit vaccines that generate potent protective immune responses against a variety of viral pathogens. We made DENV-2 VLPs through heterologous expression of the full prM/M and E structural proteins in insect cell cultures such as Sf9 and high five cells. The prM/M and E genes were successfully synthesized as codon-optimized gene cassettes in recombinant baculoviruses, expressing combinations of DENV-2 prM/M and E proteins that packaged a DENV-2 replicon with a single poly prM/M-E gene and produced single-round VLP in culture. After recovery of recombinant VLP from cells, expression of DENV-2 proteins was checked by western blotting and later confirmed for VLP production through transmission electron microscopy. Studies are underway to test the efficacy and immunogenicity of these DNA constructs under the appropriate animal models. Vaccine antigens generated through this robust system may serve as a candidate DENV vaccines.

Biography :

Email: gyancdri@gmail.com