Plant virus and plant virus like particles as vaccine components: computational and experimental virology - Laboratory of Biotechnology, Technical Unit Radiation Biology and Human Health, ENEA

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Plant virus and plant virus like particles as vaccine components: computational and experimental virology

Research > Plant Derived Vaccines

Plant virus coat proteins (CP) are particularly suitable carriers to present immunogenic peptides to the immune system. When properly fused at different positions on the capsid proteins, exogenous sequences are expressed in plants, originating recombinant viral CP able to self-assemble and generate chimeric virus particles (CVPs) displaying the foreign sequence on their outer surfaces. The “epitope-displaying” strategy using plant virus CP as carriers for both viral and bacterial antigens has been successfully employed for the production of experimental vaccines. Our research area of expertise, since 1999, is mainly focused in the production of plant chimeric virus particles exposing immunogenic epitopes. In 2001, we provided the evidence that Potato virus X (PVX) derived CVPs, exposing the HIV-1 2F5 neutralizing epitope on their surface, were able to elicit high levels of HIV-1-specific immunoglobulin G (IgG) and IgA antibodies in normal mice without adjuvants. Moreover, hu-PBL-SCID mice immunized with CVP-pulsed autologous dendritic cells (DCs) were able to mount a specific human primary antibody response against the HIV-1-derived epitope. Remarkably, sera obtained from both normal and hu-PBLSCID mice were endowed with anti-HIV-1-neutralizing activity.Afterwards, we have isolated and characterised a mutant PVX clone carrying an N-terminal deleted CP. Currently we are investigating the Tombusvirus Artichoke Mosaic Crinkle virus (AMCV) coat protein as epitope presentation system. We have transiently expressed the sequence encoding the viral CP in Nicotiana benthamiana plants and the assembled virus like particles (VLP) were purified by sucrose gradient and visualised by electron microscopy. Furthermore, we are developing an approach that combines in silico tools and experimental virology for a rational design of immunologically active chimeric VLPs, in an attempt to select suitable sites on the AMCV CP for the insertion of different HIV-1 neutralizing epitopes without affecting VLPs assembly.

Carla Marusic

 
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