REDBIO México 2010

Folio: 123
Tipo de presentación: Cartel
Tema: Biotecnología Pecuaria

Rotavirus VP8* antigen produced in tobacco transplantomic plants confers protection against challenge in a suckling mouse model.

Lentz Ezequiel M# (1), Mozgovoj Marina V # * (2), Bellido Demian (2), Dus Santos María José (2), Wigdorovitz Andrés (2), BravoAlmonacid Fernando F (1).
* Autor para correspondencia (Corresponding author) awigdorovitz@cnia.inta.gov.ar

1- Laboratorio de Virología y Biotecnología Vegetal, INGEBI-UBA/CONICET, Ciudad Autónoma de Buenos Aires, Vuelta de Obligado 2490, C1428ADN; 2- Instituto de Virología S. Rivenson, Centro de Investigaciones en Ciencias Veterinarias, INTA Castelar, Buenos Aires, Argentina

RESUMEN (ABSTRACT)
"Many plant-based gene expression systems have been developed for the production of recombinant proteins, with the objective of generating cost effective and safe alternatives to conventional production systems. In this context, chloroplast transgenic approach offers several unique advantages over nuclear transformation, which includes, among others, high expression levels of the desired antigen and reduced risk of unwanted transgene spread via pollen and thus increased biosafety of genetically modified plants. Rotavirus VP8* subunit is the minor trypsin cleavage product of the spike protein VP4, which is the major determinant of the viral infectivity and one of the neutralizing antigens. In this work, the C486 bovine rotavirus (BRV) VP8* protein was produced in tobacco chloroplasts. Tobacco transplastomic plants were obtained by particle bombardment and characterized at the molecular level by southern blot, northern blot and western blot. The VP8* expression levels were estimated as 4% of the total soluble proteins or 250 µg per gram of fresh leaf tissue. The VP8* levels remained stable independently from the leaf age. This antigen was able to induce a strong immune response in female mice as measured by ELISA and virus neutralization test. Most importantly, suckling mice born to immunized dams were fully protected against oral challenge with virulent rotavirus. Results presented here demonstrate the feasibility of using antigens expressed in transplantomic plants for the development of subunit vaccines. # Authors contributed equally to the results presented in this report."