The fusion of Toxoplasma gondii SAG1 vaccine candidate to Leishmania infantum heat shock protein 83-kDa improves expression levels in tobacco chloroplasts
| Autor: | Inmaculada Farran, Marina Clemente, Mariana G. Corigliano, María L. Yácono, Sebastián Pariani, Jon Veramendi, Melina Laguía Becher, Edwin F. Sánchez López, Valeria A. Sander, Romina M. Albarracín |
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| Přispěvatelé: | IdAB - Instituto de Agrobiotecnología / Agrobioteknologiako Institutua, Consejo Nacional de Investigaciones Científicas y Técnicas (Argentina), Universidad Nacional de San Martín, Bunge & Born Foundation |
| Rok vydání: | 2015 |
| Předmět: |
Protozoan Vaccines
Chloroplasts Recombinant Fusion Proteins animal diseases Biotecnología Agropecuaria Protozoan Proteins Antibodies Protozoan Toxoplasma gondii Antigens Protozoan Chloroplast transformation Applied Microbiology and Biotechnology Mice Transformation Genetic Antigen Heat shock protein Fusion Protein Tobacco parasitic diseases Animals Humans SAG1 Leishmania infantum Heat-Shock Proteins biology Protein Stability Vaccination food and beverages General Medicine biology.organism_classification Fusion protein Virology Hsp83 Chloroplast Transformation (genetics) Immunoglobulin M CIENCIAS AGRÍCOLAS Immunoglobulin G Biotecnología Agrícola y Biotecnología Alimentaria Molecular Medicine Immunization Female Transplastomic plant |
| Zdroj: | Academica-e: Repositorio Institucional de la Universidad Pública de Navarra Universidad Pública de Navarra Digital.CSIC. Repositorio Institucional del CSIC instname Academica-e. Repositorio Institucional de la Universidad Pública de Navarra |
| Popis: | Special Issue: Vaccine Biotechnology. Chloroplast transformation technology has emerged as an alternative platform offering many advantages over nuclear transformation. SAG1 is the main surface antigen of the intracellular parasite Toxoplasma gondii and a promising candidate to produce an anti-T. gondii vaccine. The aim of this study was to investigate the expression of SAG1 using chloroplast transformation technology in tobacco plants. In order to improve expression in transplastomic plants, we also expressed the 90-kDa heat shock protein of Leishmania infantum (LiHsp83) as a carrier for the SAG1 antigen. SAG1 protein accumulation in transplastomic plants was approximately 0.1-0.2 μg per gram of fresh weight (FW). Fusion of SAG1 to LiHsp83 significantly increased the level of SAG1 accumulation in tobacco chloroplasts (by up to 500-fold). We also evaluated the functionality of the chLiHsp83-SAG1. Three human seropositive samples reacted with SAG1 expressed in transplastomic chLiHsp83-SAG1 plants. Oral immunization with chLiHsp83-SAG1 elicited a significant reduction of the cyst burden that correlated with an increase of SAG1-specific antibodies. We propose the fusion of foreign proteins to LiHsp83 as a novel strategy to increase the expression level of the recombinant proteins using chloroplast transformation technology, thus addressing one of the current challenges for this approach in antigen protein production. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. This work was supported by PIP 0494CO of the National Research Council (CONICET, Argentina), University of General San Martín (UNSAM, Argentina) and Bunge & Born Foundation (Argentina). |
| Databáze: | OpenAIRE |
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