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Production of a Plant-Derived Immunogenic Protein Targeting ApoB100 and CETP: Toward a Plant-Based Atherosclerosis Vaccine

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Abstract

In an effort to initiate the development of a plant-based vaccination model against atherosclerosis, a cholera toxin B subunit (CTB)-based chimeric protein was designed to target both ApoB100 and CETP epitopes associated with immunotherapeutic effects in atherosclerosis. Epitopes were fused at the C-terminus of CTB to yield a protein called CTB:p210:CETPe. A synthetic gene coding for CTB:p210:CETPe was successfully transferred to tobacco plants with no phenotypic alterations. Plant-derived CTB:p210:CETPe was expressed and assembled in the pentameric form. This protein retained the target antigenic determinants, as revealed by GM1-ELISA and Western blot analyses. Higher expresser lines reached recombinant protein accumulation levels up to 10 µg/g fresh weight in leaf tissues and these lines carry a single insertion of the transgene as determined by qPCR. Moreover, when subcutaneously administered, the biomass from these CTB:p210:CETPe-producing plants was able to elicit humoral responses in mice against both ApoB100 and CETP epitopes and human serum proteins. These findings evidenced for the first time that atherosclerosis-related epitopes can be expressed in plants retaining immunogenicity, which opens a new path in the molecular farming field for the development of vaccines against atherosclerosis.

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Acknowledgments

Projects from the group were funded by Grants from CONACYT (102109/56980), PROMEP-2010 to Bioprocess CA. Omar González corrected the English version of the manuscript.

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Authors and Affiliations

  1. Laboratorio de Biofarmacéuticos Recombinantes, Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, 78210, San Luis Potosí, SLP, Mexico

    Jorge Alberto Salazar-Gonzalez, Sergio Rosales-Mendoza, Andrea Romero-Maldonado, Elizabeth Monreal-Escalante & Bernardo Bañuelos-Hernández

  2. Laboratorio de Inmunotoxicología, Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, 78210, San Luis Potosí, SLP, Mexico

    Edith Elena Uresti-Rivera

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  1. Jorge Alberto Salazar-Gonzalez
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Correspondence to Sergio Rosales-Mendoza.

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Jorge Alberto Salazar-Gonzalez and Sergio Rosales-Mendoza have contributed equally to this work.

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Salazar-Gonzalez, J.A., Rosales-Mendoza, S., Romero-Maldonado, A. et al. Production of a Plant-Derived Immunogenic Protein Targeting ApoB100 and CETP: Toward a Plant-Based Atherosclerosis Vaccine. Mol Biotechnol 56, 1133–1142 (2014). https://doi.org/10.1007/s12033-014-9793-6

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