title: Production of a plant-derived immunogenic protein targeting ApoB100 and CETP: toward a plant-based atherosclerosis vaccine.
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.
authors.affiliation: 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.
authors.forename: Jorge Alberto
authors.initials: JA
authors.lastname: Salazar-Gonzalez
authors.forename: Sergio
authors.initials: S
authors.lastname: Rosales-Mendoza
authors.forename: Andrea
authors.initials: A
authors.lastname: Romero-Maldonado
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Monreal-Escalante
authors.forename: Edith Elena
authors.initials: EE
authors.lastname: Uresti-Rivera
authors.forename: Bernardo
authors.initials: B
authors.lastname: Bañuelos-Hernández
mesh_terms: D000818:Animals; D053299:Apolipoprotein B-100 / Q000276:immunology*; D050197:Atherosclerosis / Q000517:prevention & control; D002772:Cholera Toxin / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D053480:Cholesterol Ester Transfer Proteins / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D051379:Mice; D030821:Plants, Genetically Modified / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D014026:Nicotiana / Q000235:genetics*; D014612:Vaccines / Q000276:immunology
chemical_list: D053299:Apolipoprotein B-100; D053480:Cholesterol Ester Transfer Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D014612:Vaccines; D002772:Cholera Toxin