Production of a Plant-Derived Immunogenic Protein Targeting ApoB100 and CETP: Toward a Plant-Based Atherosclerosis Vaccine
Production of a Plant-Derived Immunogenic Protein Targeting ApoB100 and CETP: Toward a Plant-Based Atherosclerosis Vaccine
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DOI：
10.1007/s12033-014-9793-6
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发表时间：
2014-12-01
影响因子：
2.6
通讯作者：
Banuelos-Hernandez, Bernardo
中科院分区：
文献类型：
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作者：
Alberto Salazar-Gonzalez, Jorge;Rosales-Mendoza, Sergio;Banuelos-Hernandez, Bernardo
为了建立一种基于植物的动脉粥样硬化疫苗模型，研究人员设计了一种基于霍乱毒素B亚基（CTB）的嵌合蛋白，以靶向与动脉粥样硬化免疫治疗作用相关的ApoB100和CETP表位。表位在CTB的c端融合产生CTB:p210:CETPe蛋白。合成的CTB:p210:CETPe基因成功转移到烟草植株上，未发生表型改变。植物源性CTB:p210:CETPe以五聚体形式表达和组装。GM1-ELISA和Western blot分析显示，该蛋白保留了目标抗原决定因子。高表达系在叶片组织中的重组蛋白积累水平高达10 μ g/g鲜重，通过qPCR检测，这些系携带一个转基因插入。此外，当皮下给药时，这些CTB:p210: cetpe产生植物的生物量能够在小鼠体内引起针对ApoB100和CETP表位以及人血清蛋白的体液反应。这些发现首次证明了动脉粥样硬化相关的表位可以在植物中表达，保持免疫原性，为动脉粥样硬化疫苗的开发开辟了分子农业领域的新途径。
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 A mu g/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.