Wang Dong Mei, Zhu Jian Bo, Peng Ming, Zhou Peng
State Key Laboratory of Tropical Crop Biotechnology, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Science, Haikou 571101, China.
Transgenic Res. 2008 Dec;17(6):1163-70. doi: 10.1007/s11248-008-9188-1. Epub 2008 Jul 24.
The expression of antigens in transgenic plants has increasingly been used as an alternative to the classical methodologies for the development of experimental vaccines, and it remains one of the real challenges in this field to use transgenic plant-based vaccines effectively as feedstuff additives. We report herein the development of a new oral immunization system for foot and mouth disease with the structural protein VP1 of the foot and mouth disease virus (FMDV) produced in transgenic Stylosanthes guianensis cv. Reyan II. The transgenic plantlets were identified by polymerase chain reaction (PCR), Southern blotting, and northern blotting; and the production of VP1 protein in transgenic plants was confirmed and quantified by western blotting and enzyme-linked immunosorbent assays (ELISA). Six transformed lines were obtained, and the level of the expressed protein was 0.1-0.5% total soluble protein (TSP). Mice that were orally immunized using studded feedstuff mixed with desiccated powder of the transgenic plants developed a virus-specific immune response to the structural VP1 and intact FMDV particles. To our knowledge, this is the first report of transgenic plants expressing the antigen protein of FMDV as feedstuff additives that has demonstrated the induction of a protective systemic antibody response in animals. These results support the feasibility of producing edible vaccines from transgenic forage plants, and provide proof of the possibility of using plant-based vaccines as feedstuff additives.
转基因植物中抗原的表达越来越多地被用作开发实验性疫苗的传统方法的替代方法,而将基于转基因植物的疫苗有效地用作饲料添加剂仍然是该领域的实际挑战之一。我们在此报告了一种新的口蹄疫口服免疫系统的开发,该系统利用在转基因柱花草品种热研2号中产生的口蹄疫病毒(FMDV)结构蛋白VP1。通过聚合酶链反应(PCR)、Southern印迹和Northern印迹鉴定转基因植株;通过Western印迹和酶联免疫吸附测定(ELISA)确认并定量转基因植物中VP1蛋白的产生。获得了6个转化株系,表达蛋白水平为总可溶性蛋白(TSP)的0.1-0.5%。用与转基因植物干燥粉末混合的带钉饲料口服免疫的小鼠对结构VP1和完整FMDV颗粒产生了病毒特异性免疫反应。据我们所知,这是首次报道将表达FMDV抗原蛋白的转基因植物用作饲料添加剂并在动物中诱导出保护性全身抗体反应。这些结果支持了从转基因饲料植物生产可食用疫苗的可行性,并提供了将基于植物的疫苗用作饲料添加剂的可能性的证据。