基于携带 M2 蛋白胞外结构域的病毒样 HBc 颗粒的植物源性重组流感疫苗。
Plant-produced recombinant influenza vaccine based on virus-like HBc particles carrying an extracellular domain of M2 protein.
机构信息
Centre "Bioengineering", Russian Academy of Sciences, Moscow, Russia.
出版信息
Biochemistry (Mosc). 2012 Jan;77(1):33-40. doi: 10.1134/S000629791201004X.
Conventional influenza vaccines are based on a virus obtained in chicken embryos or its components. The high variability of the surface proteins of influenza virus, hemagglutinin and neuraminidase, requires strain-specific vaccines matching the antigenic specificity of newly emerging virus strains to be developed. A recombinant vaccine based on a highly conservative influenza virus protein M2 fused to a nanosized carrier particle can be an attractive alternative to traditional vaccines. We have constructed a recombinant viral vector based on potato X virus that provides for expression in the Nicotiana benthamiana plants of a hybrid protein M2eHBc consisting of an extracellular domain of influenza virus M2 protein (M2e) fused to hepatitis B core antigen (HBc). This vector was introduced into plant cells by infiltrating leaves with agrobacteria carrying the viral vector. The hybrid protein M2eHBc was synthesized in the infected N. benthamiana plants in an amount reaching 1-2% of the total soluble protein and formed virus-like particles with the M2e peptide presented on the surface. Methods of isolation and purification of M2eHBc particles from plant producers were elaborated. Experiments on mice have shown a high immunogenicity of the plant-produced M2eHBc particles and their protective effect against lethal influenza challenge. The developed transient expression system can be used for production of M2e-based candidate influenza vaccine in plants.
传统的流感疫苗是基于在鸡胚中获得的病毒或其成分。流感病毒表面蛋白血凝素和神经氨酸酶的高度变异性,需要开发与新出现的病毒株的抗原特异性相匹配的株特异性疫苗。一种基于高度保守的流感病毒蛋白 M2 与纳米载体颗粒融合的重组疫苗,可以作为传统疫苗的有吸引力的替代品。我们构建了一种基于马铃薯 X 病毒的重组病毒载体,该载体能够在 Nicotiana benthamiana 植物中表达由流感病毒 M2 蛋白的细胞外结构域(M2e)与乙型肝炎核心抗原(HBc)融合而成的混合蛋白 M2eHBc。该载体通过携带病毒载体的农杆菌浸润叶片而被引入植物细胞。在感染的 N. benthamiana 植物中,M2eHBc 混合蛋白的合成量达到总可溶性蛋白的 1-2%,并形成带有表面呈现的 M2e 肽的病毒样颗粒。从植物生产中分离和纯化 M2eHBc 颗粒的方法已经得到了阐述。在小鼠中的实验表明,植物产生的 M2eHBc 颗粒具有高度的免疫原性,并能对致死性流感攻击提供保护作用。所开发的瞬时表达系统可用于在植物中生产基于 M2e 的候选流感疫苗。