Virus-like particle expression and assembly in plants: hepatitis B and Norwalk viruses
Virus-like particle expression and assembly in plants: hepatitis B and Norwalk viruses
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DOI：
10.1016/j.vaccine.2004.11.017
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发表时间：
2005-03-07
期刊：
影响因子：
5.5
通讯作者：
Mason, HS
中科院分区：
文献类型：
--
作者：
Huang, Z;Elkin, G;Mason, HS
疫苗抗原在植物中的表达和通过摄入转基因植物材料传递已经在许多临床前动物研究和一些临床试验中显示出希望。已经测试了许多不同的病毒抗原，其中最有希望的是那些可以组装病毒样颗粒(VLP)的病毒样颗粒，这种颗粒模仿真正的病毒粒子的形式，并显示中和抗体表位。我们广泛地研究了乙肝表面抗原(HBs)和诺沃克病毒衣壳蛋白(NVCP)的植物表达、VLP组装和免疫原性。在烟草和大豆悬浮培养细胞以及马铃薯叶片和块茎组织中，观察到乙型肝炎表面抗原小蛋白(S蛋白)可组装来源于内质网的管状膜复合体。这种土豆材料在摄入后对小鼠具有免疫原性。本文描述了乙肝表面抗原中间蛋白(M蛋白或Pre-S2+S)的植物表达和免疫原性，该蛋白在S蛋白的N端增加了55个氨基酸的Pre-S2区域。植物来源的重组M蛋白系统注射小鼠后，比S蛋白诱导更强的抗-HBs抗体应答。我们讨论了融合蛋白在增强免疫原性和粘膜靶向性方面的应用，以及异源融合抗原的传递。NVCP在组装成38 nm病毒粒子大小的二十面体(T=3)VLP的植物中表达，与在昆虫细胞中产生的VLP相似。当小鼠和人摄入新鲜马铃薯块茎时，VLP刺激了它们的血清免疫球蛋白和免疫球蛋白A反应。在这里，我们表明，冷冻干燥的转基因番茄果实产生了稳定的制剂，当喂给小鼠时，这些制剂可以刺激对NVCP的良好的免疫球蛋白和免疫球蛋白A反应。然而，番茄果实中VLP的主要形式是在昆虫细胞来源的NVCP中也观察到了23 nm的小颗粒。(C)2004爱思唯尔有限公司。保留所有权利。
Expression of vaccine antigens in plants and delivery via ingestion of transgenic plant material has shown promise in numerous pre-clinical animal studies and in a few clinical trials. A number of different viral antigens have been tested, and among the most promising are these that can assemble virus-like particles (VLP), which mimic the form of authentic virions and display neutralizing antibody epitopes. We have extensively studied plant expression, VLP assembly, and immunogenicity of hepatitis B surface antigen (HBsAg) and Norwalk virus capsid protein (NVCP). The HBsAg small protein (S protein) was found by TEM to assemble tubular membrane complexes derived from endoplasmic reticulum in suspension cultured cells of tobacco and soybean, and in potato leaf and tuber tissues. The potato material was immunogenic in mice upon delivery by ingestion. Here we describe the plant expression and immunogenicity of HBsAg middle protein (M protein or pre-S2 + S) which contains additional 55 amino acid pre-S2 region at N-terminus of the S protein. Plant-derived recombinant M protein provoked stronger serum antibody responses against HBsAg than did S protein when injected systemically in mice. We discuss implications for use of fusion proteins for enhanced immunogenicity and mucosal targeting of HBsAg, as well as delivery of heterologous fused antigens. NVCP expressed in plants assembled 38 nm virion-size icosahedral (T=3) VLP, similar to those produced in insect cells. The VLP stimulated serum IgG and IgA responses in mice and humans when they were delivered by ingestion of fresh potato tuber. Here we show that freeze-drying of transgenic NVCP tomato fruit yielded stable preparations that stimulated excellent IgG and IgA responses against NVCP when fed to mice. However, the predominant VLP form in tomato fruit was the small 23 nm particle also observed in insect cell-derived NVCP. (c) 2004 Elsevier Ltd. All rights reserved.