Santi Luca, Batchelor Lance, Huang Zhong, Hjelm Brooke, Kilbourne Jacquelyn, Arntzen Charles J, Chen Qiang, Mason Hugh S
The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ 85287-5401, USA.
Vaccine. 2008 Mar 28;26(15):1846-54. doi: 10.1016/j.vaccine.2008.01.053. Epub 2008 Feb 15.
Virus-like particles (VLPs) derived from enteric pathogens like Norwalk virus (NV) are well suited to study oral immunization. We previously described stable transgenic plants that accumulate recombinant NV-like particles (rNVs) that were orally immunogenic in mice and humans. The transgenic approach suffers from long generation time and modest level of antigen accumulation. We now overcome these constraints with an efficient tobacco mosaic virus (TMV)-derived transient expression system using leaves of Nicotiana benthamiana. We produced properly assembled rNV at 0.8 mg/g leaf 12 days post-infection (dpi). Oral immunization of CD1 mice with 100 or 250 microg/dose of partially purified rNV elicited systemic and mucosal immune responses. We conclude that the plant viral transient expression system provides a robust research tool to generate abundant quantities of rNV as enriched, concentrated VLP preparations that are orally immunogenic.
源自肠道病原体如诺沃克病毒（NV）的病毒样颗粒（VLP）非常适合用于研究口服免疫。我们之前描述了稳定的转基因植物，其能积累重组NV样颗粒（rNV），这些颗粒在小鼠和人类中具有口服免疫原性。转基因方法存在生成时间长和抗原积累水平中等的问题。我们现在通过使用本氏烟草叶片的高效烟草花叶病毒（TMV）衍生的瞬时表达系统克服了这些限制。在感染后12天（dpi），我们在每克叶片中产生了0.8毫克正确组装的rNV。用100或250微克/剂量的部分纯化rNV对CD1小鼠进行口服免疫可引发全身和黏膜免疫反应。我们得出结论，植物病毒瞬时表达系统提供了一种强大的研究工具，可生成大量富集、浓缩的VLP制剂形式的rNV，这些制剂具有口服免疫原性。
