Influenza virus-like particles produced by transient expression in Nicotiana benthamiana induce a protective immune response against a lethal viral challenge in mice
Influenza virus-like particles produced by transient expression in Nicotiana benthamiana induce a protective immune response against a lethal viral challenge in mice
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DOI：
10.1111/j.1467-7652.2008.00384.x
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发表时间：
2008-12-01
影响因子：
13.8
通讯作者：
Vezina, Louis-P.
中科院分区：
文献类型：
--
作者：
D'Aoust, Marc-Andre;Lavoie, Pierre-Olivier;Vezina, Louis-P.
株特异性疫苗是应对流感大流行威胁的最佳可能对策。以目前的流感疫苗生产能力，在第一波感染高峰到来之前将这种疫苗快速交付给世界人口似乎是一个无法实现的目标。基于植物的瞬时表达是少数几个可以满足预期激增要求的生产系统之一。为了评估植物农杆菌渗入产生流感疫苗的能力，我们通过本氏烟草植物的农杆菌渗入表达来自菌株A/印度尼西亚/5/05（H5 N1）和A/新喀里多尼亚/20/99（H1N1）的血凝素（HA）。渗透叶中的蛋白质含量的大小分布分析表明，HA主要组装成高分子量的结构。纯化含H5的结构，并通过透射电子显微镜检查确认病毒样颗粒（VLP）组装。VLP脂质组成的高效薄层色谱分析突出了极性和中性脂质含量与来自烟草植物的纯化质膜的含量相当。电镜观察了N.本萨米亚纳叶子证实了VLP在质膜的质外体凹痕中积累。最后，用两个剂量的低至0.1 μ g的纯化流感H5-VLP免疫小鼠引发了针对同源病毒的强烈免疫应答，而两个剂量的0.5 μ g的H5-VLP赋予了针对异源A/Vietnam/1194/04（H5 N1）毒株的致死性攻击的完全保护。这些结果首次表明，植物能够产生从质膜出芽的包膜流感VLP;这种VLP代表了针对流感大流行毒株的疫苗接种的非常有希望的候选物。
A strain-specific vaccine represents the best possible response to the threat of an influenza pandemic. Rapid delivery of such a vaccine to the world's population before the peak of the first infection wave seems to be an unattainable goal with the current influenza vaccine manufacturing capacity. Plant-based transient expression is one of the few production systems that can meet the anticipated surge requirement. To assess the capability of plant agroinfiltration to produce an influenza vaccine, we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants. Size distribution analysis of protein content in infiltrated leaves revealed that HA was predominantly assembled into high-molecular-weight structures. H5-containing structures were purified and examination by transmission electron microscopy confirmed virus-like particle (VLP) assembly. High-performance thin layer chromatography analysis of VLP lipid composition highlighted polar and neutral lipid contents comparable with those of purified plasma membranes from tobacco plants. Electron microscopy of VLP-producing cells in N. benthamiana leaves confirmed that VLPs accumulated in apoplastic indentations of the plasma membrane. Finally, immunization of mice with two doses of as little as 0.1 mu g of purified influenza H5-VLPs triggered a strong immune response against the homologous virus, whereas two doses of 0.5 mu g of H5-VLPs conferred complete protection against a lethal challenge with the heterologous A/Vietnam/1194/04 (H5N1) strain. These results show, for the first time, that plants are capable of producing enveloped influenza VLPs budding from the plasma membrane; such VLPs represent very promising candidates for vaccination against influenza pandemic strains.