Mass Production of Virus-Like Particles Using Chloroplast Genetic Engineering for Highly Immunogenic Oral Vaccine Against Fish Disease.
Mass Production of Virus-Like Particles Using Chloroplast Genetic Engineering for Highly Immunogenic Oral Vaccine Against Fish Disease.
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DOI：
10.3389/fpls.2021.717952
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发表时间：
2021
影响因子：
5.6
通讯作者：
Kawakami H
中科院分区：
文献类型：
--
作者：
Nakahira Y;Mizuno K;Yamashita H;Tsuchikura M;Takeuchi K;Shiina T;Kawakami H
神经坏死病毒（viral nervous necrosis virus，NNV）是引起病毒性神经坏死（viral nervous necrosis，VNN）的病原体，是世界范围内导致鱼类大量死亡的最严重的鱼类疾病之一。虽然有几种注射灭活疫苗可在市场上买到，但仍需要更节省人力、成本效益高和对鱼类友好的免疫方法。利用转基因植物表达病原体来源的重组抗原作为可食用疫苗是满足这些要求的理想途径。本研究利用叶绿体基因工程技术成功地表达了红斑石斑鱼NNV衣壳蛋白（RGNV-CP）。RGNNV-CP在转质体烟草植物的所有幼叶、成熟叶和衰老叶中以高水平积累（平均约3 mg/g叶鲜重）。RGNNV-CP在转基因株系的叶绿体基质中有效地自组装成病毒样颗粒（RGNNV-VLP），这可以很容易地通过原位透射电子显微镜观察到。此外，腹膜内注射和口服给药的粗纯化的蛋白质提取物含有叶绿体衍生的RGNNV-VLP提供了七带石斑鱼对RGNNV的挑战有足够的保护，其免疫原性与商业注射疫苗相当。这些发现表明，叶绿体衍生的VLP疫苗可能在预防各种疾病中发挥有希望的作用，不仅在鱼类，而且在其他动物，包括人类。
Nervous necrosis virus (NNV) is the causative agent of viral nervous necrosis (VNN), which is one of the most serious fish diseases leading to mass mortality in a wide range of fish species worldwide. Although a few injectable inactivated vaccines are commercially available, there is a need for more labor-saving, cost-effective, and fish-friendly immunization methods. The use of transgenic plants expressing pathogen-derived recombinant antigens as edible vaccines is an ideal way to meet these requirements. In this study, chloroplast genetic engineering was successfully utilized to overexpress the red-spotted grouper NNV capsid protein (RGNNV-CP). The RGNNV-CP accumulated at high levels in all young, mature, and old senescent leaves of transplastomic tobacco plants (averaging approximately 3 mg/g leaf fresh weight). The RGNNV-CP efficiently self-assembled into virus-like particles (RGNNV-VLPs) in the chloroplast stroma of the transgenic lines, which could be readily observed by in situ transmission electron microscopy. Furthermore, intraperitoneal injection and oral administration of the crudely purified protein extract containing chloroplast-derived RGNNV-VLPs provided the sevenband grouper fish with sufficient protection against RGNNV challenge, and its immunogenicity was comparable to that of a commercial injectable vaccine. These findings indicate that chloroplast-derived VLP vaccines may play a promising role in the prevention of various diseases, not only in fish but also in other animals, including humans.
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影响因子：
6.3
作者：
Castiglia D;Sannino L;Marcolongo L;Ionata E;Tamburino R;De Stradis A;Cobucci-Ponzano B;Moracci M;La Cara F;Scotti N
通讯作者：
Scotti N
影响因子：
3.3
作者：
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通讯作者：
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影响因子：
1.8
作者：
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通讯作者：
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影响因子：
20.5
作者：
Jin S;Daniell H
通讯作者：
Daniell H
影响因子：
4.7
作者：
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通讯作者：
Kim, Hong-Jin