Dengue virus (DENV) is an emerging threat causing an estimated 390 million infections per year. Dengvaxia, the only licensed vaccine, may not be adequately safe in young and seronegative patients; hence, development of a safer, more effective vaccine is of great public health interest. Virus‐like particles (VLPs) are a safe and very efficient vaccine strategy, and DENV VLPs have been produced in various expression systems. Here, we describe the production of DENV VLPs in Nicotiana benthamiana using transient expression. The co‐expression of DENV structural proteins (SP) and a truncated version of the non‐structural proteins (NSPs), lacking NS5 that contains the RNA‐dependent RNA polymerase, led to the assembly of DENV VLPs in plants. These VLPs were comparable in appearance and size to VLPs produced in mammalian cells. Contrary to data from other expression systems, expression of the protein complex prM‐E was not successful, and strategies used in other expression systems to improve the VLP yield did not result in increased yields in plants but, rather, increased purification difficulties. Immunogenicity assays in BALB/c mice revealed that plant‐made DENV1‐SP + NSP VLPs led to a higher antibody response in mice compared with DENV‐E domain III displayed inside bluetongue virus core‐like particles and a DENV‐E domain III subunit. These results are consistent with the idea that VLPs could be the optimal approach to creating candidate vaccines against enveloped viruses.

中文翻译：

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由结构蛋白和非结构蛋白共表达产生的植物性登革热病毒样颗粒可诱导小鼠体液免疫反应


登革热病毒 (DENV) 是一种新兴威胁，每年估计造成 3.9 亿人感染。 Dengvaxia 是唯一获得许可的疫苗，对于年轻和血清阴性患者可能不够安全；因此，开发更安全、更有效的疫苗具有重大的公共卫生利益。病毒样颗粒（VLP）是一种安全且非常有效的疫苗策略，并且 DENV VLP 已在各种表达系统中产生。在这里，我们描述了使用瞬时表达在本塞姆氏烟草中产生 DENV VLP。 DENV 结构蛋白 (SP) 和截短版本的非结构蛋白 (NSP) 的共表达，缺乏包含 RNA 依赖性 RNA 聚合酶的 NS5，导致 DENV VLP 在植物中组装。这些 VLP 的外观和大小与哺乳动物细胞中产生的 VLP 相当。与其他表达系统的数据相反，蛋白质复合物 prM-E 的表达并不成功，其他表达系统中使用的提高 VLP 产量的策略并没有导致植物产量增加，而是增加了纯化难度。 BALB/c 小鼠的免疫原性测定表明，与蓝舌病毒核心样颗粒内显示的 DENV-E 结构域 III 和 DENV-E 结构域 III 亚基相比，植物制造的 DENV1-SP + NSP VLP 在小鼠体内产生了更高的抗体反应。这些结果与 VLP 可能是制造针对包膜病毒的候选疫苗的最佳方法的想法是一致的。