Construction of a high-yield dengue virus by replacing nonstructural proteins 3-4B without increasing virulence
Construction of a high-yield dengue virus by replacing nonstructural proteins 3-4B without increasing virulence
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DOI：
10.1016/j.bbrc.2017.11.137
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发表时间：
2018-01-01
影响因子：
3.1
通讯作者：
Kurosu, Takeshi
中科院分区：
文献类型：
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作者：
Phanthanawiboon, Supranee;Pambudi, Sabar;Kurosu, Takeshi
高产量生产病毒对于研制全病毒灭活疫苗或减毒活疫苗至关重要。然而，大多数登革热病毒(DENV)临床分离株在培养细胞中的复制水平较低，这限制了它们用于疫苗开发。本研究检测了低复制DENV临床分离株和高复制实验室分离株之间的差异，目的是设计出高产量的DENV临床分离株。从高复制的实验室4型DENV(DENV-4)H_241株和临床分离株构建的一系列重组嵌合病毒的构建表明，H_241的NS3-NS4B区在培养细胞中具有复制优势。此外，Northern杂交分析表明，这一优势是由于更有效地合成病毒RNA所致。重要的是，替换H241的NS3-NS4B区域并没有增加小鼠的毒力，这表明可以安全地增加病毒产量。这项研究提供了有助于设计可用于疫苗开发的安全和高产病毒的信息。(C)2017 Elsevier Inc.保留所有权利。
Producing virus at high yield is critically important for development of whole virion inactivated vaccines or live attenuated vaccines. Most dengue virus (DENV) clinical isolates, however, replicate at low levels in cultured cells, which limits their use for vaccine development. The present study examined differences between low-replicating DENV clinical isolates and high-replicating laboratory strains with the aim of engineering high-yield DENV clinical isolates. Construction of a series of recombinant chimeric viruses derived from a high-replicating laboratory DENV type 4 (DENV-4) H241 strain and a clinical isolate revealed that the NS3-NS4B region of H241 conferred a replication advantage in cultured cells. Furthermore, northern blot analysis revealed that this advantage was due to more efficient synthesis of viral RNA. Importantly, replacement of the NS3-NS4B region of H241 did not increase virulence in mice, suggesting that viral production can be increased safely. This study provided information that will facilitate engineering of safe and high-yield viruses that can be used for vaccine development. (C) 2017 Elsevier Inc. All rights reserved.