Generation of Multiple Arbovirus-like Particles Using a Rapid Recombinant Vaccinia Virus Expression Platform.

Generation of Multiple Arbovirus-like Particles Using a Rapid Recombinant Vaccinia Virus Expression Platform.
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DOI:
10.3390/pathogens11121505
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发表时间:
2022-12-09
期刊:
Pathogens (Basel, Switzerland)
影响因子:
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其他
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正如 2015 年美洲爆发的寨卡病毒所证明的那样,新出现和重新出现的虫媒病毒是公共卫生威胁,需要研究投资来开发有效的预防和治疗方法。许多虫媒病毒疾病未被充分报告、被忽视或流行率较低,但它们都有可能引起当地和国际关注的疫情爆发。在这里,我们展示了使用快速高效的重组痘苗病毒(VACV)表达系统针对五种蜱和蚊媒病毒生产病毒样颗粒(VLP):Powassan病毒(POWV)、Heartland病毒(HRTV)、严重发热伴血小板减少综合征病毒(SFTSV)、波旁病毒(BRBV)和Mayaro病毒(MAYV)。我们通过蛋白质印迹检测了感兴趣的虫媒病毒基因的表达,并在透射电子显微镜下观察了类似于天然病毒粒子的VLP的表达。我们还能够通过修改衣壳基因内的信号序列来改善 POWV VLP 的分泌。本研究描述了使用快速 VACV 平台来生产和纯化可用作亚单位或载体疫苗的虫媒病毒 VLP,并提供了对用于 VLP 形成的虫媒病毒基因的选择和遗传修饰以提高 VLP 分泌和产量的见解。
As demonstrated by the 2015 Zika virus outbreak in the Americas, emerging and re-emerging arboviruses are public health threats that warrant research investment for the development of effective prophylactics and therapeutics. Many arboviral diseases are underreported, neglected, or of low prevalence, yet they all have the potential to cause outbreaks of local and international concern. Here, we show the production of virus-like particles (VLPs) using a rapid and efficient recombinant vaccinia virus (VACV) expression system for five tick- and mosquito-borne arboviruses: Powassan virus (POWV), Heartland virus (HRTV), severe fever with thrombocytopenia syndrome virus (SFTSV), Bourbon virus (BRBV) and Mayaro virus (MAYV). We detected the expression of arbovirus genes of interest by Western blot and observed the expression of VLPs that resemble native virions under transmission electron microscopy. We were also able to improve the secretion of POWV VLPs by modifying the signal sequence within the capsid gene. This study describes the use of a rapid VACV platform for the production and purification of arbovirus VLPs that can be used as subunit or vectored vaccines, and provides insights into the selection of arbovirus genes for VLP formation and genetic modifications to improve VLP secretion and yield.
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