Abstract：
Highly pathogenic Arenaviruses, like the Lassa Virus (LASV), pose a serious public health threat in affected countries. Research and development of vaccines and therapeutics are urgently needed but hampered by the necessity to handle these pathogens under biosafety level 4 conditions. These containment restrictions make large-scale screens of antiviral compounds difficult. Therefore, the Mopeia virus (MOPV), closely related to LASV, is often used as an apathogenic surrogate virus. We established for the first time trisegmented MOPVs (r3MOPV) with duplicated S segments, in which one of the viral genes was replaced by the reporter genes ZsGreen (ZsG) or Renilla Luciferase (Rluc), respectively. In vitro characterization of the two trisegmented viruses (r3MOPV ZsG/Rluc and r3MOPV Rluc/ZsG), showed comparable growth behavior to the wild type virus and the expression of the reporter genes correlated well with viral titer. We used the reporter viruses in a proof-of-principle in vitro study to evaluate the antiviral activity of two well characterized drugs. IC50 values obtained by Rluc measurement were similar to those obtained by virus titers. ZsG expression was also suitable to evaluate antiviral effects. The trisegmented MOPVs described here provide a versatile and valuable basis for rapid high throughput screening of broadly reactive antiviral compounds against arenaviruses under BSL-2 conditions.
摘要：
高致病性Arenavirus,比如拉沙病毒(LASV),在受影响的国家构成严重的公共卫生威胁。迫切需要疫苗和治疗剂的研究和开发，但由于在生物安全4级条件下处理这些病原体的必要性而受到阻碍。这些限制使抗病毒化合物的大规模筛选变得困难。因此,莫皮亚病毒（MOPV），与LASV密切相关，通常被用作一种不致病的替代病毒。我们首次建立了具有重复S段的三分段MOPV(r3MOPV)，其中一个病毒基因被报道基因ZsGreen（ZsG）或Renilla荧光素酶（Rluc）取代，分别。两种三段病毒（r3MOPVZsG/Rluc和r3MOPVRluc/ZsG）的体外表征，显示出与野生型病毒相当的生长行为，报告基因的表达与病毒滴度密切相关。我们在原理验证的体外研究中使用了报告病毒，以评估两种特征明确的药物的抗病毒活性。通过Rluc测量获得的IC50值与通过病毒滴度获得的IC50值相似。ZsG表达也适于评估抗病毒作用。这里描述的三分段MOPV为在BSL-2条件下快速高通量筛选针对沙粒病毒的广泛反应性抗病毒化合物提供了通用和有价值的基础。
