Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays.
Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays.
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DOI：
10.3390/v10110655
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发表时间：
2018-11-20
期刊：
影响因子：
--
通讯作者：
Kuhn JH
中科院分区：
文献类型：
--
作者：
Caì Y;Iwasaki M;Beitzel BF;Yú S;Postnikova EN;Cubitt B;DeWald LE;Radoshitzky SR;Bollinger L;Jahrling PB;Palacios GF;de la Torre JC;Kuhn JH
拉萨病毒(LASV)是一种哺乳动物病毒，估计每年在西非感染10万至30万人，并经常导致致命疾病。目前，还没有LASV特异性抗病毒药物或疫苗可用于预防或治疗由LASV引起的拉沙热。医学对抗筛查平台的开发是生产可许可产品的关键一步。我们利用反向遗传学技术构建了重组野生型LASV(rLASV-WT)及其编码可切割绿色荧光蛋白(GFP)的改良型，作为快速定量检测感染的报告基因(rLASV-GFP)。RLASV-WT和野生型LASV在培养细胞中的生长动力学相似，而rLASV-GFP的生长受到轻微的抑制。RLASV-GFP在Vero细胞中连续传代后，GFP报告基因的表达保持稳定。利用两种广谱抗病毒药物法韦拉韦和利巴韦林抑制LASV感染，我们证明rLASV-GFP是鉴定LASV感染抑制剂的一种合适的筛选工具。在这些发现的基础上，我们建立了基于rLASV-GFP的高通量药物发现筛选和基于rLASV-GFP的抗体中和试验。这两个平台现在作为IRF-Frederick(国际资源)的标准工具可用，将加快抗LASV医学对策的发现，并降低最大限度遏制实验室的抗病毒筛查的成本。
Lassa virus (LASV), a mammarenavirus, infects an estimated 100,000–300,000 individuals yearly in western Africa and frequently causes lethal disease. Currently, no LASV-specific antivirals or vaccines are commercially available for prevention or treatment of Lassa fever, the disease caused by LASV. The development of medical countermeasure screening platforms is a crucial step to yield licensable products. Using reverse genetics, we generated a recombinant wild-type LASV (rLASV-WT) and a modified version thereof encoding a cleavable green fluorescent protein (GFP) as a reporter for rapid and quantitative detection of infection (rLASV-GFP). Both rLASV-WT and wild-type LASV exhibited similar growth kinetics in cultured cells, whereas growth of rLASV-GFP was slightly impaired. GFP reporter expression by rLASV-GFP remained stable over several serial passages in Vero cells. Using two well-characterized broad-spectrum antivirals known to inhibit LASV infection, favipiravir and ribavirin, we demonstrate that rLASV-GFP is a suitable screening tool for the identification of LASV infection inhibitors. Building on these findings, we established a rLASV-GFP-based high-throughput drug discovery screen and an rLASV-GFP-based antibody neutralization assay. Both platforms, now available as a standard tool at the IRF-Frederick (an international resource), will accelerate anti-LASV medical countermeasure discovery and reduce costs of antiviral screens in maximum containment laboratories.
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影响因子：
158.5
作者：
MCCORMICK, JB;KING, IJ;BELMONTWILLIAMS, R
通讯作者：
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DOI：
10.4269/ajtmh.1970.19.670
发表时间：
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作者：
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DOI：
10.1093/trstmh/tru009
发表时间：
2014-03-01
影响因子：
2.2
作者：
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通讯作者：
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