PDF(Pubmed)
Abstract：
Arboviruses are etiological agents in an extensive group of emerging diseases with great clinical relevance in Brazil, due to the wide distribution of their vectors and the favorable environmental conditions. Among them, the Mayaro virus (MAYV) has drawn attention since its emergence as the etiologic agent of Mayaro fever, a highly debilitating disease. To study viral replication and identify new drug candidates, traditional antiviral assays based on viral antigens and/or plaque assays have been demonstrating low throughput, making it difficult to carry out larger-scale assays. Therefore, we developed and characterized two DNA-launched infectious clones reporter viruses based on the MAYV strain BeAr 20290 containing the reporter genes of firefly luciferase (FLuc) and nanoluciferase (NLuc), designated as MAYV-firefly and MAYV-nanoluc, respectively. The viruses replicated efficiently with similar properties to the parental wild-type MAYV, and luminescence expression levels reflected viral replication. Reporter genes were also preserved during passage in cell culture, remaining stably expressed for one round of passage for MAYV-firefly and three rounds for MAYV-nanoluc. Employing the infectious clone, we described the effect of Rimantadine, an FDA-approved Alzheimer\'s drug, as a repurposing agent for MAYV but with a broad-spectrum activity against Zika virus infection. Additionally, we validated MAYV-nanoluc as a tool for antiviral drug screening using the compound EIDD-2749 (4\'-Fluorouridine), which acts as an inhibitor of alphavirus RNA-dependent RNA polymerase.
摘要：
在巴西，虫媒病毒是一组广泛的新兴疾病的病因，具有极大的临床意义。由于其载体的广泛分布和有利的环境条件。其中,Mayaro病毒（MAYV）自从作为Mayaro热的病原体出现以来就引起了人们的注意，一种非常使人衰弱的疾病。为了研究病毒复制并确定新的候选药物，基于病毒抗原和/或噬斑测定的传统抗病毒测定已显示出低通量，使得难以进行大规模的检测。因此,我们基于MAYV菌株BeAr20290开发并表征了两种DNA启动的感染性克隆报告病毒，其中包含萤火虫荧光素酶（FLuc）和纳米荧光素酶（NLuc）的报告基因，指定为MAYV-萤火虫和MAYV-nanoluc，分别。这些病毒以与亲本野生型MAYV相似的特性有效复制，和发光表达水平反映病毒复制。报告基因在细胞培养物中的传代过程中也被保留，对于MAYV-萤火虫，在一轮传代中保持稳定表达，对于MAYV-nanoluc，在三轮传代中保持稳定表达。利用传染性克隆，我们描述了金刚乙胺的作用,FDA批准的阿尔茨海默氏症药物，作为MAYV的再用途剂，但具有抗寨卡病毒感染的广谱活性。此外,我们验证了MAYV-nanoluc作为使用化合物EIDD-2749（4'-氟尿嘧啶）的抗病毒药物筛选的工具，作为甲病毒RNA依赖性RNA聚合酶的抑制剂。
