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开发一种重组报告性基孔肯雅病毒用于抗病毒药物筛选检测。

Development of a recombinant reporter Getah virus for antiviral drug screening assays.

机构信息

Laboratory of Animal infectious Diseases and molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning 530005, China.

Laboratory of Animal infectious Diseases and molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning 530005, China; Guangxi Zhuang Autonomous Region Engineering Research Center of Veterinary Biologics, Nanning 530005, China; Guangxi Key Laboratory of Animal Reproduction, Breeding and Disease Control, Nanning 530005, China; Guangxi Colleges and Universities Key Laboratory of Prevention and Control for Animal Disease, Nanning 530005, China.

出版信息

Vet Microbiol. 2023 Jun;281:109742. doi: 10.1016/j.vetmic.2023.109742. Epub 2023 Apr 13.

Abstract

Getah virus (GETV), is an often neglected and re-emerging mosquito-borne RNA virus. GETV can cause illness accompanied with high fever, rash, incapacitating arthralgia and chronic arthritis or encephalitic disease in affected animals. Currently, there is no specific treatment or vaccine against GETV infection. In this study, we developed three recombinant viruses by inserting different reporter protein genes between the Cap and pE2 genes. The reporter viruses exhibited high replication capacity similar to the parental virus. The rGECiLOV and rGECGFP viruses were genetically stable within at least ten rounds of passages in BHK-21 cells. We confirmed that the reporter virus, rGECGFP, facilitated the antiviral assays against GETV by testing it with the known inhibitor, ribavirin. It was also found that the compound, doxycycline, showed an inhibitory effect on GETV replication. In addition, rGECGFP was found to be an authentic mimic of the parental virus infection in 3-day-old mice, but with milder pathogenicity. The reporter viruses will contribute to the assessment of viral replication and proliferation, tracking and elucidating of alphavirus-host interactions. In addition, they will help in the screening of potential antiviral compounds.

摘要

基孔肯雅病毒(GETV)是一种常被忽视但又重新出现的蚊媒 RNA 病毒。该病毒可引起疾病,伴有高热、皮疹、使人丧失能力的关节炎和慢性关节炎或脑炎疾病。目前,尚无针对 GETV 感染的特定治疗或疫苗。在这项研究中,我们通过在 Cap 和 pE2 基因之间插入不同的报告蛋白基因,开发了三种重组病毒。报告病毒表现出与亲本病毒相似的高复制能力。rGECiLOV 和 rGECGFP 病毒在 BHK-21 细胞中至少传代十次仍具有遗传稳定性。我们通过用已知抑制剂利巴韦林测试 rGECGFP 报告病毒,证实了该报告病毒有助于针对 GETV 的抗病毒测定。还发现化合物强力霉素对 GETV 复制具有抑制作用。此外,rGECGFP 在 3 日龄小鼠中被发现是亲本病毒感染的真实模拟物,但致病性较弱。报告病毒将有助于评估病毒复制和增殖、跟踪和阐明甲病毒-宿主相互作用。此外,它们还有助于筛选潜在的抗病毒化合物。

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