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人工核酶对非包膜 RNA 病毒的灭活:以急性麻痹病病毒和蜜蜂为实验模型评估抗病毒的体内活性。

Inactivation of a non-enveloped RNA virus by artificial ribonucleases: honey bees and acute bee paralysis virus as a new experimental model for in vivo antiviral activity assessment.

机构信息

Institute of Chemical Biology and Fundamental Medicine, SB RAS, 630090 Novosibirsk, Russian Federation.

出版信息

Antiviral Res. 2011 Sep;91(3):267-77. doi: 10.1016/j.antiviral.2011.06.011. Epub 2011 Jun 22.

Abstract

RNA-containing viruses represent a global threat to the health and wellbeing of humans and animals. Hence, the discovery of new approaches for the design of novel vaccines and antiviral compounds attains high attention. Here we describe the potential of artificial ribonucleases (aRNases), low molecular weight compounds capable to cleave phosphodiester bonds in RNA under mild conditions, to act as antiviral compounds via destroying the genome of non-enveloped RNA viruses, and the potential of utilizing honey bee larvae and adult bees (Apis mellifera) as a novel experimental system for the screening of new antiviral compounds. Pre-incubation of an Acute bee paralysis virus (ABPV) suspension with aRNases D3-12, K-D-1 or Dp12F6 in a concentration-dependent manner increased the survival rate of bee larvae and adult bees subsequently infected with these preparations, whereas incubation of the virus with aRNases ABL3C3 or L2-3 had no effect at all. The results of RT-PCR analysis of viral RNA isolated from aRNase-treated virus particles confirmed that virus inactivation occurs via degradation of viral genomic RNA: dose-dependent inactivation of ABPV correlates well with the cleavage of viral RNA. Electron microscopy analysis revealed that the morphology of ABPV particles inactivated by aRNases remains unaffected as compared to control virus preparations. Altogether the obtained results clearly demonstrate the potential of aRNases as a new virus inactivation agents and bee larvae/ABPV as a new in vivo system for the screening of antiviral compounds.

摘要

含 RNA 的病毒对人类和动物的健康和福祉构成全球性威胁。因此,发现设计新型疫苗和抗病毒化合物的新方法受到高度关注。在这里,我们描述了人工核糖核酸酶 (aRNase) 的潜力,即能够在温和条件下切割 RNA 磷酸二酯键的小分子化合物,可通过破坏非包膜 RNA 病毒的基因组来作为抗病毒化合物发挥作用,以及利用蜜蜂幼虫和成年蜜蜂 (Apis mellifera) 作为筛选新型抗病毒化合物的新型实验系统的潜力。在浓度依赖性方式下,将急性蜜蜂麻痹病毒 (ABPV) 悬浮液与 aRNase D3-12、K-D-1 或 Dp12F6 预孵育可提高随后用这些制剂感染的蜜蜂幼虫和成年蜜蜂的存活率,而将病毒与 aRNase ABL3C3 或 L2-3 孵育则根本没有效果。从用 aRNase 处理的病毒颗粒中分离出的病毒 RNA 的 RT-PCR 分析结果证实,病毒失活是通过降解病毒基因组 RNA 发生的:ABPV 的剂量依赖性失活与病毒 RNA 的切割密切相关。电子显微镜分析显示,与对照病毒制剂相比,aRNase 失活的 ABPV 颗粒的形态保持不变。总的来说,这些结果清楚地表明了 aRNase 作为新型病毒灭活剂的潜力,以及蜜蜂幼虫/ABPV 作为筛选抗病毒化合物的新型体内系统的潜力。

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