A rapid and quantitative assay for measuring antibody-mediated neutralization of West Nile virus infection
A rapid and quantitative assay for measuring antibody-mediated neutralization of West Nile virus infection
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DOI：
10.1016/j.virol.2005.10.030
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发表时间：
2006-03-01
期刊：
影响因子：
3.7
通讯作者：
Doms, RW
中科院分区：
文献类型：
--
作者：
Pierson, TC;S치nchez, MD;Doms, RW
西尼罗河病毒（West Nile virus，WNV）是一种嗜神经性黄病毒，属于日本脑炎抗原复合物，可引起人类西尼罗河脑炎。WNV病毒粒子的表面被高度有序的二十面体阵列的包膜蛋白覆盖，其负责介导与靶细胞的附着和融合。这些包膜蛋白也是体内产生中和抗体的主要靶标。在这项研究中，我们描述了一种新的方法来测量抗体介导的中和西尼罗河病毒感染的病毒样颗粒，测量感染作为报告基因表达的函数。这些报告病毒颗粒（RVP）通过使用常规DNA表达载体将亚基因组复制子与反式提供的WNV结构蛋白互补来产生。这种方法的精确度和准确度取决于在满足应用于病毒中和的质量作用定律假设的条件下测量抗体和病毒抗原之间相互作用的结果的能力。除了其定量优势外，这种方法还允许产生具有不同WNV毒株和突变体的prM-E蛋白的WNV RVP，为体外研究对WNV的体液免疫应答提供了相当大的灵活性。WNV RVP仅能进行单轮感染，可在BSL-2条件下使用，并提供了一种快速定量检测病毒进入和中和抗体抑制的方法。爱思唯尔公司出版
West Nile virus (WNV) is a neurotropic flavivirus within the Japanese encephalitis antigenic complex that is responsible for causing West Nile encephalitis in humans. The surface of WNV virions is covered by a highly ordered icosahedral array of envelope proteins that is responsible for mediating attachment and fusion with target cells. These envelope proteins are also primary targets for the generation of neutralizing antibodies in vivo. In this study, we describe a novel approach for measuring antibody-mediated neutralization of WNV infection using Virus-like particles that measure infection as a function of reporter gene expression. These reporter virus particles (RVPs) are produced by complementation of a subgenomic replicon with WNV structural proteins provided in trans using conventional DNA expression vectors. The precision and accuracy of this approach stern from an ability to measure the Outcome of the interaction between antibody and viral antigens under conditions that satisfy the assumptions of the law of mass action as applied to virus neutralization. In addition to its quantitative strengths, this approach allows the production of WNV RVPs bearing the prM-E proteins of different WNV strains and mutants, offering considerable flexibility for the study of the humoral immune response to WNV in vitro. WNV RVPs are capable of only a Single round of infection, can be used under BSL-2 conditions, and offer a rapid and quantitative approach for detecting Virus entry and its inhibition by neutralizing antibody. Published by Elsevier Inc.