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重组寨卡 NS1 蛋白从 Vero 细胞中分泌,有利于诱导产生针对 NS1 二聚体的免疫血清。

Recombinant Zika NS1 Protein Secreted from Vero Cells Is Efficient for Inducing Production of Immune Serum Directed against NS1 Dimer.

机构信息

Processus Infectieux en Milieu Insulaire Tropical (PIMIT), Université de La Réunion, INSERM UMR 1187, CNRS 9192, IRD 249, Platform CYROI, 2 rue Maxime Rivière, 97491 Sainte-Clotilde, La Réunion, France.

Section des Sciences Pharmaceutiques, Université de Genève, rue Michel-Servet 1, 1211 Genève, Switzerland.

出版信息

Int J Mol Sci. 2017 Dec 23;19(1):38. doi: 10.3390/ijms19010038.

DOI:10.3390/ijms19010038
PMID:29295477
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5795988/
Abstract

Zika virus (ZIKV) is a mosquito-borne flavivirus that recently emerged in the South Pacific, Americas, and Caribbean islands, where the larger epidemics were documented. ZIKV infection in humans is responsible for neurological disorders and microcephaly. Flavivirus NS1 is a non-structural glycoprotein that is expressed on the cell surface and secreted as a hexameric lipoprotein particle. Intracellular NS1 exists as a dimer that is required for viral replication, whereas the secreted NS1 hexamer interacts with host factors, leading to pathophysiological conditions. In an effort to dispose of specific anti-ZIKV NS1 immune serum, Vero cells were transduced with a lentiviral vector containing the NS1 gene from an epidemic strain of ZIKV. We showed that stably transduced Vero/ZIKV NS1 cell clone was efficient in the secretion of recombinant NS1 oligomer. Immunization of adult rat with purified extracellular NS1 developed anti-ZIKV antibodies that specifically react with the NS1 dimer produced in human cells infected with African and Asian strains of ZIKV. The rat antibody against ZIKV NS1 dimer is a reliable biological tool that enables the immunological detection of secreted NS1 from host-cells infected with ZIKV.

摘要

Zika 病毒(ZIKV)是一种蚊媒黄病毒,最近在南太平洋、美洲和加勒比岛屿出现,这些地区记录了更大规模的流行。人类感染 Zika 病毒会导致神经紊乱和小头症。黄病毒 NS1 是一种非结构糖蛋白,在细胞表面表达并作为六聚体脂蛋白颗粒分泌。细胞内的 NS1 以二聚体形式存在,这是病毒复制所必需的,而分泌的 NS1 六聚体与宿主因子相互作用,导致病理生理状况。为了清除特定的抗-ZIKV NS1 免疫血清,我们用含有来自 Zika 病毒流行株的 NS1 基因的慢病毒载体转导 Vero 细胞。我们表明,稳定转导的 Vero/ZIKV NS1 细胞克隆能够有效地分泌重组 NS1 寡聚物。用纯化的细胞外 NS1 免疫成年大鼠,可产生抗-ZIKV NS1 二聚体的抗体,该抗体特异性地与感染了非洲和亚洲 ZIKV 株的人类细胞中产生的 NS1 二聚体反应。针对 ZIKV NS1 二聚体的大鼠抗体是一种可靠的生物学工具,可用于免疫检测感染了 ZIKV 的宿主细胞中分泌的 NS1。

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