埃博拉假病毒小鼠攻毒模型的建立与评估  

Establishment and Evaluation of A Mouse Challenge Model Infected A Pseudotyped Ebola Virus

作　　者：孙冰洁 范鹏飞 房婷 于长明 陈薇 SUN Bingjie;FAN Pengfei;FANG Ting;YU Changming;CHEN Wei(Institute of Biotechnology,Academy of Military Medical Sciences(AMMS),Academy of Military Sciences,Beijing 100071,China)

机构地区：[1]军事科学院、军事医学研究院、生物工程研究所,北京100071

出　　处：《病毒学报》2023年第1期213-221,共9页Chinese Journal of Virology

摘　　要：埃博拉病毒具有极大的危险性,对其活病毒相关的操作必须严格限制在生物安全四级实验室(BSL-4)中,阻碍了疫苗和治疗药物的研发进程。本研究将扎伊尔埃博拉病毒(EBOV)膜表面糖蛋白(GP)表达质粒与携带荧光素酶报告基因的pNL4-3.Luc.R-E-慢病毒载体共转染至HEK293T细胞,包装HIV骨架的EBOV GP-Fluc假病毒颗粒,建立了BSL-2条件下基于假病毒和生物发光成像(BLI)技术的BALB/c小鼠EBOV感染模型,并通过不同给药时间、途径和抗体种类的系统评估对其可靠性进行了验证。腹腔注射包含GP全长的假病毒在第4d可检测到集中于胸腺部位的最大发光信号,测试抗体在预防性静脉给药时可完全阻断假病毒的感染,在暴露后腹腔给药时表现出与其在活病毒小鼠模型中一致的保护效力。该模型为埃博拉病毒以及类似烈性病原体防治药物研发提供了一种简单有效的前期筛选方案。Ebolavirus is extremely dangerous, and the operation related to live virus must be strictly restricted in the biosafety level 4 laboratory(BSL-4), thus hindering the development of vaccines and therapeutic drugs. In this study, the Zaire ebolavirus(EBOV) membrane surface glycoprotein(GP) expression plasmid and pNL4-3.Luc. R-E-lentiviral vector carrying the luciferase reporter gene were co-transfected into HEK293T cells to package a human immunodeficiency virus(HIV)-EBOV GP-Luc pseudovirus. A EBOV-challenge model using BALB/c mouse based on pseudoviruses and bioluminescence imaging under BSL-2 facility was established. The reliability of this model was verified by systematic evaluation through different administration timepoints, routes and antibodies. The maximum bioluminescent signal was detected in thymus 4 days post intraperitoneal injection of a pseudovirus containing full-length GP from EBOV. Test antibodies completely blocked pseudovirus infection when administered intravenously as prophylactic agents, and exhibited same protective efficacy when administered intraperitoneally after exposure as in a live virus mouse model. This model provides a simple and effective preliminary screening scheme for development of drugs against virulent pathogens such as Ebola virus.

关 键 词：埃博拉病毒 糖蛋白 假型病毒 小鼠模型 生物发光成像 抗体 

分 类 号：R373.9[医药卫生—病原生物学]