Development of cell-based pseudovirus entry assay to identify potential viral entry inhibitors and neutralizing antibodies against SARS-CoV-2  

作　　者：Jie Hu Qingzhu Gao Changlong He Ailong Huang Ni Tang Kai Wang 

机构地区：[1]Key Laboratory of Molecular Biology for Infectious Diseases(Ministry of Education),Institute for Viral Hepatitis,Department of Infectious Diseases,The Second Affiliated Hospital,Chongqing Medical University,Chongqing,400010,China

出　　处：《Genes & Diseases》2020年第4期551-557,共7页基因与疾病（英文）

基　　金：This work was supported by the Emergency Project from the Science&Technology Commission of Chongqing(cstc2020jscx-fyzx0053);a Major National Science&Technology Program grant(2017ZX10202203)from the Science&Technology Commission of China,the Leading Talent Program of CQ CSTC(CSTCCXLJRC201719);the Scientific Research Innovation Project for Postgraduate in Chongqing(CYB19168);the Emergency Project for Novel Coronavirus Pneumonia from the Chongqing Medical University(CQMUNCP0302).

摘　　要：Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)is the causative virus of the coronavirus disease 2019(COVID-19)pandemic.To establish a safe and convenient assay system for studying entry inhibitors and neutralizing antibodies against SARS-CoV-2,we constructed a codon-optimized,full-length C-terminal mutant spike(S)gene of SARS-CoV-2.We generated a luciferase(Luc)-expressing pseudovirus containing the wild-type or mutant S protein of SARS-CoV-2 in the envelope-defective HIV-1 backbone.The key parameters for this pseudovirus-based assay,including the S mutants and virus incubation time,were optimized.This pseudovirus contains a Luc reporter gene that enabled us to easily quantify virus entry into angiotensin-converting enzyme 2(ACE2)-expressing 293T cells.Cathepsin(Cat)B/L inhibitor E64d could significantly block SARS-CoV-2 pseudovirus infection in 293T-ACE2 cells.Furthermore,the SARS-CoV-2 spike pseudotyped virus could be neutralized by sera from convalescent COVID-19 patients or recombinant ACE2 with the fused Fc region of human IgG1.Thus,we developed a pseudovirus-based assay for SARS-CoV-2,which will be valuable for evaluating viral entry inhibitors and neutralizing antibodies against this highly pathogenic virus.

关 键 词：Antiviral therapeutics CORONAVIRUS Neutralizing antibodies PSEUDOVIRUS SARS-CoV-2 Spike protein 

分 类 号：R373[医药卫生—病原生物学]