Human Immunodeficiency Viruses Pseudotyped with SARS-CoV-2 Spike Proteins Infect a Broad Spectrum of Human Cell Lines through Multiple Entry Mechanisms.
Human Immunodeficiency Viruses Pseudotyped with SARS-CoV-2 Spike Proteins Infect a Broad Spectrum of Human Cell Lines through Multiple Entry Mechanisms.
复制标题
带有SARS-CoV-2刺突蛋白假型的人类免疫缺陷病毒通过多种进入机制感染广泛的人类细胞系
DOI：
10.3390/v13060953
复制
发表时间：
2021-05-21
期刊：
影响因子：
--
通讯作者：
Chang TL
中科院分区：
文献类型：
--
作者：
Xu C;Wang A;Geng K;Honnen W;Wang X;Bruiners N;Singh S;Ferrara F;D'Angelo S;Bradbury ARM;Gennaro ML;Liu D;Pinter A;Chang TL
严重急性呼吸综合征相关冠状病毒（SARS-CoV-2）是冠状病毒病19（COVID-19）的病原体，通过表面/刺突（S）蛋白中的受体结合结构域（RBD）附着于人血管紧张素转换酶2（hACE 2）进入细胞。有几种表达SARS-CoV-2 S蛋白的假型病毒可用，但其中许多只能感染hACE 2过表达的细胞系。在这里，我们报告了使用一个简单的，两个质粒，假型病毒系统，包括一个SARS-CoV-2刺突表达质粒和HIV载体与或不与vpr调查的SARS-CoV-2进入事件在各种细胞系。当使用不含vpr的HIV载体时，在胎牛血清（FBS）存在下产生的假型SARS-CoV-2病毒仅能够感染工程化hACE 2过表达细胞系，而在无血清条件下产生的病毒能够感染更广泛的细胞，包括没有hACE 2过表达的细胞。当使用含有vpr的HIV载体时，假型病毒能够感染广谱的细胞类型，无论病毒是否在存在或不存在FBS的情况下产生。各种细胞类型的感染敏感性与hACE 2、TMPRSS 2或TMPRSS 4的mRNA丰度无关。假型SARS-CoV-2病毒和有复制能力的SARS-CoV-2病毒对具有高丰度hACE 2的细胞中的抗刺突RBD抗体的中和同样敏感。然而，抗刺突RBD抗体不阻断假型病毒进入具有低丰度hACE 2的细胞系。我们进一步发现，CD 147参与了病毒进入具有低丰度hACE 2的A549细胞。因此，我们的检测方法可用于药物和抗体筛选，以及用于研究细胞受体，包括hACE 2，CD 147和酪氨酸蛋白激酶受体UFO（AXL），用于SARS-CoV-2进入各种细胞系的事件。
Severe acute respiratory syndrome-related coronavirus (SARS-CoV-2), the causative agent of coronavirus disease 19 (COVID-19), enters cells through attachment to the human angiotensin converting enzyme 2 (hACE2) via the receptor-binding domain (RBD) in the surface/spike (S) protein. Several pseudotyped viruses expressing SARS-CoV-2 S proteins are available, but many of these can only infect hACE2-overexpressing cell lines. Here, we report the use of a simple, two-plasmid, pseudotyped virus system comprising a SARS-CoV-2 spike-expressing plasmid and an HIV vector with or without vpr to investigate the SARS-CoV-2 entry event in various cell lines. When an HIV vector without vpr was used, pseudotyped SARS-CoV-2 viruses produced in the presence of fetal bovine serum (FBS) were able to infect only engineered hACE2-overexpressing cell lines, whereas viruses produced under serum-free conditions were able to infect a broader range of cells, including cells without hACE2 overexpression. When an HIV vector containing vpr was used, pseudotyped viruses were able to infect a broad spectrum of cell types regardless of whether viruses were produced in the presence or absence of FBS. Infection sensitivities of various cell types did not correlate with mRNA abundance of hACE2, TMPRSS2, or TMPRSS4. Pseudotyped SARS-CoV-2 viruses and replication-competent SARS-CoV-2 virus were equally sensitive to neutralization by an anti-spike RBD antibody in cells with high abundance of hACE2. However, the anti-spike RBD antibody did not block pseudotyped viral entry into cell lines with low abundance of hACE2. We further found that CD147 was involved in viral entry in A549 cells with low abundance of hACE2. Thus, our assay is useful for drug and antibody screening as well as for investigating cellular receptors, including hACE2, CD147, and tyrosine-protein kinase receptor UFO (AXL), for the SARS-CoV-2 entry event in various cell lines.
登录
查看更多内容
DOI：
10.1126/science.abd3072
发表时间：
2020-11-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Daly JL;Simonetti B;Klein K;Chen KE;Williamson MK;Antón-Plágaro C;Shoemark DK;Simón-Gracia L;Bauer M;Hollandi R;Greber UF;Horvath P;Sessions RB;Helenius A;Hiscox JA;Teesalu T;Matthews DA;Davidson AD;Collins BM;Cullen PJ;Yamauchi Y
通讯作者：
Yamauchi Y
影响因子：
64.5
作者：
Clausen, Thomas Mandel;Sandoval, Daniel R.;Esko, Jeffrey D.
通讯作者：
Esko, Jeffrey D.
影响因子：
3.3
作者：
Rapista A;Ding J;Benito B;Lo YT;Neiditch MB;Lu W;Chang TL
通讯作者：
Chang TL
BNT162B2 mRNA COVID-19疫苗的安全性和功效。
DOI：
10.1056/nejmoa2034577
发表时间：
2020-12-31
期刊：
The New England journal of medicine
影响因子：
--
作者：
Polack FP;Thomas SJ;Kitchin N;Absalon J;Gurtman A;Lockhart S;Perez JL;Pérez Marc G;Moreira ED;Zerbini C;Bailey R;Swanson KA;Roychoudhury S;Koury K;Li P;Kalina WV;Cooper D;Frenck RW Jr;Hammitt LL;Türeci Ö;Nell H;Schaefer A;Ünal S;Tresnan DB;Mather S;Dormitzer PR;Şahin U;Jansen KU;Gruber WC;C4591001 Clinical Trial Group
通讯作者：
C4591001 Clinical Trial Group
影响因子：
3.3
作者：
Kogan M;Rappaport J
通讯作者：
Rappaport J