Neutralizing Antibody and Soluble ACE2 Inhibition of a Replication-Competent VSV-SARS-CoV-2 and a Clinical Isolate of SARS-CoV-2
Neutralizing Antibody and Soluble ACE2 Inhibition of a Replication-Competent VSV-SARS-CoV-2 and a Clinical Isolate of SARS-CoV-2
复制标题
DOI：
10.1016/j.chom.2020.06.021
复制
发表时间：
2020-09-09
影响因子：
30.3
通讯作者：
Whelan, Sean P. J.
中科院分区：
文献类型：
--
作者：
Case, James Brett;Rothlauf, Paul W.;Whelan, Sean P. J.
基于抗体的SARS-CoV-2干预措施可以限制发病率、死亡率和可能的传播。这种对抗措施的预期相关性是针对SARS-CoV-2刺突蛋白的中和抗体的水平，该刺突蛋白与宿主ACE 2受体结合以进入。使用表达eGFP的水泡性口炎病毒（VSV）的感染性分子克隆作为感染标志物，我们用SARS-CoV-2的刺突蛋白（VSV-eGFP-SARS-CoV-2）替换糖蛋白基因（G），并开发了生物安全级别为2的基于高通量成像的中和试验。我们还开发了一个焦点减少中和试验与临床分离的SARS-CoV-2在生物安全水平3。在两种测定中比较各种抗体和ACE 2-Fc可溶性诱饵蛋白的中和活性显示高度一致性。这些检测将有助于确定基于抗体的对策和疫苗对SARS-CoV-2的保护相关性。此外，可复制的VSV-eGFP-SARS-CoV-2提供了一种工具，用于在降低的生物安全控制下测试SARS-CoV-2介导的进入抑制剂。
Antibody-based interventions against SARS-CoV-2 could limit morbidity, mortality, and possibly transmission. An anticipated correlate of such countermeasures is the level of neutralizing antibodies against the SARS-CoV-2 spike protein, which engages with host ACE2 receptor for entry. Using an infectious molecular clone of vesicular stomatitis virus (VSV) expressing eGFP as a marker of infection, we replaced the glycoprotein gene (G) with the spike protein of SARS-CoV-2 (VSV-eGFP-SARS-CoV-2) and developed a high-throughput-imaging-based neutralization assay at biosafety level 2. We also developed a focus-reduction neutralization test with a clinical isolate of SARS-CoV-2 at biosafety level 3. Comparing the neutralizing activities of various antibodies and ACE2-Fc soluble decoy protein in both assays revealed a high degree of concordance. These assays will help define correlates of protection for antibody-based countermeasures and vaccines against SARS-CoV-2. Additionally, replication-competent VSV-eGFP-SARS-CoV-2 provides a tool for testing inhibitors of SARS-CoV-2 mediated entry under reduced biosafety containment.