tetano
Editor, Senior Moderator
Cell Host Microbe
. 2020 Jul 3;S1931-3128(20)30362-0.
doi: 10.1016/j.chom.2020.06.021. Online ahead of print.
Neutralizing Antibody and Soluble ACE2 Inhibition of a Replication-Competent VSV-SARS-CoV-2 and a Clinical Isolate of SARS-CoV-2
James Brett Case[SUP] 1 [/SUP], Paul W Rothlauf[SUP] 2 [/SUP], Rita E Chen[SUP] 3 [/SUP], Zhuoming Liu[SUP] 4 [/SUP], Haiyan Zhao[SUP] 5 [/SUP], Arthur S Kim[SUP] 3 [/SUP], Louis-Marie Bloyet[SUP] 4 [/SUP], Qiru Zeng[SUP] 4 [/SUP], Stephen Tahan[SUP] 4 [/SUP], Lindsay Droit[SUP] 4 [/SUP], Ma Xenia G Ilagan[SUP] 6 [/SUP], Michael A Tartell[SUP] 2 [/SUP], Gaya Amarasinghe[SUP] 7 [/SUP], Jeffrey P Henderson[SUP] 1 [/SUP], Shane Miersch[SUP] 8 [/SUP], Mart Ustav[SUP] 8 [/SUP], Sachdev Sidhu[SUP] 8 [/SUP], Herbert W Virgin[SUP] 9 [/SUP], David Wang[SUP] 10 [/SUP], Siyuan Ding[SUP] 4 [/SUP], Davide Corti[SUP] 11 [/SUP], Elitza S Theel[SUP] 12 [/SUP], Daved H Fremont[SUP] 13 [/SUP], Michael S Diamond[SUP] 14 [/SUP], Sean P J Whelan[SUP] 15 [/SUP]
Affiliations
Abstract
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.
Keywords: ACE2; COVID19; SARS-CoV-2; VSV; antibody; coronavirus; neutralizing; serum; surrogate assay.
. 2020 Jul 3;S1931-3128(20)30362-0.
doi: 10.1016/j.chom.2020.06.021. Online ahead of print.
Neutralizing Antibody and Soluble ACE2 Inhibition of a Replication-Competent VSV-SARS-CoV-2 and a Clinical Isolate of SARS-CoV-2
James Brett Case[SUP] 1 [/SUP], Paul W Rothlauf[SUP] 2 [/SUP], Rita E Chen[SUP] 3 [/SUP], Zhuoming Liu[SUP] 4 [/SUP], Haiyan Zhao[SUP] 5 [/SUP], Arthur S Kim[SUP] 3 [/SUP], Louis-Marie Bloyet[SUP] 4 [/SUP], Qiru Zeng[SUP] 4 [/SUP], Stephen Tahan[SUP] 4 [/SUP], Lindsay Droit[SUP] 4 [/SUP], Ma Xenia G Ilagan[SUP] 6 [/SUP], Michael A Tartell[SUP] 2 [/SUP], Gaya Amarasinghe[SUP] 7 [/SUP], Jeffrey P Henderson[SUP] 1 [/SUP], Shane Miersch[SUP] 8 [/SUP], Mart Ustav[SUP] 8 [/SUP], Sachdev Sidhu[SUP] 8 [/SUP], Herbert W Virgin[SUP] 9 [/SUP], David Wang[SUP] 10 [/SUP], Siyuan Ding[SUP] 4 [/SUP], Davide Corti[SUP] 11 [/SUP], Elitza S Theel[SUP] 12 [/SUP], Daved H Fremont[SUP] 13 [/SUP], Michael S Diamond[SUP] 14 [/SUP], Sean P J Whelan[SUP] 15 [/SUP]
Affiliations
- PMID: 32735849
- DOI: 10.1016/j.chom.2020.06.021
Abstract
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.
Keywords: ACE2; COVID19; SARS-CoV-2; VSV; antibody; coronavirus; neutralizing; serum; surrogate assay.