@article{0d2b0bb999f4488197d95472d40d29bd,
title = "Neutralizing Antibody and Soluble ACE2 Inhibition of a Replication-Competent VSV-SARS-CoV-2 and a Clinical Isolate of SARS-CoV-2",
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",
author = "Case, {James Brett} and Rothlauf, {Paul W.} and Chen, {Rita E.} and Zhuoming Liu and Haiyan Zhao and Kim, {Arthur S.} and Bloyet, {Louis Marie} and Qiru Zeng and Stephen Tahan and Lindsay Droit and Ilagan, {Ma Xenia G.} and Tartell, {Michael A.} and Gaya Amarasinghe and Henderson, {Jeffrey P.} and Shane Miersch and Mart Ustav and Sachdev Sidhu and Virgin, {Herbert W.} and David Wang and Siyuan Ding and Davide Corti and Theel, {Elitza S.} and Fremont, {Daved H.} and Diamond, {Michael S.} and Whelan, {Sean P.J.}",
note = "Funding Information: This study was supported by NIH contracts and grants (75N93019C00062, HHSN272201700060C, R01 AI127828, R37 AI059371, and U01 AI151810), the Defense Advanced Research Project Agency (HR001117S0019), and gifts from Washington University in Saint Louis. J.B.C. is supported by a Helen Hay Whitney Foundation postdoctoral fellowship. We thank Natalie Thornburg for providing the clinical isolate of SARS-CoV-2, James Rini for providing RBD used to detect phage display mAbs, Jan Carette for providing A549 and H1Hela cells, Harry Greenberg for providing Huh7.5.1 and MA104 cells, and Stanley Perlman for providing Calu-3 cells. We also thank Wandy Beatty and the Molecular Microbiology Imaging Facility at the Washington University School of Medicine for taking the electron microscopy pictures. Some of the figures were created using BioRender.com. J.B.C. performed SARS-CoV-2 neutralization experiments. P.W.R. generated and characterized VSV-SARS-CoV-2-SΔ21 and performed neutralization experiments. R.E.C. Z.L. L.M.B. M.A.T. S.D. and Q.Z. provided experimental assistance. S.T. L.D. and D.W. prepared RNA-seq libraries and assembled the VSV-SARS-CoV-2-SΔ21 sequence. H.Z. and D.H.F. generated and provided purified proteins; D.C. and H.W.V. provided the recombinant mAbs; S.M. M.U. S.S. and G.A. provided other recombinant mAbs; and E.S.T. and J.P.H. identified and provided the human immune serum. M.X.G.I. helped with automated microscope use and analysis. J.B.C. P.W.R. S.P.J.W. and M.S.D. wrote the initial draft, with the other authors providing editing comments. M.S.D. is a consultant for Inbios, Eli Lilly, Vir Biotechnology, and NGM Biopharmaceuticals and is on the Scientific Advisory Board of Moderna. The Diamond laboratory has received unrelated funding under sponsored research agreements from Moderna and Emergent BioSolutions. D.C. and H.W.V. are employees of Vir Biotechnology Inc. and may hold shares in Vir Biotechnology Inc. S.P.J.W. and P.W.R. have filed a disclosure with Washington University for the recombinant VSV. Funding Information: This study was supported by NIH contracts and grants ( 75N93019C00062 , HHSN272201700060C , R01 AI127828 , R37 AI059371 , and U01 AI151810 ), the Defense Advanced Research Project Agency ( HR001117S0019 ), and gifts from Washington University in Saint Louis. J.B.C. is supported by a Helen Hay Whitney Foundation postdoctoral fellowship. We thank Natalie Thornburg for providing the clinical isolate of SARS-CoV-2, James Rini for providing RBD used to detect phage display mAbs, Jan Carette for providing A549 and H1Hela cells, Harry Greenberg for providing Huh7.5.1 and MA104 cells, and Stanley Perlman for providing Calu-3 cells. We also thank Wandy Beatty and the Molecular Microbiology Imaging Facility at the Washington University School of Medicine for taking the electron microscopy pictures. Some of the figures were created using BioRender.com . Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",
year = "2020",
month = sep,
day = "9",
doi = "10.1016/j.chom.2020.06.021",
language = "English",
volume = "28",
pages = "475--485.e5",
journal = "Cell Host and Microbe",
issn = "1931-3128",
number = "3",
}