Neutralizing Antibody and Soluble ACE2 Inhibition of a Replication-Competent VSV-SARS-CoV-2 and a Clinical Isolate of SARS-CoV-2
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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.
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SARS-CoV-2 and SARS-CoV enter host cells through binding of the S glycoprotein to angiotensin converting enzyme 2 (ACE2).
“SARS-CoV-2 and SARS-CoV enter host cells through binding of the S glycoprotein to angiotensin converting enzyme 2 (ACE2)1,28.”
From Sensitivity of SARS-CoV-2 B.1.1.7 to mRNA vaccine-elicited antibodies (2021) · cites this paper as [28]
The pseudovirus-neutralization test (pVNT) has been shown to be in close concordance with live SARS-CoV-2-neutralization assays.
“a pseudovirus-neutralization test (pVNT) that has been shown to be in close concordance with live SARS-CoV-2–neutralization assays (17, 18)”
From Neutralization of SARS-CoV-2 Omicron by BNT162b2 mRNA vaccine–elicited human sera (2022) · cites this paper as [17]
The study used two orthogonal test systems: a pseudovirus-neutralization test and a live SARS-CoV-2-neutralization test.
“we used two orthogonal test systems: a pseudovirus-neutralization test (pVNT) that has been shown to be in close concordance with live SARS-CoV-2–neutralization assays (17, 18), as well as a live SARS-CoV-2–neutralization test (VNT).”
From Neutralization of SARS-CoV-2 Omicron by BNT162b2 mRNA vaccine–elicited human sera (2022) · cites this paper as [17]
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Heat shock protein 70 (Hsp70) is involved in the Zika virus cellular infection processPreprint
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Synthetic antibodies neutralize SARS-CoV-2 infection of mammalian cellsPreprint
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Self-citation
Flagged first, then by citations
Cited by
Sorted by citations
Greaney, A. J., Starr, T. N., Gilchuk, P., Zost, S. J., Binshtein, E., Loes, A. N., Hilton, S. K., Huddleston, J., Eguia, R., Crawford, K. H. D., et al. (2021) · Cell Host & Microbe · doi:10.1016/j.chom.2020.11.007 · Cited by 1,016 · see citing works
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“Selection of SARS-CoV-2 monoclonal antibody escape mutants (MARMS) VSV-SARS-CoV-2 chimera was used to select for SARS-CoV-2 S monoclonal antibody resistant mutants (MARMS) as previously described (Case et al., 2020; Liu et al., 2020b).”
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Turner, J. S., O’Halloran, J. A., Kalaidina, E., Kim, W., Schmitz, A. J., Zhou, J. Q., Lei, T., Thapa, M., Chen, R. E., Case, J. B., et al. (2021) · Nature · doi:10.1038/s41586-021-03738-2 · Cited by 831 · see citing works
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Collier, D. A., De Marco, A., Ferreira, I. A. T. M., Meng, B., Datir, R. P., Walls, A. C., Kemp, S. A., Bassi, J., Pinto, D., Silacci-Fregni, C., et al. (2021) · Nature · doi:10.1038/s41586-021-03412-7 · Cited by 728 · see citing works
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Ultrapotent human antibodies protect against SARS-CoV-2 challenge via multiple mechanisms
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Morphological, cellular and molecular basis of brain infection in COVID-19 patients
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Measuring immunity to SARS-CoV-2 infection: comparing assays and animal models
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Schmitz, A. J., Turner, J. S., Liu, Z., Aziati, I. D., Chen, R. E., Joshi, A., Bricker, T. L., Darling, T. L., Adelsberg, D. C., Alsoussi, W. B., et al. (2021) · doi:10.1101/2021.03.24.436864 · Cited by 72 · see citing works
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Self-citation
- Authors
- Case, J. B., Rothlauf, P. W., Chen, R. E., Liu, Z., Zhao, H., Kim, A. S., Bloyet, L.-M., Zeng, Q., Tahan, S., Droit, L., Ilagan, M. X. G., Tartell, M. A., Amarasinghe, G., Henderson, J. P., Miersch, S., Ustav, M., Sidhu, S., Virgin, H. W., Wang, D., Ding, S., Corti, D., Theel, E. S., Fremont, D. H., Diamond, M. S., & Whelan, S. P. J.
- Affiliations
- Washington University in St. Louis (US)
- Harvard University (US)
- University of Toronto (CA)
- VIR Biotechnology (United States) (US)
- Vir Biotechnology (Switzerland) (CH)
- Mayo Clinic (US)
- Corresponding author
- Diamond, Michael S.
- Whelan, Sean P.J.
- ORCID
- Case, James Brett
- Rothlauf, Paul W.
- Chen, Rita E.
- Liu, Zhuoming
- Zhao, Haiyan
- Kim, Arthur S.
- Bloyet, Louis-Marie
- Zeng, Qiru
- Tahan, Stephen
- Droit, Lindsay
- Ilagan, Ma. Xenia G.
- Tartell, Michael A.
- Amarasinghe, Gaya
- Henderson, Jeffrey P.
- Miersch, Shane
- Ustav, Mart
- Sidhu, Sachdev
- Virgin, Herbert W.
- Wang, David
- Ding, Siyuan
- Corti, Davide
- Theel, Elitza S.
- Fremont, Daved H.
- Diamond, Michael S.
- Whelan, Sean P.J.
- Venue
- Cell Host & Microbe
- Publisher
- Elsevier BV
- Published
- 2020-09
- Volume
- 28
- Issue
- 3
- Pages
- 475-485.e5
- ISSN
- 1931-3128 (ISSN-L)
- PMC
- PMC7332453
- OpenAlex
- W3040017923
- Cited by
- 410
- References
- 92
- Citations by year
- 2026: 25 · 2025: 25 · 2024: 40 · 2023: 47 · 2022: 99 · 2021: 127 · 2020: 46
- FWCI
- 10.46
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell Host & Microbe (publishedVersion)
- SSRN Electronic Journal (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (submittedVersion)
- HAL (submittedVersion)
- Funders
- Helen Hay Whitney Foundation (MA104)
- Washington University in St. Louis
- Washington University School of Medicine in St. Louis
- National Institutes of Health (75N93019C00062, HHSN272201700060C, R01 AI127828, R37 AI059371, U01 AI151810)
- Vir Biotechnology
- Eli Lilly and Company
- Defense Advanced Research Projects Agency (HR001117S0019)
- Topics
- Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 and COVID-19 Research
- Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Virus Infections Studies
- Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 detection and testing
- Keywords
- SARS-CoV-2, neutralizing antibodies, soluble ACE2, spike protein, focus reduction neutralization test, correlates of protection, vaccine development, viral entry inhibitors, biosafety level 2, biosafety level 3
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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
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