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Published in: Virology Journal 1/2021

Open Access 01-12-2021 | SARS-CoV-2 | Research

Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein

Authors: Hideki Tani, Miyuki Kimura, Long Tan, Yoshihiro Yoshida, Tatsuhiko Ozawa, Hiroyuki Kishi, Shuetsu Fukushi, Masayuki Saijo, Kaori Sano, Tadaki Suzuki, Hitoshi Kawasuji, Akitoshi Ueno, Yuki Miyajima, Yasutaka Fukui, Ippei Sakamaki, Yoshihiro Yamamoto, Yoshitomo Morinaga

Published in: Virology Journal | Issue 1/2021

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Abstract

Background

SARS-CoV-2 is a novel coronavirus that emerged in 2019 and is now classified in the genus Coronavirus with closely related SARS-CoV. SARS-CoV-2 is highly pathogenic in humans and is classified as a biosafety level (BSL)-3 pathogen, which makes manipulating it relatively difficult due to its infectious nature.

Methods

To circumvent the need for BSL-3 laboratories, an alternative assay was developed that avoids live virus and instead uses a recombinant VSV expressing luciferase and possesses the full length or truncated spike proteins of SARS-CoV-2. Furthermore, to measure SARS-CoV-2 neutralizing antibodies under BSL2 conditions, a chemiluminescence reduction neutralization test (CRNT) for SARS-CoV-2 was developed. The neutralization values of the serum samples collected from hospitalized patients with COVID-19 or SARS-CoV-2 PCR-negative donors against the pseudotyped virus infection evaluated by the CRNT were compared with antibody titers determined from an enzyme-linked immunosorbent assay (ELISA) or an immunofluorescence assay (IFA).

Results

The CRNT, which used whole blood collected from hospitalized patients with COVID-19, was also examined. As a result, the inhibition of pseudotyped virus infection was specifically observed in both serum and whole blood and was also correlated with the results of the IFA.

Conclusions

In conclusion, the CRNT for COVID-19 is a convenient assay system that can be performed in a BSL-2 laboratory with high specificity and sensitivity for evaluating the occurrence of neutralizing antibodies against SARS-CoV-2.
Literature
1.
go back to reference Huang C, Wang Y, Li X, Ren L, Zhao J, Hu Y, Zhang L, Fan G, Xu J, Gu X, et al. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet. 2020;395:497–506.CrossRef Huang C, Wang Y, Li X, Ren L, Zhao J, Hu Y, Zhang L, Fan G, Xu J, Gu X, et al. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet. 2020;395:497–506.CrossRef
2.
go back to reference Musa A, Pendi K, Hashemi A, Warbasse E, Kouyoumjian S, Yousif J, Blodget E, Stevens S, Aly B, Baron DA. Remdesivir for the treatment of COVID-19: a systematic review of the literature. West J Emerg Med. 2020;21:737–41.PubMedPubMedCentral Musa A, Pendi K, Hashemi A, Warbasse E, Kouyoumjian S, Yousif J, Blodget E, Stevens S, Aly B, Baron DA. Remdesivir for the treatment of COVID-19: a systematic review of the literature. West J Emerg Med. 2020;21:737–41.PubMedPubMedCentral
3.
go back to reference Tu H, Tu S, Gao S, Shao A, Sheng J. Current epidemiological and clinical features of COVID-19; a global perspective from China. J Infect. 2020;81:1–9.CrossRef Tu H, Tu S, Gao S, Shao A, Sheng J. Current epidemiological and clinical features of COVID-19; a global perspective from China. J Infect. 2020;81:1–9.CrossRef
4.
go back to reference Jin Y, Yang H, Ji W, Wu W, Chen S, Zhang W, Duan G. Virology, epidemiology, pathogenesis, and control of COVID-19. Viruses. 2020;12:372.CrossRef Jin Y, Yang H, Ji W, Wu W, Chen S, Zhang W, Duan G. Virology, epidemiology, pathogenesis, and control of COVID-19. Viruses. 2020;12:372.CrossRef
5.
go back to reference Li Q, Liu Q, Huang W, Li X, Wang Y. Current status on the development of pseudoviruses for enveloped viruses. Rev Med Virol. 2018;28:e1963.CrossRef Li Q, Liu Q, Huang W, Li X, Wang Y. Current status on the development of pseudoviruses for enveloped viruses. Rev Med Virol. 2018;28:e1963.CrossRef
6.
go back to reference Joglekar AV, Sandoval S. Pseudotyped lentiviral vectors: one vector. Many Guises Hum Gene Ther Methods. 2017;28:291–301.CrossRef Joglekar AV, Sandoval S. Pseudotyped lentiviral vectors: one vector. Many Guises Hum Gene Ther Methods. 2017;28:291–301.CrossRef
7.
go back to reference Whitt MA. Generation of VSV pseudotypes using recombinant deltaG-VSV for studies on virus entry, identification of entry inhibitors, and immune responses to vaccines. J Virol Methods. 2010;169:365–74.CrossRef Whitt MA. Generation of VSV pseudotypes using recombinant deltaG-VSV for studies on virus entry, identification of entry inhibitors, and immune responses to vaccines. J Virol Methods. 2010;169:365–74.CrossRef
8.
go back to reference Nie J, Li Q, Wu J, Zhao C, Hao H, Liu H, Zhang L, Nie L, Qin H, Wang M, et al. Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2. Emerg Microbes Infect. 2020;9:680–6.CrossRef Nie J, Li Q, Wu J, Zhao C, Hao H, Liu H, Zhang L, Nie L, Qin H, Wang M, et al. Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2. Emerg Microbes Infect. 2020;9:680–6.CrossRef
9.
go back to reference Case JB, Rothlauf PW, Chen RE, Liu Z, Zhao H, Kim AS, Bloyet LM, Zeng Q, Tahan S, Droit L, et al. Neutralizing antibody and soluble ACE2 inhibition of a replication-competent VSV-SARS-CoV-2 and a clinical isolate of SARS-CoV-2. Cell Host Microbe. 2020;28(475–485):e475.CrossRef Case JB, Rothlauf PW, Chen RE, Liu Z, Zhao H, Kim AS, Bloyet LM, Zeng Q, Tahan S, Droit L, et al. Neutralizing antibody and soluble ACE2 inhibition of a replication-competent VSV-SARS-CoV-2 and a clinical isolate of SARS-CoV-2. Cell Host Microbe. 2020;28(475–485):e475.CrossRef
10.
go back to reference Zettl F, Meister TL, Vollmer T, Fischer B, Steinmann J, Krawczyk A, V’Kovski P, Todt D, Steinmann E, Pfaender S, Zimmer G. Rapid quantification of SARS-CoV-2-neutralizing antibodies using propagation-defective vesicular stomatitis virus pseudotypes. Vaccines (Basel). 2020;8:386.CrossRef Zettl F, Meister TL, Vollmer T, Fischer B, Steinmann J, Krawczyk A, V’Kovski P, Todt D, Steinmann E, Pfaender S, Zimmer G. Rapid quantification of SARS-CoV-2-neutralizing antibodies using propagation-defective vesicular stomatitis virus pseudotypes. Vaccines (Basel). 2020;8:386.CrossRef
11.
go back to reference Tan CW, Chia WN, Qin X, Liu P, Chen MI, Tiu C, Hu Z, Chen VC, Young BE, Sia WR, et al. A SARS-CoV-2 surrogate virus neutralization test based on antibody-mediated blockage of ACE2-spike protein-protein interaction. Nat Biotechnol. 2020;38:1073–8.CrossRef Tan CW, Chia WN, Qin X, Liu P, Chen MI, Tiu C, Hu Z, Chen VC, Young BE, Sia WR, et al. A SARS-CoV-2 surrogate virus neutralization test based on antibody-mediated blockage of ACE2-spike protein-protein interaction. Nat Biotechnol. 2020;38:1073–8.CrossRef
12.
go back to reference Almahboub SA, Algaissi A, Alfaleh MA, ElAssouli MZ, Hashem AM. Evaluation of neutralizing antibodies against highly pathogenic coronaviruses: a detailed protocol for a rapid evaluation of neutralizing antibodies using vesicular stomatitis virus pseudovirus-based assay. Front Microbiol. 2020;2020:11. Almahboub SA, Algaissi A, Alfaleh MA, ElAssouli MZ, Hashem AM. Evaluation of neutralizing antibodies against highly pathogenic coronaviruses: a detailed protocol for a rapid evaluation of neutralizing antibodies using vesicular stomatitis virus pseudovirus-based assay. Front Microbiol. 2020;2020:11.
13.
go back to reference Saijo M, Qing T, Niikura M, Maeda A, Ikegami T, Prehaud C, Kurane I, Morikawa S. Recombinant nucleoprotein-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus. J Clin Microbiol. 2002;40:1587–91.CrossRef Saijo M, Qing T, Niikura M, Maeda A, Ikegami T, Prehaud C, Kurane I, Morikawa S. Recombinant nucleoprotein-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus. J Clin Microbiol. 2002;40:1587–91.CrossRef
14.
go back to reference Fukushi S, Mizutani T, Saijo M, Matsuyama S, Miyajima N, Taguchi F, Itamura S, Kurane I, Morikawa S. Vesicular stomatitis virus pseudotyped with severe acute respiratory syndrome coronavirus spike protein. J Gen Virol. 2005;86:2269–74.CrossRef Fukushi S, Mizutani T, Saijo M, Matsuyama S, Miyajima N, Taguchi F, Itamura S, Kurane I, Morikawa S. Vesicular stomatitis virus pseudotyped with severe acute respiratory syndrome coronavirus spike protein. J Gen Virol. 2005;86:2269–74.CrossRef
15.
go back to reference Kawase M, Shirato K, Matsuyama S, Taguchi F. Protease-mediated entry via the endosome of human coronavirus 229E. J Virol. 2009;83:712–21.CrossRef Kawase M, Shirato K, Matsuyama S, Taguchi F. Protease-mediated entry via the endosome of human coronavirus 229E. J Virol. 2009;83:712–21.CrossRef
16.
go back to reference Tani H, Shiokawa M, Kaname Y, Kambara H, Mori Y, Abe T, Moriishi K, Matsuura Y. Involvement of ceramide in the propagation of Japanese encephalitis virus. J Virol. 2010;84:2798–807.CrossRef Tani H, Shiokawa M, Kaname Y, Kambara H, Mori Y, Abe T, Moriishi K, Matsuura Y. Involvement of ceramide in the propagation of Japanese encephalitis virus. J Virol. 2010;84:2798–807.CrossRef
17.
go back to reference Shirato K, Nao N, Katano H, Takayama I, Saito S, Kato F, Katoh H, Sakata M, Nakatsu Y, Mori Y, et al: Development of genetic diagnostic methods for detection for novel coronavirus 2019(nCoV-2019) in Japan. Jpn J Infect Dis 2020; 73. Shirato K, Nao N, Katano H, Takayama I, Saito S, Kato F, Katoh H, Sakata M, Nakatsu Y, Mori Y, et al: Development of genetic diagnostic methods for detection for novel coronavirus 2019(nCoV-2019) in Japan. Jpn J Infect Dis 2020; 73.
18.
go back to reference Amanat F, Stadlbauer D, Strohmeier S, Nguyen THO, Chromikova V, McMahon M, Jiang K, Arunkumar GA, Jurczyszak D, Polanco J, et al. A serological assay to detect SARS-CoV-2 seroconversion in humans. Nat Med. 2020;26:1033–6.CrossRef Amanat F, Stadlbauer D, Strohmeier S, Nguyen THO, Chromikova V, McMahon M, Jiang K, Arunkumar GA, Jurczyszak D, Polanco J, et al. A serological assay to detect SARS-CoV-2 seroconversion in humans. Nat Med. 2020;26:1033–6.CrossRef
19.
go back to reference Sakata M, Tani H, Anraku M, Kataoka M, Nagata N, Seki F, Tahara M, Otsuki N, Okamoto K, Takeda M, Mori Y. Analysis of VSV pseudotype virus infection mediated by rubella virus envelope proteins. Sci Rep. 2017;7:11607.CrossRef Sakata M, Tani H, Anraku M, Kataoka M, Nagata N, Seki F, Tahara M, Otsuki N, Okamoto K, Takeda M, Mori Y. Analysis of VSV pseudotype virus infection mediated by rubella virus envelope proteins. Sci Rep. 2017;7:11607.CrossRef
20.
go back to reference Tani H, Iha K, Shimojima M, Fukushi S, Taniguchi S, Yoshikawa T, Kawaoka Y, Nakasone N, Ninomiya H, Saijo M, Morikawa S. Analysis of Lujo virus cell entry using pseudotype vesicular stomatitis virus. J Virol. 2014;88:7317–30.CrossRef Tani H, Iha K, Shimojima M, Fukushi S, Taniguchi S, Yoshikawa T, Kawaoka Y, Nakasone N, Ninomiya H, Saijo M, Morikawa S. Analysis of Lujo virus cell entry using pseudotype vesicular stomatitis virus. J Virol. 2014;88:7317–30.CrossRef
21.
go back to reference Tani H, Shimojima M, Fukushi S, Yoshikawa T, Fukuma A, Taniguchi S, Morikawa S, Saijo M. Characterization of glycoprotein-mediated entry of severe fever with thrombocytopenia syndrome virus. J Virol. 2016;90:5292–301.CrossRef Tani H, Shimojima M, Fukushi S, Yoshikawa T, Fukuma A, Taniguchi S, Morikawa S, Saijo M. Characterization of glycoprotein-mediated entry of severe fever with thrombocytopenia syndrome virus. J Virol. 2016;90:5292–301.CrossRef
22.
go back to reference Crawford KHD, Eguia R, Dingens AS, Loes AN, Malone KD, Wolf CR, Chu HY, Tortorici MA, Veesler D, Murphy M, et al. Protocol and reagents for pseudotyping lentiviral particles with SARS-CoV-2 spike protein for neutralization assays. Viruses. 2020;12:513.CrossRef Crawford KHD, Eguia R, Dingens AS, Loes AN, Malone KD, Wolf CR, Chu HY, Tortorici MA, Veesler D, Murphy M, et al. Protocol and reagents for pseudotyping lentiviral particles with SARS-CoV-2 spike protein for neutralization assays. Viruses. 2020;12:513.CrossRef
Metadata
Title
Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein
Authors
Hideki Tani
Miyuki Kimura
Long Tan
Yoshihiro Yoshida
Tatsuhiko Ozawa
Hiroyuki Kishi
Shuetsu Fukushi
Masayuki Saijo
Kaori Sano
Tadaki Suzuki
Hitoshi Kawasuji
Akitoshi Ueno
Yuki Miyajima
Yasutaka Fukui
Ippei Sakamaki
Yoshihiro Yamamoto
Yoshitomo Morinaga
Publication date
01-12-2021
Publisher
BioMed Central
Published in
Virology Journal / Issue 1/2021
Electronic ISSN: 1743-422X
DOI
https://doi.org/10.1186/s12985-021-01490-7

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