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Open Access 01-12-2013 | Methodology

A safe and convenient pseudovirus-based inhibition assay to detect neutralizing antibodies and screen for viral entry inhibitors against the novel human coronavirus MERS-CoV

Authors: Guangyu Zhao, Lanying Du, Cuiqing Ma, Ye Li, Lin Li, Vincent KM Poon, Lili Wang, Fei Yu, Bo-Jian Zheng, Shibo Jiang, Yusen Zhou

Published in: Virology Journal | Issue 1/2013

Abstract

Background

Evidence points to the emergence of a novel human coronavirus, Middle East respiratory syndrome coronavirus (MERS-CoV), which causes a severe acute respiratory syndrome (SARS)-like disease. In response, the development of effective vaccines and therapeutics remains a clinical priority. To accomplish this, it is necessary to evaluate neutralizing antibodies and screen for MERS-CoV entry inhibitors.

Methods

In this study, we produced a pseudovirus bearing the full-length spike (S) protein of MERS-CoV in the Env-defective, luciferase-expressing HIV-1 backbone. We then established a pseudovirus-based inhibition assay to detect neutralizing antibodies and anti-MERS-CoV entry inhibitors.

Results

Our results demonstrated that the generated MERS-CoV pseudovirus allows for single-cycle infection of a variety of cells expressing dipeptidyl peptidase-4 (DPP4), the confirmed receptor for MERS-CoV. Consistent with the results from a live MERS-CoV-based inhibition assay, the antisera of mice vaccinated with a recombinant protein containing receptor-binding domain (RBD, residues 377–662) of MERS-CoV S fused with Fc of human IgG exhibited neutralizing antibody response against infection of MERS-CoV pseudovirus. Furthermore, one small molecule HIV entry inhibitor targeting gp41 (ADS-J1) and the 3-hydroxyphthalic anhydride-modified human serum albumin (HP-HSA) could significantly inhibit MERS-CoV pseudovirus infection.

Conclusion

Taken together, the established MERS-CoV inhibition assay is a safe and convenient pseudovirus-based alternative to BSL-3 live-virus restrictions and can be used to rapidly screen MERS-CoV entry inhibitors, as well as evaluate vaccine-induced neutralizing antibodies against the highly pathogenic MERS-CoV.

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Title: A safe and convenient pseudovirus-based inhibition assay to detect neutralizing antibodies and screen for viral entry inhibitors against the novel human coronavirus MERS-CoV
Authors: Guangyu Zhao
Lanying Du
Cuiqing Ma
Ye Li
Lin Li
Vincent KM Poon
Lili Wang
Fei Yu
Bo-Jian Zheng
Shibo Jiang
Yusen Zhou
Publication date: 01-12-2013
Publisher: BioMed Central
Published in: Virology Journal / Issue 1/2013
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/1743-422X-10-266

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