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

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• 作者：Guangyu Zhao (5)
  Lanying Du (5)
  Cuiqing Ma (5)
  Ye Li (5)
  Lin Li (5)
  Vincent KM Poon (5)
  Lili Wang (5)
  Fei Yu (5)
  Bo-Jian Zheng (5)
  Shibo Jiang (5) (5)
  Yusen Zhou (5)
  
• 关键词：Novel human coronavirus ; MERS ; CoV ; Spike protein ; Pseudovirus ; Neutralizing antibodies ; Antiviral therapeutics
• 刊名：Virology Journal
• 出版年：2013
• 出版时间：December 2013
• 年：2013
• 卷：10
• 期：1
• 全文大小：306KB
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• 作者单位：Guangyu Zhao (5)
  Lanying Du (5)
  Cuiqing Ma (5)
  Ye Li (5)
  Lin Li (5)
  Vincent KM Poon (5)
  Lili Wang (5)
  Fei Yu (5)
  Bo-Jian Zheng (5)
  Shibo Jiang (5) (5)
  Yusen Zhou (5)
  
  5. Key Laboratory of Medical Molecular Virology of Ministries of Education and Health, Shanghai Medical College and Institute of Medical Microbiology, Fudan University, Shanghai, China
  
• ISSN：1743-422X

文摘

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 377-62) 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.