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
Identifieur interne : 002064 ( Main/Curation ); précédent : 002063; suivant : 002065A safe and convenient pseudovirus-based inhibition assay to detect neutralizing antibodies and screen for viral entry inhibitors against the novel human coronavirus MERS-CoV
Auteurs : Guangyu Zhao [République populaire de Chine] ; Lanying Du [États-Unis] ; Cuiqing Ma [États-Unis] ; Ye Li [États-Unis] ; Lin Li [République populaire de Chine] ; Vincent Km Poon [République populaire de Chine] ; Lili Wang [États-Unis] ; Fei Yu [États-Unis] ; Bo-Jian Zheng [République populaire de Chine] ; Shibo Jiang [États-Unis, République populaire de Chine] ; Yusen Zhou [République populaire de Chine]Source :
- Virology Journal [ 1743-422X ] ; 2013.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (isolement et purification), Anticorps neutralisants (isolement et purification), Antiviraux (isolement et purification), Coloration et marquage (), Coronavirus (), Coronavirus (immunologie), Glycoprotéine de spicule des coronavirus (génétique), Glycoprotéine de spicule des coronavirus (métabolisme), Humains, Lignée cellulaire, Luciferases (analyse), Luciferases (génétique), Souris, Tests de neutralisation (), VIH-1 (Virus de l'Immunodéficience Humaine de type 1) (génétique), Évaluation préclinique de médicament ().
- MESH :
- analyse : Luciferases.
- génétique : Glycoprotéine de spicule des coronavirus, Luciferases, VIH-1 (Virus de l'Immunodéficience Humaine de type 1).
- immunologie : Coronavirus.
- isolement et purification : Anticorps antiviraux, Anticorps neutralisants, Antiviraux.
- métabolisme : Glycoprotéine de spicule des coronavirus.
- Animaux, Coloration et marquage, Coronavirus, Humains, Lignée cellulaire, Souris, Tests de neutralisation, Évaluation préclinique de médicament.
English descriptors
- KwdEn :
- Animals, Antibodies, Neutralizing (isolation & purification), Antibodies, Viral (isolation & purification), Antiviral Agents (isolation & purification), Cell Line, Coronavirus (drug effects), Coronavirus (immunology), Drug Evaluation, Preclinical (methods), HIV-1 (genetics), Humans, Luciferases (analysis), Luciferases (genetics), Mice, Neutralization Tests (methods), Spike Glycoprotein, Coronavirus (genetics), Spike Glycoprotein, Coronavirus (metabolism), Staining and Labeling (methods).
- MESH :
- chemical , analysis : Luciferases.
- chemical , genetics : Luciferases, Spike Glycoprotein, Coronavirus.
- chemical , isolation & purification : Antibodies, Neutralizing, Antibodies, Viral, Antiviral Agents.
- drug effects : Coronavirus.
- genetics : HIV-1.
- immunology : Coronavirus.
- chemical , metabolism : Spike Glycoprotein, Coronavirus.
- methods : Drug Evaluation, Preclinical, Neutralization Tests, Staining and Labeling.
- Animals, Cell Line, Humans, Mice.
Abstract
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.
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.
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.
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.
The online version of this article (doi:10.1186/1743-422X-10-266) contains supplementary material, which is available to authorized users.
Url:
DOI: 10.1186/1743-422X-10-266
PubMed: 23978242
PubMed Central: 3765664
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PMC:3765664Le document en format XML
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<author><name sortKey="Wang, Lili" sort="Wang, Lili" uniqKey="Wang L" first="Lili" last="Wang">Lili Wang</name>
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<author><name sortKey="Yu, Fei" sort="Yu, Fei" uniqKey="Yu F" first="Fei" last="Yu">Fei Yu</name>
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<author><name sortKey="Zheng, Bo Jian" sort="Zheng, Bo Jian" uniqKey="Zheng B" first="Bo-Jian" last="Zheng">Bo-Jian Zheng</name>
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<author><name sortKey="Zhou, Yusen" sort="Zhou, Yusen" uniqKey="Zhou Y" first="Yusen" last="Zhou">Yusen Zhou</name>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Antibodies, Neutralizing (isolation & purification)</term>
<term>Antibodies, Viral (isolation & purification)</term>
<term>Antiviral Agents (isolation & purification)</term>
<term>Cell Line</term>
<term>Coronavirus (drug effects)</term>
<term>Coronavirus (immunology)</term>
<term>Drug Evaluation, Preclinical (methods)</term>
<term>HIV-1 (genetics)</term>
<term>Humans</term>
<term>Luciferases (analysis)</term>
<term>Luciferases (genetics)</term>
<term>Mice</term>
<term>Neutralization Tests (methods)</term>
<term>Spike Glycoprotein, Coronavirus (genetics)</term>
<term>Spike Glycoprotein, Coronavirus (metabolism)</term>
<term>Staining and Labeling (methods)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term>
<term>Anticorps antiviraux (isolement et purification)</term>
<term>Anticorps neutralisants (isolement et purification)</term>
<term>Antiviraux (isolement et purification)</term>
<term>Coloration et marquage ()</term>
<term>Coronavirus ()</term>
<term>Coronavirus (immunologie)</term>
<term>Glycoprotéine de spicule des coronavirus (génétique)</term>
<term>Glycoprotéine de spicule des coronavirus (métabolisme)</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Luciferases (analyse)</term>
<term>Luciferases (génétique)</term>
<term>Souris</term>
<term>Tests de neutralisation ()</term>
<term>VIH-1 (Virus de l'Immunodéficience Humaine de type 1) (génétique)</term>
<term>Évaluation préclinique de médicament ()</term>
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</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Luciferases</term>
<term>Spike Glycoprotein, Coronavirus</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Antibodies, Neutralizing</term>
<term>Antibodies, Viral</term>
<term>Antiviral Agents</term>
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<term>Luciferases</term>
<term>VIH-1 (Virus de l'Immunodéficience Humaine de type 1)</term>
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<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Coronavirus</term>
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<keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Coronavirus</term>
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<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Anticorps antiviraux</term>
<term>Anticorps neutralisants</term>
<term>Antiviraux</term>
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<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Spike Glycoprotein, Coronavirus</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Drug Evaluation, Preclinical</term>
<term>Neutralization Tests</term>
<term>Staining and Labeling</term>
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<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glycoprotéine de spicule des coronavirus</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Cell Line</term>
<term>Humans</term>
<term>Mice</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Coloration et marquage</term>
<term>Coronavirus</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Souris</term>
<term>Tests de neutralisation</term>
<term>Évaluation préclinique de médicament</term>
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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p>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.</p>
</sec>
<sec><title>Methods</title>
<p>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.</p>
</sec>
<sec><title>Results</title>
<p>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.</p>
</sec>
<sec><title>Conclusion</title>
<p>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.</p>
</sec>
<sec><title>Electronic supplementary material</title>
<p>The online version of this article (doi:10.1186/1743-422X-10-266) contains supplementary material, which is available to authorized users.</p>
</sec>
</div>
</front>
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