Identifying epitopes responsible for neutralizing antibody and DC-SIGN binding on the spike glycoprotein of the severe acute respiratory syndrome coronavirus.
Identifieur interne : 001733 ( Ncbi/Checkpoint ); précédent : 001732; suivant : 001734Identifying epitopes responsible for neutralizing antibody and DC-SIGN binding on the spike glycoprotein of the severe acute respiratory syndrome coronavirus.
Auteurs : Yi-Ping Shih [République populaire de Chine] ; Chia-Yen Chen ; Shih-Jen Liu ; Kuan-Hsuan Chen ; Yuan-Ming Lee ; Yu-Chan Chao ; Yi-Ming Arthur ChenSource :
- Journal of virology [ 0022-538X ] ; 2006.
Descripteurs français
- KwdFr :
- ADN viral (génétique), Anticorps antiviraux, Anticorps monoclonaux, Antigènes viraux (génétique), Baculoviridae (génétique), Cartographie épitopique, Cellules dendritiques (immunologie), Cellules dendritiques (virologie), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (), Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (immunologie), Glycosylation, Humains, Lectines de type C (métabolisme), Liaison aux protéines, Modèles moléculaires, Molécules d'adhérence cellulaire (métabolisme), Mutagenèse dirigée, Protéines de l'enveloppe virale (), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (immunologie), Protéines recombinantes (), Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Récepteurs de surface cellulaire (métabolisme), Sites de fixation (génétique), Structure tertiaire des protéines, Syndrome respiratoire aigu sévère (immunologie), Syndrome respiratoire aigu sévère (virologie), Séquence nucléotidique, Techniques in vitro, Tests de neutralisation, Virus du SRAS (génétique), Virus du SRAS (immunologie), Épitopes (), Épitopes (génétique).
- MESH :
- génétique : ADN viral, Antigènes viraux, Baculoviridae, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes, Sites de fixation, Virus du SRAS, Épitopes.
- immunologie : Cellules dendritiques, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes, Syndrome respiratoire aigu sévère, Virus du SRAS.
- métabolisme : Lectines de type C, Molécules d'adhérence cellulaire, Récepteurs de surface cellulaire.
- virologie : Cellules dendritiques, Syndrome respiratoire aigu sévère.
- Anticorps antiviraux, Anticorps monoclonaux, Cartographie épitopique, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires, Glycosylation, Humains, Liaison aux protéines, Modèles moléculaires, Mutagenèse dirigée, Protéines de l'enveloppe virale, Protéines recombinantes, Structure tertiaire des protéines, Séquence nucléotidique, Techniques in vitro, Tests de neutralisation, Épitopes.
English descriptors
- KwdEn :
- Antibodies, Monoclonal, Antibodies, Viral, Antigens, Viral (genetics), Baculoviridae (genetics), Base Sequence, Binding Sites (genetics), Cell Adhesion Molecules (metabolism), DNA, Viral (genetics), Dendritic Cells (immunology), Dendritic Cells (virology), Epitope Mapping, Epitopes (chemistry), Epitopes (genetics), Glycosylation, Humans, In Vitro Techniques, Lectins, C-Type (metabolism), Membrane Glycoproteins (chemistry), Membrane Glycoproteins (genetics), Membrane Glycoproteins (immunology), Models, Molecular, Mutagenesis, Site-Directed, Neutralization Tests, Protein Binding, Protein Structure, Tertiary, Receptors, Cell Surface (metabolism), Recombinant Proteins (chemistry), Recombinant Proteins (genetics), Recombinant Proteins (immunology), SARS Virus (genetics), SARS Virus (immunology), Severe Acute Respiratory Syndrome (immunology), Severe Acute Respiratory Syndrome (virology), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (chemistry), Viral Envelope Proteins (genetics), Viral Envelope Proteins (immunology).
- MESH :
- chemical , chemistry : Epitopes, Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , genetics : Antigens, Viral, DNA, Viral, Epitopes, Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , immunology : Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , metabolism : Cell Adhesion Molecules, Lectins, C-Type, Receptors, Cell Surface.
- chemical : Antibodies, Monoclonal, Antibodies, Viral, Spike Glycoprotein, Coronavirus.
- genetics : Baculoviridae, Binding Sites, SARS Virus.
- immunology : Dendritic Cells, SARS Virus, Severe Acute Respiratory Syndrome.
- virology : Dendritic Cells, Severe Acute Respiratory Syndrome.
- Base Sequence, Epitope Mapping, Glycosylation, Humans, In Vitro Techniques, Models, Molecular, Mutagenesis, Site-Directed, Neutralization Tests, Protein Binding, Protein Structure, Tertiary.
Abstract
The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) uses dendritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN) to facilitate cell entry via cellular receptor-angiotensin-converting enzyme 2. For this project, we used recombinant baculoviruses expressing different lengths of SARS-CoV spike (S) protein in a capture assay to deduce the minimal DC-SIGN binding region. Our results identified the region location between amino acid (aa) residues 324 to 386 of the S protein. We then generated nine monoclonal antibodies (MAbs) against the S protein to map the DC-SIGN-binding domain using capture assays with pseudotyped viruses and observed that MAb SIa5 significantly blocked S protein-DC-SIGN interaction. An enhancement assay using the HKU39849 SARS-CoV strain and human immature dendritic cells confirmed our observation. Data from a pepscan analysis and M13 phage peptide display library system mapped the reactive MAb SIa5 epitope to aa residues 363 to 368 of the S protein. Results from a capture assay testing three pseudotyped viruses with mutated N-linked glycosylation sites of the S protein indicate that only two pseudotyped viruses (N330Q and N357Q, both of which lost glycosylation sites near the SIa5 epitope) had diminished DC-SIGN-binding capacity. We also noted that MAb SIb4 exerted a neutralizing effect against HKU39849; its reactive epitope was mapped to aa residues 435 to 439 of the S protein. We offer the data to facilitate the development of therapeutic agents and preventive vaccines against SARS-CoV infection.
DOI: 10.1128/JVI.01138-06
PubMed: 17041212
Affiliations:
Links toward previous steps (curation, corpus...)
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Links to Exploration step
pubmed:17041212Le document en format XML
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<author><name sortKey="Shih, Yi Ping" sort="Shih, Yi Ping" uniqKey="Shih Y" first="Yi-Ping" last="Shih">Yi-Ping Shih</name>
<affiliation wicri:level="1"><nlm:affiliation>AIDS Prevention and Research Center, National Yang-Ming University, Taipei 111, Taiwan, Republic of China.</nlm:affiliation>
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<author><name sortKey="Chen, Kuan Hsuan" sort="Chen, Kuan Hsuan" uniqKey="Chen K" first="Kuan-Hsuan" last="Chen">Kuan-Hsuan Chen</name>
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<author><name sortKey="Lee, Yuan Ming" sort="Lee, Yuan Ming" uniqKey="Lee Y" first="Yuan-Ming" last="Lee">Yuan-Ming Lee</name>
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<affiliation wicri:level="1"><nlm:affiliation>AIDS Prevention and Research Center, National Yang-Ming University, Taipei 111, Taiwan, Republic of China.</nlm:affiliation>
<country xml:lang="fr">République populaire de Chine</country>
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<author><name sortKey="Liu, Shih Jen" sort="Liu, Shih Jen" uniqKey="Liu S" first="Shih-Jen" last="Liu">Shih-Jen Liu</name>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Monoclonal</term>
<term>Antibodies, Viral</term>
<term>Antigens, Viral (genetics)</term>
<term>Baculoviridae (genetics)</term>
<term>Base Sequence</term>
<term>Binding Sites (genetics)</term>
<term>Cell Adhesion Molecules (metabolism)</term>
<term>DNA, Viral (genetics)</term>
<term>Dendritic Cells (immunology)</term>
<term>Dendritic Cells (virology)</term>
<term>Epitope Mapping</term>
<term>Epitopes (chemistry)</term>
<term>Epitopes (genetics)</term>
<term>Glycosylation</term>
<term>Humans</term>
<term>In Vitro Techniques</term>
<term>Lectins, C-Type (metabolism)</term>
<term>Membrane Glycoproteins (chemistry)</term>
<term>Membrane Glycoproteins (genetics)</term>
<term>Membrane Glycoproteins (immunology)</term>
<term>Models, Molecular</term>
<term>Mutagenesis, Site-Directed</term>
<term>Neutralization Tests</term>
<term>Protein Binding</term>
<term>Protein Structure, Tertiary</term>
<term>Receptors, Cell Surface (metabolism)</term>
<term>Recombinant Proteins (chemistry)</term>
<term>Recombinant Proteins (genetics)</term>
<term>Recombinant Proteins (immunology)</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (immunology)</term>
<term>Severe Acute Respiratory Syndrome (immunology)</term>
<term>Severe Acute Respiratory Syndrome (virology)</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Viral Envelope Proteins (chemistry)</term>
<term>Viral Envelope Proteins (genetics)</term>
<term>Viral Envelope Proteins (immunology)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ADN viral (génétique)</term>
<term>Anticorps antiviraux</term>
<term>Anticorps monoclonaux</term>
<term>Antigènes viraux (génétique)</term>
<term>Baculoviridae (génétique)</term>
<term>Cartographie épitopique</term>
<term>Cellules dendritiques (immunologie)</term>
<term>Cellules dendritiques (virologie)</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires ()</term>
<term>Glycoprotéines membranaires (génétique)</term>
<term>Glycoprotéines membranaires (immunologie)</term>
<term>Glycosylation</term>
<term>Humains</term>
<term>Lectines de type C (métabolisme)</term>
<term>Liaison aux protéines</term>
<term>Modèles moléculaires</term>
<term>Molécules d'adhérence cellulaire (métabolisme)</term>
<term>Mutagenèse dirigée</term>
<term>Protéines de l'enveloppe virale ()</term>
<term>Protéines de l'enveloppe virale (génétique)</term>
<term>Protéines de l'enveloppe virale (immunologie)</term>
<term>Protéines recombinantes ()</term>
<term>Protéines recombinantes (génétique)</term>
<term>Protéines recombinantes (immunologie)</term>
<term>Récepteurs de surface cellulaire (métabolisme)</term>
<term>Sites de fixation (génétique)</term>
<term>Structure tertiaire des protéines</term>
<term>Syndrome respiratoire aigu sévère (immunologie)</term>
<term>Syndrome respiratoire aigu sévère (virologie)</term>
<term>Séquence nucléotidique</term>
<term>Techniques in vitro</term>
<term>Tests de neutralisation</term>
<term>Virus du SRAS (génétique)</term>
<term>Virus du SRAS (immunologie)</term>
<term>Épitopes ()</term>
<term>Épitopes (génétique)</term>
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<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Epitopes</term>
<term>Membrane Glycoproteins</term>
<term>Recombinant Proteins</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antigens, Viral</term>
<term>DNA, Viral</term>
<term>Epitopes</term>
<term>Membrane Glycoproteins</term>
<term>Recombinant Proteins</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Membrane Glycoproteins</term>
<term>Recombinant Proteins</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Cell Adhesion Molecules</term>
<term>Lectins, C-Type</term>
<term>Receptors, Cell Surface</term>
</keywords>
<keywords scheme="MESH" type="chemical" xml:lang="en"><term>Antibodies, Monoclonal</term>
<term>Antibodies, Viral</term>
<term>Spike Glycoprotein, Coronavirus</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Baculoviridae</term>
<term>Binding Sites</term>
<term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN viral</term>
<term>Antigènes viraux</term>
<term>Baculoviridae</term>
<term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines recombinantes</term>
<term>Sites de fixation</term>
<term>Virus du SRAS</term>
<term>Épitopes</term>
</keywords>
<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Cellules dendritiques</term>
<term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines recombinantes</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Dendritic Cells</term>
<term>SARS Virus</term>
<term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Lectines de type C</term>
<term>Molécules d'adhérence cellulaire</term>
<term>Récepteurs de surface cellulaire</term>
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<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Cellules dendritiques</term>
<term>Syndrome respiratoire aigu sévère</term>
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<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Dendritic Cells</term>
<term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term>
<term>Epitope Mapping</term>
<term>Glycosylation</term>
<term>Humans</term>
<term>In Vitro Techniques</term>
<term>Models, Molecular</term>
<term>Mutagenesis, Site-Directed</term>
<term>Neutralization Tests</term>
<term>Protein Binding</term>
<term>Protein Structure, Tertiary</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Anticorps antiviraux</term>
<term>Anticorps monoclonaux</term>
<term>Cartographie épitopique</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires</term>
<term>Glycosylation</term>
<term>Humains</term>
<term>Liaison aux protéines</term>
<term>Modèles moléculaires</term>
<term>Mutagenèse dirigée</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines recombinantes</term>
<term>Structure tertiaire des protéines</term>
<term>Séquence nucléotidique</term>
<term>Techniques in vitro</term>
<term>Tests de neutralisation</term>
<term>Épitopes</term>
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<front><div type="abstract" xml:lang="en">The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) uses dendritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN) to facilitate cell entry via cellular receptor-angiotensin-converting enzyme 2. For this project, we used recombinant baculoviruses expressing different lengths of SARS-CoV spike (S) protein in a capture assay to deduce the minimal DC-SIGN binding region. Our results identified the region location between amino acid (aa) residues 324 to 386 of the S protein. We then generated nine monoclonal antibodies (MAbs) against the S protein to map the DC-SIGN-binding domain using capture assays with pseudotyped viruses and observed that MAb SIa5 significantly blocked S protein-DC-SIGN interaction. An enhancement assay using the HKU39849 SARS-CoV strain and human immature dendritic cells confirmed our observation. Data from a pepscan analysis and M13 phage peptide display library system mapped the reactive MAb SIa5 epitope to aa residues 363 to 368 of the S protein. Results from a capture assay testing three pseudotyped viruses with mutated N-linked glycosylation sites of the S protein indicate that only two pseudotyped viruses (N330Q and N357Q, both of which lost glycosylation sites near the SIa5 epitope) had diminished DC-SIGN-binding capacity. We also noted that MAb SIb4 exerted a neutralizing effect against HKU39849; its reactive epitope was mapped to aa residues 435 to 439 of the S protein. We offer the data to facilitate the development of therapeutic agents and preventive vaccines against SARS-CoV infection.</div>
</front>
</TEI>
<affiliations><list><country><li>République populaire de Chine</li>
</country>
</list>
<tree><noCountry><name sortKey="Chao, Yu Chan" sort="Chao, Yu Chan" uniqKey="Chao Y" first="Yu-Chan" last="Chao">Yu-Chan Chao</name>
<name sortKey="Chen, Chia Yen" sort="Chen, Chia Yen" uniqKey="Chen C" first="Chia-Yen" last="Chen">Chia-Yen Chen</name>
<name sortKey="Chen, Kuan Hsuan" sort="Chen, Kuan Hsuan" uniqKey="Chen K" first="Kuan-Hsuan" last="Chen">Kuan-Hsuan Chen</name>
<name sortKey="Chen, Yi Ming Arthur" sort="Chen, Yi Ming Arthur" uniqKey="Chen Y" first="Yi-Ming Arthur" last="Chen">Yi-Ming Arthur Chen</name>
<name sortKey="Lee, Yuan Ming" sort="Lee, Yuan Ming" uniqKey="Lee Y" first="Yuan-Ming" last="Lee">Yuan-Ming Lee</name>
<name sortKey="Liu, Shih Jen" sort="Liu, Shih Jen" uniqKey="Liu S" first="Shih-Jen" last="Liu">Shih-Jen Liu</name>
</noCountry>
<country name="République populaire de Chine"><noRegion><name sortKey="Shih, Yi Ping" sort="Shih, Yi Ping" uniqKey="Shih Y" first="Yi-Ping" last="Shih">Yi-Ping Shih</name>
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</record>
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