Identifying epitopes responsible for neutralizing antibody and DC-SIGN binding on the spike glycoprotein of the severe acute respiratory syndrome coronavirus
Identifieur interne : 004666 ( Main/Merge ); précédent : 004665; suivant : 004667Identifying epitopes responsible for neutralizing antibody and DC-SIGN binding on the spike glycoprotein of the severe acute respiratory syndrome coronavirus
Auteurs : Yi-Ping Shih [Taïwan] ; Chia-Yen Chen [Taïwan] ; Shih-Jen Liu [Taïwan] ; Kuan-Hsuan Chen [Taïwan] ; Yuan-Ming Lee [Taïwan] ; Yu-Chan Chao [Taïwan] ; Yi-Ming Arthur Chen [Taïwan]Source :
- Journal of virology [ 0022-538X ] ; 2006.
Descripteurs français
- Pascal (Inist)
English descriptors
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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.
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type="wicri:Area/Main/Merge">004666</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Identifying epitopes responsible for neutralizing antibody and DC-SIGN binding on the spike glycoprotein of the severe acute respiratory syndrome coronavirus</title><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"><inist:fA14 i1="01"><s1>AIDS Prevention and Research Center, National Yang- Ming University</s1><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>AIDS Prevention and Research Center, National Yang- Ming University</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Division of Preventive Medicine, Institute of Public Health, School of Medicine National Yang- Ming University</s1><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Division of Preventive Medicine, Institute of Public Health, School of Medicine National Yang- Ming University</wicri:noRegion></affiliation></author><author><name sortKey="Chen, Chia Yen" sort="Chen, Chia Yen" uniqKey="Chen C" first="Chia-Yen" last="Chen">Chia-Yen Chen</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>AIDS Prevention and Research Center, National Yang- Ming University</s1><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>AIDS Prevention and Research Center, National Yang- Ming University</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Division of Preventive Medicine, Institute of Public Health, School of Medicine National Yang- Ming University</s1><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Division of Preventive Medicine, Institute of Public Health, School of Medicine National Yang- Ming University</wicri:noRegion></affiliation></author><author><name sortKey="Liu, Shih Jen" sort="Liu, Shih Jen" uniqKey="Liu S" first="Shih-Jen" last="Liu">Shih-Jen Liu</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Vaccine Research and Development Center, National Health Research Institutes</s1><s3>TWN</s3><sZ>3 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Vaccine Research and Development Center, National Health Research Institutes</wicri:noRegion></affiliation></author><author><name sortKey="Chen, Kuan Hsuan" sort="Chen, Kuan Hsuan" uniqKey="Chen K" first="Kuan-Hsuan" last="Chen">Kuan-Hsuan Chen</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>AIDS Prevention and Research Center, National Yang- Ming University</s1><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>AIDS Prevention and Research Center, National Yang- Ming University</wicri:noRegion></affiliation></author><author><name sortKey="Lee, Yuan Ming" sort="Lee, Yuan Ming" uniqKey="Lee Y" first="Yuan-Ming" last="Lee">Yuan-Ming Lee</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Division of Preventive Medicine, Institute of Public Health, School of Medicine National Yang- Ming University</s1><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Division of Preventive Medicine, Institute of Public Health, School of Medicine National Yang- Ming University</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Pathology and Laboratory Diagnosis, Taipei Veterans General Hospital</s1><s3>TWN</s3><sZ>5 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Department of Pathology and Laboratory Diagnosis, Taipei Veterans General Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Chao, Yu Chan" sort="Chao, Yu Chan" uniqKey="Chao Y" first="Yu-Chan" last="Chao">Yu-Chan Chao</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Institute of Molecular Biology, Academia Sinica</s1><s2>Taipei</s2><s3>TWN</s3><sZ>6 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><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><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>AIDS Prevention and Research Center, National Yang- Ming University</s1><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>AIDS Prevention and Research Center, National Yang- Ming University</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Division of Preventive Medicine, Institute of Public Health, School of Medicine National Yang- Ming University</s1><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Division of Preventive Medicine, Institute of Public Health, School of Medicine National Yang- Ming University</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno><imprint><date when="2006">2006</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antigenic determinant</term><term>Coronavirus</term><term>Glycoprotein</term><term>Microbiology</term><term>Neutralizing antibody</term><term>Severe acute respiratory syndrome</term><term>Virology</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term><term>Déterminant antigénique</term><term>Anticorps neutralisant</term><term>Glycoprotéine</term><term>Microbiologie</term><term>Virologie</term><term>Syndrome respiratoire aigu sévère</term></keywords></textClass></profileDesc></teiHeader><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>Taïwan</li></country></list><tree><country name="Taïwan"><noRegion><name sortKey="Shih, Yi Ping" sort="Shih, Yi Ping" uniqKey="Shih Y" first="Yi-Ping" last="Shih">Yi-Ping Shih</name></noRegion><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, 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="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="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><name sortKey="Shih, Yi Ping" sort="Shih, Yi Ping" uniqKey="Shih Y" first="Yi-Ping" last="Shih">Yi-Ping Shih</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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