Amino acids 1055 to 1192 in the S2 region of severe acute respiratory syndrome coronavirus S protein induce neutralizing antibodies : Implications for the development of vaccines and antiviral agents
Identifieur interne : 000713 ( PascalFrancis/Checkpoint ); précédent : 000712; suivant : 000714Amino acids 1055 to 1192 in the S2 region of severe acute respiratory syndrome coronavirus S protein induce neutralizing antibodies : Implications for the development of vaccines and antiviral agents
Auteurs : Choong-Tat Keng [Singapour] ; AIHUA ZHANG [République populaire de Chine] ; SHUO SHEN [Singapour] ; Kuo-Ming Lip [Singapour] ; Burtram C. Fielding [Singapour] ; Timothy H. P. Tan [Singapour] ; Chih-Fong Chou [Singapour] ; CHAY BOON LOH [Singapour] ; SIFANG WANG [Singapour] ; JIANLIN FU [Singapour] ; XIAOMING YANG [République populaire de Chine] ; SENG GEE LIM [Singapour] ; WANJIN HONG [Singapour] ; Yee-Joo Tan [Singapour]Source :
- Journal of virology [ 0022-538X ] ; 2005.
Descripteurs français
- Pascal (Inist)
- Wicri :
- topic : Vaccin.
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Abstract
The spike (S) protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) interacts with cellular receptors to mediate membrane fusion, allowing viral entry into host cells; hence it is recognized as the primary target of neutralizing antibodies, and therefore knowledge of antigenic determinants that can elicit neutralizing antibodies could be beneficial for the development of a protective vaccine. Here, we expressed five different fragments of S, covering the entire ectodomain (amino acids 48 to 1192), as glutathione S-transferase fusion proteins in Escherichia coli and used the purified proteins to raise antibodies in rabbits. By Western blot analysis and immunoprecipitation experiments, we showed that all the antibodies are specific and highly sensitive to both the native and denatured forms of the full-length S protein expressed in virus-infected cells and transfected cells, respectively. Indirect immunofluorescence performed on fixed but unpermeabilized cells showed that these antibodies can recognize the mature form of S on the cell surface. All the antibodies were also able to detect the maturation of the 200-kDa form of S to the 210-kDa form by pulse-chase experiments. When the antibodies were tested for their ability to inhibit SARS-CoV propagation in Vero E6 culture, it was found that the anti-SΔ10 antibody, which was targeted to amino acid residues 1029 to 1192 of S, which include heptad repeat 2, has strong neutralizing activities, suggesting that this region of S carries neutralizing epitopes and is very important for virus entry into cells.
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Fielding</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Tan, Timothy H P" sort="Tan, Timothy H P" uniqKey="Tan T" first="Timothy H. P." last="Tan">Timothy H. P. Tan</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Chou, Chih Fong" sort="Chou, Chih Fong" uniqKey="Chou C" first="Chih-Fong" last="Chou">Chih-Fong Chou</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Chay Boon Loh" sort="Chay Boon Loh" uniqKey="Chay Boon Loh" last="Chay Boon Loh">CHAY BOON LOH</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Sifang Wang" sort="Sifang Wang" uniqKey="Sifang Wang" last="Sifang Wang">SIFANG WANG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Jianlin Fu" sort="Jianlin Fu" uniqKey="Jianlin Fu" last="Jianlin Fu">JIANLIN FU</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Xiaoming Yang" sort="Xiaoming Yang" uniqKey="Xiaoming Yang" last="Xiaoming Yang">XIAOMING YANG</name><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Wuhan Institute of Biological Products</s1><s2>Wuhan</s2><s3>CHN</s3><sZ>2 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation></author><author><name sortKey="Seng Gee Lim" sort="Seng Gee Lim" uniqKey="Seng Gee Lim" last="Seng Gee Lim">SENG GEE LIM</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Wanjin Hong" sort="Wanjin Hong" uniqKey="Wanjin Hong" last="Wanjin Hong">WANJIN HONG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Tan, Yee Joo" sort="Tan, Yee Joo" uniqKey="Tan Y" first="Yee-Joo" last="Tan">Yee-Joo Tan</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">05-0158491</idno><date when="2005">2005</date><idno type="stanalyst">PASCAL 05-0158491 INIST</idno><idno type="RBID">Pascal:05-0158491</idno><idno type="wicri:Area/PascalFrancis/Corpus">000712</idno><idno type="wicri:Area/PascalFrancis/Curation">000278</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000713</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000713</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Amino acids 1055 to 1192 in the S2 region of severe acute respiratory syndrome coronavirus S protein induce neutralizing antibodies : Implications for the development of vaccines and antiviral agents</title><author><name sortKey="Keng, Choong Tat" sort="Keng, Choong Tat" uniqKey="Keng C" first="Choong-Tat" last="Keng">Choong-Tat Keng</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Aihua Zhang" sort="Aihua Zhang" uniqKey="Aihua Zhang" last="Aihua Zhang">AIHUA ZHANG</name><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Wuhan Institute of Biological Products</s1><s2>Wuhan</s2><s3>CHN</s3><sZ>2 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation></author><author><name sortKey="Shuo Shen" sort="Shuo Shen" uniqKey="Shuo Shen" last="Shuo Shen">SHUO SHEN</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Lip, Kuo Ming" sort="Lip, Kuo Ming" uniqKey="Lip K" first="Kuo-Ming" last="Lip">Kuo-Ming Lip</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Fielding, Burtram C" sort="Fielding, Burtram C" uniqKey="Fielding B" first="Burtram C." last="Fielding">Burtram C. Fielding</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Tan, Timothy H P" sort="Tan, Timothy H P" uniqKey="Tan T" first="Timothy H. P." last="Tan">Timothy H. P. Tan</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Chou, Chih Fong" sort="Chou, Chih Fong" uniqKey="Chou C" first="Chih-Fong" last="Chou">Chih-Fong Chou</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Chay Boon Loh" sort="Chay Boon Loh" uniqKey="Chay Boon Loh" last="Chay Boon Loh">CHAY BOON LOH</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Sifang Wang" sort="Sifang Wang" uniqKey="Sifang Wang" last="Sifang Wang">SIFANG WANG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Jianlin Fu" sort="Jianlin Fu" uniqKey="Jianlin Fu" last="Jianlin Fu">JIANLIN FU</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Xiaoming Yang" sort="Xiaoming Yang" uniqKey="Xiaoming Yang" last="Xiaoming Yang">XIAOMING YANG</name><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Wuhan Institute of Biological Products</s1><s2>Wuhan</s2><s3>CHN</s3><sZ>2 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation></author><author><name sortKey="Seng Gee Lim" sort="Seng Gee Lim" uniqKey="Seng Gee Lim" last="Seng Gee Lim">SENG GEE LIM</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Wanjin Hong" sort="Wanjin Hong" uniqKey="Wanjin Hong" last="Wanjin Hong">WANJIN HONG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</wicri:noRegion></affiliation></author><author><name sortKey="Tan, Yee Joo" sort="Tan, Yee Joo" uniqKey="Tan Y" first="Yee-Joo" last="Tan">Yee-Joo Tan</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>Institute of Molecular and Cell Biology</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="2005">2005</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>Antiviral</term><term>Coronavirus</term><term>Microbiology</term><term>Neutralizing antibody</term><term>Protein</term><term>Severe acute respiratory syndrome</term><term>Vaccine</term><term>Virology</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term><term>Protéine</term><term>Anticorps neutralisant</term><term>Vaccin</term><term>Antiviral</term><term>Microbiologie</term><term>Virologie</term><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Vaccin</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The spike (S) protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) interacts with cellular receptors to mediate membrane fusion, allowing viral entry into host cells; hence it is recognized as the primary target of neutralizing antibodies, and therefore knowledge of antigenic determinants that can elicit neutralizing antibodies could be beneficial for the development of a protective vaccine. Here, we expressed five different fragments of S, covering the entire ectodomain (amino acids 48 to 1192), as glutathione S-transferase fusion proteins in Escherichia coli and used the purified proteins to raise antibodies in rabbits. By Western blot analysis and immunoprecipitation experiments, we showed that all the antibodies are specific and highly sensitive to both the native and denatured forms of the full-length S protein expressed in virus-infected cells and transfected cells, respectively. Indirect immunofluorescence performed on fixed but unpermeabilized cells showed that these antibodies can recognize the mature form of S on the cell surface. All the antibodies were also able to detect the maturation of the 200-kDa form of S to the 210-kDa form by pulse-chase experiments. When the antibodies were tested for their ability to inhibit SARS-CoV propagation in Vero E6 culture, it was found that the anti-SΔ10 antibody, which was targeted to amino acid residues 1029 to 1192 of S, which include heptad repeat 2, has strong neutralizing activities, suggesting that this region of S carries neutralizing epitopes and is very important for virus entry into cells.</div></front></TEI><inist><standard h6="B"><pA><fA01 i1="01" i2="1"><s0>0022-538X</s0></fA01><fA03 i2="1"><s0>J. virol.</s0></fA03><fA05><s2>79</s2></fA05><fA06><s2>6</s2></fA06><fA08 i1="01" i2="1" l="ENG"><s1>Amino acids 1055 to 1192 in the S2 region of severe acute respiratory syndrome coronavirus S protein induce neutralizing antibodies : Implications for the development of vaccines and antiviral agents</s1></fA08><fA11 i1="01" i2="1"><s1>KENG (Choong-Tat)</s1></fA11><fA11 i1="02" i2="1"><s1>AIHUA ZHANG</s1></fA11><fA11 i1="03" i2="1"><s1>SHUO SHEN</s1></fA11><fA11 i1="04" i2="1"><s1>LIP (Kuo-Ming)</s1></fA11><fA11 i1="05" i2="1"><s1>FIELDING (Burtram C.)</s1></fA11><fA11 i1="06" i2="1"><s1>TAN (Timothy H. P.)</s1></fA11><fA11 i1="07" i2="1"><s1>CHOU (Chih-Fong)</s1></fA11><fA11 i1="08" i2="1"><s1>CHAY BOON LOH</s1></fA11><fA11 i1="09" i2="1"><s1>SIFANG WANG</s1></fA11><fA11 i1="10" i2="1"><s1>JIANLIN FU</s1></fA11><fA11 i1="11" i2="1"><s1>XIAOMING YANG</s1></fA11><fA11 i1="12" i2="1"><s1>SENG GEE LIM</s1></fA11><fA11 i1="13" i2="1"><s1>WANJIN HONG</s1></fA11><fA11 i1="14" i2="1"><s1>TAN (Yee-Joo)</s1></fA11><fA14 i1="01"><s1>Institute of Molecular and Cell Biology</s1><s3>SGP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ></fA14><fA14 i1="02"><s1>Wuhan Institute of Biological Products</s1><s2>Wuhan</s2><s3>CHN</s3><sZ>2 aut.</sZ><sZ>11 aut.</sZ></fA14><fA20><s1>3289-3296</s1></fA20><fA21><s1>2005</s1></fA21><fA23 i1="01"><s0>ENG</s0></fA23><fA43 i1="01"><s1>INIST</s1><s2>13592</s2><s5>354000126300760060</s5></fA43><fA44><s0>0000</s0><s1>© 2005 INIST-CNRS. All rights reserved.</s1></fA44><fA45><s0>38 ref.</s0></fA45><fA47 i1="01" i2="1"><s0>05-0158491</s0></fA47><fA60><s1>P</s1></fA60><fA61><s0>A</s0></fA61><fA64 i1="01" i2="1"><s0>Journal of virology</s0></fA64><fA66 i1="01"><s0>USA</s0></fA66><fC01 i1="01" l="ENG"><s0>The spike (S) protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) interacts with cellular receptors to mediate membrane fusion, allowing viral entry into host cells; hence it is recognized as the primary target of neutralizing antibodies, and therefore knowledge of antigenic determinants that can elicit neutralizing antibodies could be beneficial for the development of a protective vaccine. Here, we expressed five different fragments of S, covering the entire ectodomain (amino acids 48 to 1192), as glutathione S-transferase fusion proteins in Escherichia coli and used the purified proteins to raise antibodies in rabbits. By Western blot analysis and immunoprecipitation experiments, we showed that all the antibodies are specific and highly sensitive to both the native and denatured forms of the full-length S protein expressed in virus-infected cells and transfected cells, respectively. Indirect immunofluorescence performed on fixed but unpermeabilized cells showed that these antibodies can recognize the mature form of S on the cell surface. All the antibodies were also able to detect the maturation of the 200-kDa form of S to the 210-kDa form by pulse-chase experiments. When the antibodies were tested for their ability to inhibit SARS-CoV propagation in Vero E6 culture, it was found that the anti-SΔ10 antibody, which was targeted to amino acid residues 1029 to 1192 of S, which include heptad repeat 2, has strong neutralizing activities, suggesting that this region of S carries neutralizing epitopes and is very important for virus entry into cells.</s0></fC01><fC02 i1="01" i2="X"><s0>002A05C10</s0></fC02><fC02 i1="02" i2="X"><s0>002A05C07</s0></fC02><fC03 i1="01" i2="X" l="FRE"><s0>Coronavirus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="01" i2="X" l="ENG"><s0>Coronavirus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="01" i2="X" l="SPA"><s0>Coronavirus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="02" i2="X" l="FRE"><s0>Protéine</s0><s5>05</s5></fC03><fC03 i1="02" i2="X" l="ENG"><s0>Protein</s0><s5>05</s5></fC03><fC03 i1="02" i2="X" l="SPA"><s0>Proteína</s0><s5>05</s5></fC03><fC03 i1="03" i2="X" l="FRE"><s0>Anticorps neutralisant</s0><s5>06</s5></fC03><fC03 i1="03" i2="X" l="ENG"><s0>Neutralizing antibody</s0><s5>06</s5></fC03><fC03 i1="03" i2="X" l="SPA"><s0>anticuerpo neutralizante</s0><s5>06</s5></fC03><fC03 i1="04" i2="X" l="FRE"><s0>Vaccin</s0><s5>07</s5></fC03><fC03 i1="04" i2="X" l="ENG"><s0>Vaccine</s0><s5>07</s5></fC03><fC03 i1="04" i2="X" l="SPA"><s0>Vacuna</s0><s5>07</s5></fC03><fC03 i1="05" i2="X" l="FRE"><s0>Antiviral</s0><s5>08</s5></fC03><fC03 i1="05" i2="X" l="ENG"><s0>Antiviral</s0><s5>08</s5></fC03><fC03 i1="05" i2="X" l="SPA"><s0>Antiviral</s0><s5>08</s5></fC03><fC03 i1="06" i2="X" l="FRE"><s0>Microbiologie</s0><s5>09</s5></fC03><fC03 i1="06" i2="X" l="ENG"><s0>Microbiology</s0><s5>09</s5></fC03><fC03 i1="06" i2="X" l="SPA"><s0>Microbiología</s0><s5>09</s5></fC03><fC03 i1="07" i2="X" l="FRE"><s0>Virologie</s0><s5>10</s5></fC03><fC03 i1="07" i2="X" l="ENG"><s0>Virology</s0><s5>10</s5></fC03><fC03 i1="07" i2="X" l="SPA"><s0>Virología</s0><s5>10</s5></fC03><fC03 i1="08" i2="X" l="FRE"><s0>Syndrome respiratoire aigu sévère</s0><s2>NM</s2><s5>14</s5></fC03><fC03 i1="08" i2="X" l="ENG"><s0>Severe acute respiratory syndrome</s0><s2>NM</s2><s5>14</s5></fC03><fC03 i1="08" i2="X" l="SPA"><s0>Síndrome respiratorio agudo severo</s0><s2>NM</s2><s5>14</s5></fC03><fC07 i1="01" i2="X" l="FRE"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="ENG"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="SPA"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="FRE"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="ENG"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="SPA"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="FRE"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="ENG"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="SPA"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="04" i2="X" l="FRE"><s0>Appareil respiratoire pathologie</s0><s5>13</s5></fC07><fC07 i1="04" i2="X" l="ENG"><s0>Respiratory disease</s0><s5>13</s5></fC07><fC07 i1="04" i2="X" l="SPA"><s0>Aparato respiratorio patología</s0><s5>13</s5></fC07><fC07 i1="05" i2="X" l="FRE"><s0>Virose</s0></fC07><fC07 i1="05" i2="X" l="ENG"><s0>Viral disease</s0></fC07><fC07 i1="05" i2="X" l="SPA"><s0>Virosis</s0></fC07><fC07 i1="06" i2="X" l="FRE"><s0>Infection</s0></fC07><fC07 i1="06" i2="X" l="ENG"><s0>Infection</s0></fC07><fC07 i1="06" i2="X" l="SPA"><s0>Infección</s0></fC07><fC07 i1="07" i2="X" l="FRE"><s0>Poumon pathologie</s0><s5>16</s5></fC07><fC07 i1="07" i2="X" l="ENG"><s0>Lung disease</s0><s5>16</s5></fC07><fC07 i1="07" i2="X" l="SPA"><s0>Pulmón patología</s0><s5>16</s5></fC07><fN21><s1>108</s1></fN21><fN44 i1="01"><s1>OTO</s1></fN44><fN82><s1>OTO</s1></fN82></pA></standard></inist><affiliations><list><country><li>République populaire de Chine</li><li>Singapour</li></country><region><li>Hubei</li></region><settlement><li>Wuhan</li></settlement></list><tree><country name="Singapour"><noRegion><name sortKey="Keng, Choong Tat" sort="Keng, Choong Tat" uniqKey="Keng C" first="Choong-Tat" last="Keng">Choong-Tat Keng</name></noRegion><name sortKey="Chay Boon Loh" sort="Chay Boon Loh" uniqKey="Chay Boon Loh" last="Chay Boon Loh">CHAY BOON LOH</name><name sortKey="Chou, Chih Fong" sort="Chou, Chih Fong" uniqKey="Chou C" first="Chih-Fong" last="Chou">Chih-Fong Chou</name><name sortKey="Fielding, Burtram C" sort="Fielding, Burtram C" uniqKey="Fielding B" first="Burtram C." last="Fielding">Burtram C. Fielding</name><name sortKey="Jianlin Fu" sort="Jianlin Fu" uniqKey="Jianlin Fu" last="Jianlin Fu">JIANLIN FU</name><name sortKey="Lip, Kuo Ming" sort="Lip, Kuo Ming" uniqKey="Lip K" first="Kuo-Ming" last="Lip">Kuo-Ming Lip</name><name sortKey="Seng Gee Lim" sort="Seng Gee Lim" uniqKey="Seng Gee Lim" last="Seng Gee Lim">SENG GEE LIM</name><name sortKey="Shuo Shen" sort="Shuo Shen" uniqKey="Shuo Shen" last="Shuo Shen">SHUO SHEN</name><name sortKey="Sifang Wang" sort="Sifang Wang" uniqKey="Sifang Wang" last="Sifang Wang">SIFANG WANG</name><name sortKey="Tan, Timothy H P" sort="Tan, Timothy H P" uniqKey="Tan T" first="Timothy H. P." last="Tan">Timothy H. P. Tan</name><name sortKey="Tan, Yee Joo" sort="Tan, Yee Joo" uniqKey="Tan Y" first="Yee-Joo" last="Tan">Yee-Joo Tan</name><name sortKey="Wanjin Hong" sort="Wanjin Hong" uniqKey="Wanjin Hong" last="Wanjin Hong">WANJIN HONG</name></country><country name="République populaire de Chine"><region name="Hubei"><name sortKey="Aihua Zhang" sort="Aihua Zhang" uniqKey="Aihua Zhang" last="Aihua Zhang">AIHUA ZHANG</name></region><name sortKey="Xiaoming Yang" sort="Xiaoming Yang" uniqKey="Xiaoming Yang" last="Xiaoming Yang">XIAOMING YANG</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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