[Development and application of a safe SARS-CoV neutralization assay based on lentiviral vectors pseudotyped with SARS-CoV spike protein].
Identifieur interne : 003545 ( Main/Exploration ); précédent : 003544; suivant : 003546[Development and application of a safe SARS-CoV neutralization assay based on lentiviral vectors pseudotyped with SARS-CoV spike protein].
Auteurs : Ke-Xia Yan [République populaire de Chine] ; Wen-Jie Tan ; Xiang-Min Zhang ; Hui-Juan Wang ; Yan Li ; Li RuanSource :
- Bing du xue bao = Chinese journal of virology [ 1000-8721 ] ; 2007.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (sang), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (immunologie), Lentivirus (génétique), Plan de recherche, Plasmides, Protéines de l'enveloppe virale (immunologie), Protéines recombinantes (immunologie), Souris, Technique de Western, Tests de neutralisation (), Virus du SRAS (immunologie).
- MESH :
- génétique : Lentivirus.
- immunologie : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes, Virus du SRAS.
- sang : Anticorps antiviraux.
- Animaux, Glycoprotéine de spicule des coronavirus, Plan de recherche, Plasmides, Souris, Technique de Western, Tests de neutralisation.
English descriptors
- KwdEn :
- Animals, Antibodies, Viral (blood), Blotting, Western, Lentivirus (genetics), Membrane Glycoproteins (immunology), Mice, Neutralization Tests (methods), Plasmids, Recombinant Proteins (immunology), Research Design, SARS Virus (immunology), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (immunology).
- MESH :
- chemical , blood : Antibodies, Viral.
- genetics : Lentivirus.
- chemical , immunology : Membrane Glycoproteins, Recombinant Proteins, SARS Virus, Viral Envelope Proteins.
- methods : Neutralization Tests.
- Animals, Blotting, Western, Mice, Plasmids, Research Design, Spike Glycoprotein, Coronavirus.
Abstract
The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike protein (S) is a major target for neutralizing antibody. To develop and apply a safe neutralization assay for SARS-CoV, lentiviral SARS-CoV S pseudotypes had been constructed based on a three plasmid system, which contained pVRC8304 (harboring codon optimized full-length SARS-CoV S protein), pCMV delta 8. 2 (HIV-1 gag/pol construct) and pHR'CMV EGFP (the green fluorescent protein reporter construct). The pseudo-typed lentiviral particles were used to develop an in vitro microneutralization assay that was both sensitive and specific for SARS-CoV neutralizing antibody. We used this assay to determine the titers of the neutralizing antibodies (Nabs) in serum samples from mice immunized with various rVVs expressing different S fragments of SARS-CoV. The serum antibodies derived from S and various segments of S1 region neutralized SARS-CoV in vitro. No cross-neutralization occurred with the goat antiserum prepared with inactivated HCoV-OC43 or HCoV-229E. Neutralization titers measured by this assay were highly parallel with those measured by the assay using live SARS-CoV. Because the pseudotype assay does not require handling live SARS virus, it is a useful tool to determine serum neutralizing titers during natural infection and the preclinical evaluation of candidate vaccines.
PubMed: 18092680
Affiliations:
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China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Tan, Wen Jie" sort="Tan, Wen Jie" uniqKey="Tan W" first="Wen-Jie" last="Tan">Wen-Jie Tan</name></author><author><name sortKey="Zhang, Xiang Min" sort="Zhang, Xiang Min" uniqKey="Zhang X" first="Xiang-Min" last="Zhang">Xiang-Min Zhang</name></author><author><name sortKey="Wang, Hui Juan" sort="Wang, Hui Juan" uniqKey="Wang H" first="Hui-Juan" last="Wang">Hui-Juan Wang</name></author><author><name sortKey="Li, Yan" sort="Li, Yan" uniqKey="Li Y" first="Yan" last="Li">Yan Li</name></author><author><name sortKey="Ruan, Li" sort="Ruan, Li" uniqKey="Ruan L" first="Li" last="Ruan">Li Ruan</name></author></analytic><series><title level="j">Bing du xue bao = Chinese journal of virology</title><idno type="ISSN">1000-8721</idno><imprint><date when="2007" type="published">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Viral (blood)</term><term>Blotting, Western</term><term>Lentivirus (genetics)</term><term>Membrane Glycoproteins (immunology)</term><term>Mice</term><term>Neutralization Tests (methods)</term><term>Plasmids</term><term>Recombinant Proteins (immunology)</term><term>Research Design</term><term>SARS Virus (immunology)</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps antiviraux (sang)</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (immunologie)</term><term>Lentivirus (génétique)</term><term>Plan de recherche</term><term>Plasmides</term><term>Protéines de l'enveloppe virale (immunologie)</term><term>Protéines recombinantes (immunologie)</term><term>Souris</term><term>Technique de Western</term><term>Tests de neutralisation ()</term><term>Virus du SRAS (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antibodies, Viral</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Lentivirus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Lentivirus</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Protéines recombinantes</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Membrane Glycoproteins</term><term>Recombinant Proteins</term><term>SARS Virus</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Neutralization Tests</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>Anticorps antiviraux</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Blotting, Western</term><term>Mice</term><term>Plasmids</term><term>Research Design</term><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Glycoprotéine de spicule des coronavirus</term><term>Plan de recherche</term><term>Plasmides</term><term>Souris</term><term>Technique de Western</term><term>Tests de neutralisation</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike protein (S) is a major target for neutralizing antibody. To develop and apply a safe neutralization assay for SARS-CoV, lentiviral SARS-CoV S pseudotypes had been constructed based on a three plasmid system, which contained pVRC8304 (harboring codon optimized full-length SARS-CoV S protein), pCMV delta 8. 2 (HIV-1 gag/pol construct) and pHR'CMV EGFP (the green fluorescent protein reporter construct). The pseudo-typed lentiviral particles were used to develop an in vitro microneutralization assay that was both sensitive and specific for SARS-CoV neutralizing antibody. We used this assay to determine the titers of the neutralizing antibodies (Nabs) in serum samples from mice immunized with various rVVs expressing different S fragments of SARS-CoV. The serum antibodies derived from S and various segments of S1 region neutralized SARS-CoV in vitro. No cross-neutralization occurred with the goat antiserum prepared with inactivated HCoV-OC43 or HCoV-229E. Neutralization titers measured by this assay were highly parallel with those measured by the assay using live SARS-CoV. Because the pseudotype assay does not require handling live SARS virus, it is a useful tool to determine serum neutralizing titers during natural infection and the preclinical evaluation of candidate vaccines.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Li, Yan" sort="Li, Yan" uniqKey="Li Y" first="Yan" last="Li">Yan Li</name><name sortKey="Ruan, Li" sort="Ruan, Li" uniqKey="Ruan L" first="Li" last="Ruan">Li Ruan</name><name sortKey="Tan, Wen Jie" sort="Tan, Wen Jie" uniqKey="Tan W" first="Wen-Jie" last="Tan">Wen-Jie Tan</name><name sortKey="Wang, Hui Juan" sort="Wang, Hui Juan" uniqKey="Wang H" first="Hui-Juan" last="Wang">Hui-Juan Wang</name><name sortKey="Zhang, Xiang Min" sort="Zhang, Xiang Min" uniqKey="Zhang X" first="Xiang-Min" last="Zhang">Xiang-Min Zhang</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Yan, Ke Xia" sort="Yan, Ke Xia" uniqKey="Yan K" first="Ke-Xia" last="Yan">Ke-Xia Yan</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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