A convenient cell fusion assay for the study of SARS-CoV entry and inhibition.
Identifieur interne : 002333 ( PubMed/Checkpoint ); précédent : 002332; suivant : 002334A convenient cell fusion assay for the study of SARS-CoV entry and inhibition.
Auteurs : Yonggang Sha [République populaire de Chine] ; Yingliang Wu ; Zhijian Cao ; Xiuling Xu ; Wenlan Wu ; Dahe Jiang ; Xin Mao ; Hui Liu ; Ying Zhu ; Rui Gong ; Wenxin LiSource :
- IUBMB life [ 1521-6543 ] ; 2006.
Descripteurs français
- KwdFr :
- Animaux, Cellules COS, Colorants fluorescents (métabolisme), Dosage biologique, Escherichia coli (génétique), Facteurs temps, Fusion cellulaire, Glutathione transferase (métabolisme), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (métabolisme), Peptides (), Peptides (métabolisme), Peptidyl-Dipeptidase A (métabolisme), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (métabolisme), Protéines luminescentes (métabolisme), Protéines recombinantes (métabolisme), Protéines à fluorescence verte (métabolisme), Syndrome respiratoire aigu sévère (métabolisme), Techniques de coculture, Trypsine (pharmacologie), Virus du SRAS (génétique), Virus du SRAS (métabolisme).
- MESH :
- génétique : Escherichia coli, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Virus du SRAS.
- métabolisme : Colorants fluorescents, Glutathione transferase, Glycoprotéines membranaires, Peptides, Peptidyl-Dipeptidase A, Protéines de l'enveloppe virale, Protéines luminescentes, Protéines recombinantes, Protéines à fluorescence verte, Syndrome respiratoire aigu sévère, Virus du SRAS.
- pharmacologie : Trypsine.
- Animaux, Cellules COS, Dosage biologique, Facteurs temps, Fusion cellulaire, Glycoprotéine de spicule des coronavirus, Peptides, Techniques de coculture.
English descriptors
- KwdEn :
- Animals, Biological Assay, COS Cells, Cell Fusion, Chlorocebus aethiops, Coculture Techniques, Escherichia coli (genetics), Fluorescent Dyes (metabolism), Glutathione Transferase (metabolism), Green Fluorescent Proteins (metabolism), Luminescent Proteins (metabolism), Membrane Glycoproteins (genetics), Membrane Glycoproteins (metabolism), Peptides (chemistry), Peptides (metabolism), Peptidyl-Dipeptidase A (metabolism), Recombinant Proteins (metabolism), SARS Virus (genetics), SARS Virus (metabolism), Severe Acute Respiratory Syndrome (metabolism), Spike Glycoprotein, Coronavirus, Time Factors, Trypsin (pharmacology), Viral Envelope Proteins (genetics), Viral Envelope Proteins (metabolism).
- MESH :
- chemical , chemistry : Peptides.
- chemical , genetics : Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , metabolism : Fluorescent Dyes, Glutathione Transferase, Green Fluorescent Proteins, Luminescent Proteins, Membrane Glycoproteins, Peptides, Peptidyl-Dipeptidase A, Recombinant Proteins, Viral Envelope Proteins.
- genetics : Escherichia coli, SARS Virus.
- metabolism : SARS Virus, Severe Acute Respiratory Syndrome.
- chemical , pharmacology : Trypsin.
- Animals, Biological Assay, COS Cells, Cell Fusion, Chlorocebus aethiops, Coculture Techniques, Spike Glycoprotein, Coronavirus, Time Factors.
Abstract
SARS-CoV spike (S) protein-mediated cell fusion is important for the viral entry mechanism and identification of SARS-CoV entry inhibitors. In order to avoid the high risks involved in handling SARS-CoV and to facilitate the study of viral fusion mechanism, we established the cell lines: SR-COS7 cells that stably express both SARS-CoV S protein and red fluorescence protein, R-COS7 cells that stably express red fluorescence protein, and AG-COS7 cells that stably express both ACE2 and green fluorescence protein, respectively. When SR-COS7 cells or R-COS7 cells were cocultured with AG-COS7 cells, syncytia with yellow fluorescence were conveniently observed after 12 h in SR-COS7 cells plus AG-COS7 cells, but not in R-COS7 cells plus AG-COS7 cells. The cell-to-cell fusion efficiency was simply determined for quantitative analysis based on the number of syncytium detected by flow cytometry. Such new cell-to-cell fusion model was further assessed by the potent HR2 peptide inhibitor, which led to the obvious decrease of the cell-to-cell fusion efficiency. The successful fusion and inhibition of cell-based binding assay shows that it can be well used for the study of SARS-CoV entry and inhibition.
DOI: 10.1080/15216540600820974
PubMed: 16916786
Affiliations:
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pubmed:16916786Le document en format XML
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<front><div type="abstract" xml:lang="en">SARS-CoV spike (S) protein-mediated cell fusion is important for the viral entry mechanism and identification of SARS-CoV entry inhibitors. In order to avoid the high risks involved in handling SARS-CoV and to facilitate the study of viral fusion mechanism, we established the cell lines: SR-COS7 cells that stably express both SARS-CoV S protein and red fluorescence protein, R-COS7 cells that stably express red fluorescence protein, and AG-COS7 cells that stably express both ACE2 and green fluorescence protein, respectively. When SR-COS7 cells or R-COS7 cells were cocultured with AG-COS7 cells, syncytia with yellow fluorescence were conveniently observed after 12 h in SR-COS7 cells plus AG-COS7 cells, but not in R-COS7 cells plus AG-COS7 cells. The cell-to-cell fusion efficiency was simply determined for quantitative analysis based on the number of syncytium detected by flow cytometry. Such new cell-to-cell fusion model was further assessed by the potent HR2 peptide inhibitor, which led to the obvious decrease of the cell-to-cell fusion efficiency. The successful fusion and inhibition of cell-based binding assay shows that it can be well used for the study of SARS-CoV entry and inhibition.</div>
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<Abstract><AbstractText>SARS-CoV spike (S) protein-mediated cell fusion is important for the viral entry mechanism and identification of SARS-CoV entry inhibitors. In order to avoid the high risks involved in handling SARS-CoV and to facilitate the study of viral fusion mechanism, we established the cell lines: SR-COS7 cells that stably express both SARS-CoV S protein and red fluorescence protein, R-COS7 cells that stably express red fluorescence protein, and AG-COS7 cells that stably express both ACE2 and green fluorescence protein, respectively. When SR-COS7 cells or R-COS7 cells were cocultured with AG-COS7 cells, syncytia with yellow fluorescence were conveniently observed after 12 h in SR-COS7 cells plus AG-COS7 cells, but not in R-COS7 cells plus AG-COS7 cells. The cell-to-cell fusion efficiency was simply determined for quantitative analysis based on the number of syncytium detected by flow cytometry. Such new cell-to-cell fusion model was further assessed by the potent HR2 peptide inhibitor, which led to the obvious decrease of the cell-to-cell fusion efficiency. The successful fusion and inhibition of cell-based binding assay shows that it can be well used for the study of SARS-CoV entry and inhibition.</AbstractText>
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<PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2006</Year>
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<Day>19</Day>
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<PubMedPubDate PubStatus="medline"><Year>2006</Year>
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<ArticleIdList><ArticleId IdType="pubmed">16916786</ArticleId>
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<ArticleId IdType="doi">10.1080/15216540600820974</ArticleId>
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<affiliations><list><country><li>République populaire de Chine</li>
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<region><li>Hubei</li>
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<settlement><li>Wuhan</li>
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<orgName><li>Université de Wuhan</li>
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<name sortKey="Jiang, Dahe" sort="Jiang, Dahe" uniqKey="Jiang D" first="Dahe" last="Jiang">Dahe Jiang</name>
<name sortKey="Li, Wenxin" sort="Li, Wenxin" uniqKey="Li W" first="Wenxin" last="Li">Wenxin Li</name>
<name sortKey="Liu, Hui" sort="Liu, Hui" uniqKey="Liu H" first="Hui" last="Liu">Hui Liu</name>
<name sortKey="Mao, Xin" sort="Mao, Xin" uniqKey="Mao X" first="Xin" last="Mao">Xin Mao</name>
<name sortKey="Wu, Wenlan" sort="Wu, Wenlan" uniqKey="Wu W" first="Wenlan" last="Wu">Wenlan Wu</name>
<name sortKey="Wu, Yingliang" sort="Wu, Yingliang" uniqKey="Wu Y" first="Yingliang" last="Wu">Yingliang Wu</name>
<name sortKey="Xu, Xiuling" sort="Xu, Xiuling" uniqKey="Xu X" first="Xiuling" last="Xu">Xiuling Xu</name>
<name sortKey="Zhu, Ying" sort="Zhu, Ying" uniqKey="Zhu Y" first="Ying" last="Zhu">Ying Zhu</name>
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<country name="République populaire de Chine"><region name="Hubei"><name sortKey="Sha, Yonggang" sort="Sha, Yonggang" uniqKey="Sha Y" first="Yonggang" last="Sha">Yonggang Sha</name>
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