Establishment of Vero E6 cell clones persistently infected with severe acute respiratory syndrome coronavirus.
Identifieur interne : 001249 ( Ncbi/Curation ); précédent : 001248; suivant : 001250Establishment of Vero E6 cell clones persistently infected with severe acute respiratory syndrome coronavirus.
Auteurs : Masanobu Yamate [Japon] ; Makiko Yamashita ; Toshiyuki Goto ; Shoutaro Tsuji ; Yong-Gang Li ; Jiranan Warachit ; Mikihiro Yunoki ; Kazuyoshi IkutaSource :
- Microbes and infection [ 1286-4579 ] ; 2005.
Descripteurs français
- KwdFr :
- ARN viral (analyse), Animaux, Antigènes de surface (analyse), Carboxypeptidases (analyse), Cellules Vero (virologie), Cytométrie en flux, Effet cytopathogène viral, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (analyse), Microscopie confocale, Microscopie de fluorescence, Microscopie électronique, Peptidyl-Dipeptidase A, Protéines de l'enveloppe virale (analyse), Protéines nucléocapside (analyse), Régulation négative, Technique de Western, Virus du SRAS (croissance et développement).
- MESH :
- analyse : ARN viral, Antigènes de surface, Carboxypeptidases, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines nucléocapside.
- croissance et développement : Virus du SRAS.
- virologie : Cellules Vero.
- Animaux, Cytométrie en flux, Effet cytopathogène viral, Glycoprotéine de spicule des coronavirus, Microscopie confocale, Microscopie de fluorescence, Microscopie électronique, Peptidyl-Dipeptidase A, Régulation négative, Technique de Western.
English descriptors
- KwdEn :
- Animals, Antigens, Surface (analysis), Blotting, Western, Carboxypeptidases (analysis), Chlorocebus aethiops, Cytopathogenic Effect, Viral, Down-Regulation, Flow Cytometry, Membrane Glycoproteins (analysis), Microscopy, Confocal, Microscopy, Electron, Microscopy, Fluorescence, Nucleocapsid Proteins (analysis), Peptidyl-Dipeptidase A, RNA, Viral (analysis), SARS Virus (growth & development), Spike Glycoprotein, Coronavirus, Vero Cells (virology), Viral Envelope Proteins (analysis).
- MESH :
- chemical , analysis : Antigens, Surface, Carboxypeptidases, Membrane Glycoproteins, Nucleocapsid Proteins, RNA, Viral, Viral Envelope Proteins.
- growth & development : SARS Virus.
- virology : Vero Cells.
- Animals, Blotting, Western, Chlorocebus aethiops, Cytopathogenic Effect, Viral, Down-Regulation, Flow Cytometry, Microscopy, Confocal, Microscopy, Electron, Microscopy, Fluorescence, Peptidyl-Dipeptidase A, Spike Glycoprotein, Coronavirus.
Abstract
Little information is available on persistent infection of severe acute respiratory syndrome (SARS) coronavirus (CoV). In this study, we established persistent infection of SARS-CoV in the Vero E6 cell line. Acute infection of Vero E6 with SARS-CoV produced a lytic infection with characteristic rounding cytopathic effects (CPE) and the production of a large number of infectious particles in the culture fluid within 3 days post-infection. Upon subsequent culturing of the remaining adherent cells, the cells gradually proliferated and recovered normal morphology similar to that of the parental cells, and continued to produce large numbers of infectious viral particles during the observation period of 5 months. Among a total of 87 cell clones obtained from the persistently infected Vero E6, only four cell clones (named #13, #18, #21, and #34) were positive for viral RNA. Clones #13, #18, and #34 shifted to viral RNA-negative during subsequent cultures, while #21 continuously produced infectious particles at a high rate. The SARS-CoV receptor, angiotensin-converting enzyme 2, was almost completely down regulated from the cell surface of persistently infected cells. Western blot analysis as well as electron microscopy indicated that the ratios of spike to nucleocapsid protein in clone #21 as well as its parental persistently infected cells were lower than that in the cells in the acute phase of infection. These Vero E6 cells persistently infected with SARS-CoV may be useful for clarifying the mechanism of the persistent infection and also for elucidating the possible pathophysiologic significance of such long-term maintenance of this virus.
DOI: 10.1016/j.micinf.2005.05.013
PubMed: 16269264
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pubmed:16269264Le document en format XML
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<author><name sortKey="Yamashita, Makiko" sort="Yamashita, Makiko" uniqKey="Yamashita M" first="Makiko" last="Yamashita">Makiko Yamashita</name>
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<author><name sortKey="Li, Yong Gang" sort="Li, Yong Gang" uniqKey="Li Y" first="Yong-Gang" last="Li">Yong-Gang Li</name>
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<author><name sortKey="Yunoki, Mikihiro" sort="Yunoki, Mikihiro" uniqKey="Yunoki M" first="Mikihiro" last="Yunoki">Mikihiro Yunoki</name>
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<term>Antigens, Surface (analysis)</term>
<term>Blotting, Western</term>
<term>Carboxypeptidases (analysis)</term>
<term>Chlorocebus aethiops</term>
<term>Cytopathogenic Effect, Viral</term>
<term>Down-Regulation</term>
<term>Flow Cytometry</term>
<term>Membrane Glycoproteins (analysis)</term>
<term>Microscopy, Confocal</term>
<term>Microscopy, Electron</term>
<term>Microscopy, Fluorescence</term>
<term>Nucleocapsid Proteins (analysis)</term>
<term>Peptidyl-Dipeptidase A</term>
<term>RNA, Viral (analysis)</term>
<term>SARS Virus (growth & development)</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Vero Cells (virology)</term>
<term>Viral Envelope Proteins (analysis)</term>
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<term>Animaux</term>
<term>Antigènes de surface (analyse)</term>
<term>Carboxypeptidases (analyse)</term>
<term>Cellules Vero (virologie)</term>
<term>Cytométrie en flux</term>
<term>Effet cytopathogène viral</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires (analyse)</term>
<term>Microscopie confocale</term>
<term>Microscopie de fluorescence</term>
<term>Microscopie électronique</term>
<term>Peptidyl-Dipeptidase A</term>
<term>Protéines de l'enveloppe virale (analyse)</term>
<term>Protéines nucléocapside (analyse)</term>
<term>Régulation négative</term>
<term>Technique de Western</term>
<term>Virus du SRAS (croissance et développement)</term>
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<term>Carboxypeptidases</term>
<term>Membrane Glycoproteins</term>
<term>Nucleocapsid Proteins</term>
<term>RNA, Viral</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ARN viral</term>
<term>Antigènes de surface</term>
<term>Carboxypeptidases</term>
<term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines nucléocapside</term>
</keywords>
<keywords scheme="MESH" qualifier="croissance et développement" xml:lang="fr"><term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Cellules Vero</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Vero Cells</term>
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<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Blotting, Western</term>
<term>Chlorocebus aethiops</term>
<term>Cytopathogenic Effect, Viral</term>
<term>Down-Regulation</term>
<term>Flow Cytometry</term>
<term>Microscopy, Confocal</term>
<term>Microscopy, Electron</term>
<term>Microscopy, Fluorescence</term>
<term>Peptidyl-Dipeptidase A</term>
<term>Spike Glycoprotein, Coronavirus</term>
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<term>Cytométrie en flux</term>
<term>Effet cytopathogène viral</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Microscopie confocale</term>
<term>Microscopie de fluorescence</term>
<term>Microscopie électronique</term>
<term>Peptidyl-Dipeptidase A</term>
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<front><div type="abstract" xml:lang="en">Little information is available on persistent infection of severe acute respiratory syndrome (SARS) coronavirus (CoV). In this study, we established persistent infection of SARS-CoV in the Vero E6 cell line. Acute infection of Vero E6 with SARS-CoV produced a lytic infection with characteristic rounding cytopathic effects (CPE) and the production of a large number of infectious particles in the culture fluid within 3 days post-infection. Upon subsequent culturing of the remaining adherent cells, the cells gradually proliferated and recovered normal morphology similar to that of the parental cells, and continued to produce large numbers of infectious viral particles during the observation period of 5 months. Among a total of 87 cell clones obtained from the persistently infected Vero E6, only four cell clones (named #13, #18, #21, and #34) were positive for viral RNA. Clones #13, #18, and #34 shifted to viral RNA-negative during subsequent cultures, while #21 continuously produced infectious particles at a high rate. The SARS-CoV receptor, angiotensin-converting enzyme 2, was almost completely down regulated from the cell surface of persistently infected cells. Western blot analysis as well as electron microscopy indicated that the ratios of spike to nucleocapsid protein in clone #21 as well as its parental persistently infected cells were lower than that in the cells in the acute phase of infection. These Vero E6 cells persistently infected with SARS-CoV may be useful for clarifying the mechanism of the persistent infection and also for elucidating the possible pathophysiologic significance of such long-term maintenance of this virus.</div>
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