Establishment of Vero E6 cell clones persistently infected with severe acute respiratory syndrome coronavirus.
Identifieur interne : 001249 ( Ncbi/Merge ); 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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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></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><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><term>Régulation négative</term><term>Technique de Western</term></keywords></textClass></profileDesc></teiHeader><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></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16269264</PMID><DateCompleted><Year>2006</Year><Month>03</Month><Day>07</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>15</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">1286-4579</ISSN><JournalIssue CitedMedium="Print"><Volume>7</Volume><Issue>15</Issue><PubDate><Year>2005</Year><Month>Dec</Month></PubDate></JournalIssue><Title>Microbes and infection</Title><ISOAbbreviation>Microbes Infect.</ISOAbbreviation></Journal><ArticleTitle>Establishment of Vero E6 cell clones persistently infected with severe acute respiratory syndrome coronavirus.</ArticleTitle><Pagination><MedlinePgn>1530-40</MedlinePgn></Pagination><Abstract><AbstractText>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.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Yamate</LastName><ForeName>Masanobu</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yamashita</LastName><ForeName>Makiko</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Goto</LastName><ForeName>Toshiyuki</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Tsuji</LastName><ForeName>Shoutaro</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Yong-Gang</ForeName><Initials>YG</Initials></Author><Author ValidYN="Y"><LastName>Warachit</LastName><ForeName>Jiranan</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Yunoki</LastName><ForeName>Mikihiro</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Ikuta</LastName><ForeName>Kazuyoshi</ForeName><Initials>K</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2005</Year><Month>07</Month><Day>06</Day></ArticleDate></Article><MedlineJournalInfo><Country>France</Country><MedlineTA>Microbes Infect</MedlineTA><NlmUniqueID>100883508</NlmUniqueID><ISSNLinking>1286-4579</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000954">Antigens, Surface</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D008562">Membrane Glycoproteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D019590">Nucleocapsid Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D012367">RNA, Viral</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D064370">Spike Glycoprotein, Coronavirus</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D014759">Viral Envelope Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C578557">spike glycoprotein, SARS-CoV</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.4.-</RegistryNumber><NameOfSubstance UI="D002268">Carboxypeptidases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.4.15.1</RegistryNumber><NameOfSubstance UI="D007703">Peptidyl-Dipeptidase A</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.4.17.-</RegistryNumber><NameOfSubstance UI="C413524">angiotensin converting enzyme 2</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000954" MajorTopicYN="N">Antigens, Surface</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015153" MajorTopicYN="N">Blotting, Western</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002268" MajorTopicYN="N">Carboxypeptidases</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D002522" MajorTopicYN="N">Chlorocebus aethiops</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D003588" MajorTopicYN="N">Cytopathogenic Effect, Viral</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015536" MajorTopicYN="N">Down-Regulation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005434" MajorTopicYN="N">Flow Cytometry</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008562" MajorTopicYN="N">Membrane Glycoproteins</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018613" MajorTopicYN="N">Microscopy, Confocal</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008854" MajorTopicYN="N">Microscopy, Electron</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008856" MajorTopicYN="N">Microscopy, Fluorescence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019590" MajorTopicYN="N">Nucleocapsid Proteins</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D007703" MajorTopicYN="N">Peptidyl-Dipeptidase A</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012367" MajorTopicYN="N">RNA, Viral</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D045473" MajorTopicYN="N">SARS Virus</DescriptorName><QualifierName UI="Q000254" MajorTopicYN="Y">growth & development</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D064370" MajorTopicYN="N">Spike Glycoprotein, Coronavirus</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014709" MajorTopicYN="N">Vero Cells</DescriptorName><QualifierName UI="Q000821" MajorTopicYN="Y">virology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014759" MajorTopicYN="N">Viral Envelope Proteins</DescriptorName><QualifierName UI="Q000032" 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first="Toshiyuki" last="Goto">Toshiyuki Goto</name><name sortKey="Ikuta, Kazuyoshi" sort="Ikuta, Kazuyoshi" uniqKey="Ikuta K" first="Kazuyoshi" last="Ikuta">Kazuyoshi Ikuta</name><name sortKey="Li, Yong Gang" sort="Li, Yong Gang" uniqKey="Li Y" first="Yong-Gang" last="Li">Yong-Gang Li</name><name sortKey="Tsuji, Shoutaro" sort="Tsuji, Shoutaro" uniqKey="Tsuji S" first="Shoutaro" last="Tsuji">Shoutaro Tsuji</name><name sortKey="Warachit, Jiranan" sort="Warachit, Jiranan" uniqKey="Warachit J" first="Jiranan" last="Warachit">Jiranan Warachit</name><name sortKey="Yamashita, Makiko" sort="Yamashita, Makiko" uniqKey="Yamashita M" first="Makiko" last="Yamashita">Makiko Yamashita</name><name sortKey="Yunoki, Mikihiro" sort="Yunoki, Mikihiro" uniqKey="Yunoki M" first="Mikihiro" last="Yunoki">Mikihiro Yunoki</name></noCountry><country name="Japon"><noRegion><name sortKey="Yamate, Masanobu" sort="Yamate, Masanobu" uniqKey="Yamate M" first="Masanobu" last="Yamate">Masanobu 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