Establishment of a novel and highly permissive cell line for the efficient replication of cyprinid herpesvirus 2 (CyHV-2)
•A novel cell line from the brain of gibel carp (GiCB) has been established and characterized.•The GiCB cell line can consistently propagate CyHV-2 to high concentrations.•The ultrastructural morphogenesis of CyHV-2 in an established cell line was first observed by TEM.•The CyHV-2 virion produced in...
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| Veröffentlicht in: | Veterinary microbiology 2015-06, Vol.177 (3-4), p.315-325 |
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| creator | Ma, Jie Jiang, Nan LaPatra, Scott E. Jin, Ling Xu, Jin Fan, Yuding Zhou, Yong Zeng, Lingbing |
| description | •A novel cell line from the brain of gibel carp (GiCB) has been established and characterized.•The GiCB cell line can consistently propagate CyHV-2 to high concentrations.•The ultrastructural morphogenesis of CyHV-2 in an established cell line was first observed by TEM.•The CyHV-2 virion produced in GiCB cells is highly pathogenic to gibel carp.
Haematopoietic necrosis of gibel carp (Carassius auratus gibelio) is caused by cyprinid herpesvirus 2 (CyHV-2) and has caused huge economic losses in aquaculture worldwide. Currently the isolation and propagation of CyHV-2 in vitro is very difficult due to the lack of permissive cell lines. Studies on the pathogenesis of CyHV-2 have been hampered because the virus has not been extensively characterized. In this study, a novel cell line from the brain of gibel carp, denoted GiCB, has been established and characterized. Sustainable propagation of CyHV-2 in the GiCB cell line has been confirmed by virus infection and titration, PCR, transmission electron microscopy, immunofluorescence assay and fluorescence in situ hybridization. The GiCB cells showed typical cytopathic effect by day 6 post-infection with CyHV-2 including cell shrinkage, rounding, and cell fusion with cytoplasmic vacuolization. The virus titer reached 107.5±0.37TCID50/ml and has been successfully passaged over 50 times in the GiCB cell line. Electron microscopy analysis revealed the complete replication of CyHV-2 in GiCB cells. CyHV-2-infected GiCB cells reacted strongly with polyclonal antibodies against CyHV-2 and CyHV-2 RNA in cells hybridized specifically with the virus RNA probes. Additionally, an experimental infection demonstrated that CyHV-2 produced in GiCB cells caused 100% mortality in gibel carp. All the results provide solid evidence that the GiCB cell line is highly permissive for the isolation and propagation of CyHV-2. This is a significant advancement that will promote additional research on CyHV-2 infection in fish in the future. |
| doi_str_mv | 10.1016/j.vetmic.2015.04.006 |
| format | Article |
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Haematopoietic necrosis of gibel carp (Carassius auratus gibelio) is caused by cyprinid herpesvirus 2 (CyHV-2) and has caused huge economic losses in aquaculture worldwide. Currently the isolation and propagation of CyHV-2 in vitro is very difficult due to the lack of permissive cell lines. Studies on the pathogenesis of CyHV-2 have been hampered because the virus has not been extensively characterized. In this study, a novel cell line from the brain of gibel carp, denoted GiCB, has been established and characterized. Sustainable propagation of CyHV-2 in the GiCB cell line has been confirmed by virus infection and titration, PCR, transmission electron microscopy, immunofluorescence assay and fluorescence in situ hybridization. The GiCB cells showed typical cytopathic effect by day 6 post-infection with CyHV-2 including cell shrinkage, rounding, and cell fusion with cytoplasmic vacuolization. The virus titer reached 107.5±0.37TCID50/ml and has been successfully passaged over 50 times in the GiCB cell line. Electron microscopy analysis revealed the complete replication of CyHV-2 in GiCB cells. CyHV-2-infected GiCB cells reacted strongly with polyclonal antibodies against CyHV-2 and CyHV-2 RNA in cells hybridized specifically with the virus RNA probes. Additionally, an experimental infection demonstrated that CyHV-2 produced in GiCB cells caused 100% mortality in gibel carp. All the results provide solid evidence that the GiCB cell line is highly permissive for the isolation and propagation of CyHV-2. This is a significant advancement that will promote additional research on CyHV-2 infection in fish in the future.</description><identifier>ISSN: 0378-1135</identifier><identifier>EISSN: 1873-2542</identifier><identifier>DOI: 10.1016/j.vetmic.2015.04.006</identifier><identifier>PMID: 25912023</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Aquaculture ; Brain ; Brain - cytology ; Brain - virology ; Cell line ; Cell Line - cytology ; Cell Line - virology ; Cyprinid herpesvirus 2 (CyHV-2) ; DNA Helicases - genetics ; DNA, Viral - isolation & purification ; Fish Diseases - virology ; Gene Expression Regulation, Viral ; Gibel carp (Carassius auratus gibelio) ; Goldfish - virology ; Herpesviridae - enzymology ; Herpesviridae - genetics ; Herpesviridae - pathogenicity ; Herpesviridae - physiology ; Herpesviridae Infections - veterinary ; Herpesviridae Infections - virology ; In Situ Hybridization, Fluorescence ; Kidney - virology ; Rabbits ; Replication ; Spleen - virology ; Viral Proteins - genetics ; Viral Proteins - metabolism ; Virus Replication</subject><ispartof>Veterinary microbiology, 2015-06, Vol.177 (3-4), p.315-325</ispartof><rights>2015 Elsevier B.V.</rights><rights>Copyright © 2015 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c498t-63e169b9f57e37369d65568f844e4a5b5eca280ea08c41e62c5afe00e6c504033</citedby><cites>FETCH-LOGICAL-c498t-63e169b9f57e37369d65568f844e4a5b5eca280ea08c41e62c5afe00e6c504033</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0378113515001510$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27903,27904,65309</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25912023$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ma, Jie</creatorcontrib><creatorcontrib>Jiang, Nan</creatorcontrib><creatorcontrib>LaPatra, Scott E.</creatorcontrib><creatorcontrib>Jin, Ling</creatorcontrib><creatorcontrib>Xu, Jin</creatorcontrib><creatorcontrib>Fan, Yuding</creatorcontrib><creatorcontrib>Zhou, Yong</creatorcontrib><creatorcontrib>Zeng, Lingbing</creatorcontrib><title>Establishment of a novel and highly permissive cell line for the efficient replication of cyprinid herpesvirus 2 (CyHV-2)</title><title>Veterinary microbiology</title><addtitle>Vet Microbiol</addtitle><description>•A novel cell line from the brain of gibel carp (GiCB) has been established and characterized.•The GiCB cell line can consistently propagate CyHV-2 to high concentrations.•The ultrastructural morphogenesis of CyHV-2 in an established cell line was first observed by TEM.•The CyHV-2 virion produced in GiCB cells is highly pathogenic to gibel carp.
Haematopoietic necrosis of gibel carp (Carassius auratus gibelio) is caused by cyprinid herpesvirus 2 (CyHV-2) and has caused huge economic losses in aquaculture worldwide. Currently the isolation and propagation of CyHV-2 in vitro is very difficult due to the lack of permissive cell lines. Studies on the pathogenesis of CyHV-2 have been hampered because the virus has not been extensively characterized. In this study, a novel cell line from the brain of gibel carp, denoted GiCB, has been established and characterized. Sustainable propagation of CyHV-2 in the GiCB cell line has been confirmed by virus infection and titration, PCR, transmission electron microscopy, immunofluorescence assay and fluorescence in situ hybridization. The GiCB cells showed typical cytopathic effect by day 6 post-infection with CyHV-2 including cell shrinkage, rounding, and cell fusion with cytoplasmic vacuolization. The virus titer reached 107.5±0.37TCID50/ml and has been successfully passaged over 50 times in the GiCB cell line. Electron microscopy analysis revealed the complete replication of CyHV-2 in GiCB cells. CyHV-2-infected GiCB cells reacted strongly with polyclonal antibodies against CyHV-2 and CyHV-2 RNA in cells hybridized specifically with the virus RNA probes. Additionally, an experimental infection demonstrated that CyHV-2 produced in GiCB cells caused 100% mortality in gibel carp. All the results provide solid evidence that the GiCB cell line is highly permissive for the isolation and propagation of CyHV-2. This is a significant advancement that will promote additional research on CyHV-2 infection in fish in the future.</description><subject>Animals</subject><subject>Aquaculture</subject><subject>Brain</subject><subject>Brain - cytology</subject><subject>Brain - virology</subject><subject>Cell line</subject><subject>Cell Line - cytology</subject><subject>Cell Line - virology</subject><subject>Cyprinid herpesvirus 2 (CyHV-2)</subject><subject>DNA Helicases - genetics</subject><subject>DNA, Viral - isolation & purification</subject><subject>Fish Diseases - virology</subject><subject>Gene Expression Regulation, Viral</subject><subject>Gibel carp (Carassius auratus gibelio)</subject><subject>Goldfish - virology</subject><subject>Herpesviridae - enzymology</subject><subject>Herpesviridae - genetics</subject><subject>Herpesviridae - pathogenicity</subject><subject>Herpesviridae - physiology</subject><subject>Herpesviridae Infections - veterinary</subject><subject>Herpesviridae Infections - virology</subject><subject>In Situ Hybridization, Fluorescence</subject><subject>Kidney - virology</subject><subject>Rabbits</subject><subject>Replication</subject><subject>Spleen - virology</subject><subject>Viral Proteins - genetics</subject><subject>Viral Proteins - metabolism</subject><subject>Virus Replication</subject><issn>0378-1135</issn><issn>1873-2542</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kEtv1DAUhS1URKeFf4CQl2WR9Po5zgapGg0tUqVugK3lca4Zj_LCzkTKvyfRFJas7uacc_V9hHxkUDJg-v5UTji20ZccmCpBlgD6DdkwsxUFV5JfkQ2IrSkYE-qa3OR8AgBZaXhHrrmqGAcuNmTe59EdmpiPLXYj7QN1tOsnbKjranqMv47NTAdMbcw5Tkg9Ng1tYoc09ImOR6QYQvRxLSccmujdGPtuHfLzkGIXlxVMA-YppnOmnN7t5qefBf_8nrwNrsn44fXekh9f9993T8Xzy-O33cNz4WVlxkILZLo6VEFtUWyFrmqtlDbBSInSqYNC77gBdGC8ZKi5Vy4gAGqvQIIQt-Tusjuk_vcZ82gXlhXDddifs2XaADPcSL1E5SXqU59zwmAXgtal2TKwq3R7shfpdpVuQdpF-lL79PrhfGix_lf6a3kJfLkEcOGcIiabV2Me65jQj7bu4_8__AE1vZUr</recordid><startdate>20150612</startdate><enddate>20150612</enddate><creator>Ma, Jie</creator><creator>Jiang, Nan</creator><creator>LaPatra, Scott E.</creator><creator>Jin, Ling</creator><creator>Xu, Jin</creator><creator>Fan, Yuding</creator><creator>Zhou, Yong</creator><creator>Zeng, Lingbing</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20150612</creationdate><title>Establishment of a novel and highly permissive cell line for the efficient replication of cyprinid herpesvirus 2 (CyHV-2)</title><author>Ma, Jie ; Jiang, Nan ; LaPatra, Scott E. ; Jin, Ling ; Xu, Jin ; Fan, Yuding ; Zhou, Yong ; Zeng, Lingbing</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c498t-63e169b9f57e37369d65568f844e4a5b5eca280ea08c41e62c5afe00e6c504033</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Animals</topic><topic>Aquaculture</topic><topic>Brain</topic><topic>Brain - cytology</topic><topic>Brain - virology</topic><topic>Cell line</topic><topic>Cell Line - cytology</topic><topic>Cell Line - virology</topic><topic>Cyprinid herpesvirus 2 (CyHV-2)</topic><topic>DNA Helicases - genetics</topic><topic>DNA, Viral - isolation & purification</topic><topic>Fish Diseases - virology</topic><topic>Gene Expression Regulation, Viral</topic><topic>Gibel carp (Carassius auratus gibelio)</topic><topic>Goldfish - virology</topic><topic>Herpesviridae - enzymology</topic><topic>Herpesviridae - genetics</topic><topic>Herpesviridae - pathogenicity</topic><topic>Herpesviridae - physiology</topic><topic>Herpesviridae Infections - veterinary</topic><topic>Herpesviridae Infections - virology</topic><topic>In Situ Hybridization, Fluorescence</topic><topic>Kidney - virology</topic><topic>Rabbits</topic><topic>Replication</topic><topic>Spleen - virology</topic><topic>Viral Proteins - genetics</topic><topic>Viral Proteins - metabolism</topic><topic>Virus Replication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ma, Jie</creatorcontrib><creatorcontrib>Jiang, Nan</creatorcontrib><creatorcontrib>LaPatra, Scott E.</creatorcontrib><creatorcontrib>Jin, Ling</creatorcontrib><creatorcontrib>Xu, Jin</creatorcontrib><creatorcontrib>Fan, Yuding</creatorcontrib><creatorcontrib>Zhou, Yong</creatorcontrib><creatorcontrib>Zeng, Lingbing</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Veterinary microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ma, Jie</au><au>Jiang, Nan</au><au>LaPatra, Scott E.</au><au>Jin, Ling</au><au>Xu, Jin</au><au>Fan, Yuding</au><au>Zhou, Yong</au><au>Zeng, Lingbing</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Establishment of a novel and highly permissive cell line for the efficient replication of cyprinid herpesvirus 2 (CyHV-2)</atitle><jtitle>Veterinary microbiology</jtitle><addtitle>Vet Microbiol</addtitle><date>2015-06-12</date><risdate>2015</risdate><volume>177</volume><issue>3-4</issue><spage>315</spage><epage>325</epage><pages>315-325</pages><issn>0378-1135</issn><eissn>1873-2542</eissn><abstract>•A novel cell line from the brain of gibel carp (GiCB) has been established and characterized.•The GiCB cell line can consistently propagate CyHV-2 to high concentrations.•The ultrastructural morphogenesis of CyHV-2 in an established cell line was first observed by TEM.•The CyHV-2 virion produced in GiCB cells is highly pathogenic to gibel carp.
Haematopoietic necrosis of gibel carp (Carassius auratus gibelio) is caused by cyprinid herpesvirus 2 (CyHV-2) and has caused huge economic losses in aquaculture worldwide. Currently the isolation and propagation of CyHV-2 in vitro is very difficult due to the lack of permissive cell lines. Studies on the pathogenesis of CyHV-2 have been hampered because the virus has not been extensively characterized. In this study, a novel cell line from the brain of gibel carp, denoted GiCB, has been established and characterized. Sustainable propagation of CyHV-2 in the GiCB cell line has been confirmed by virus infection and titration, PCR, transmission electron microscopy, immunofluorescence assay and fluorescence in situ hybridization. The GiCB cells showed typical cytopathic effect by day 6 post-infection with CyHV-2 including cell shrinkage, rounding, and cell fusion with cytoplasmic vacuolization. The virus titer reached 107.5±0.37TCID50/ml and has been successfully passaged over 50 times in the GiCB cell line. Electron microscopy analysis revealed the complete replication of CyHV-2 in GiCB cells. CyHV-2-infected GiCB cells reacted strongly with polyclonal antibodies against CyHV-2 and CyHV-2 RNA in cells hybridized specifically with the virus RNA probes. Additionally, an experimental infection demonstrated that CyHV-2 produced in GiCB cells caused 100% mortality in gibel carp. All the results provide solid evidence that the GiCB cell line is highly permissive for the isolation and propagation of CyHV-2. This is a significant advancement that will promote additional research on CyHV-2 infection in fish in the future.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>25912023</pmid><doi>10.1016/j.vetmic.2015.04.006</doi><tpages>11</tpages></addata></record> |
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| subjects | Animals Aquaculture Brain Brain - cytology Brain - virology Cell line Cell Line - cytology Cell Line - virology Cyprinid herpesvirus 2 (CyHV-2) DNA Helicases - genetics DNA, Viral - isolation & purification Fish Diseases - virology Gene Expression Regulation, Viral Gibel carp (Carassius auratus gibelio) Goldfish - virology Herpesviridae - enzymology Herpesviridae - genetics Herpesviridae - pathogenicity Herpesviridae - physiology Herpesviridae Infections - veterinary Herpesviridae Infections - virology In Situ Hybridization, Fluorescence Kidney - virology Rabbits Replication Spleen - virology Viral Proteins - genetics Viral Proteins - metabolism Virus Replication |
| title | Establishment of a novel and highly permissive cell line for the efficient replication of cyprinid herpesvirus 2 (CyHV-2) |
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