Rabies virus-like particles expressed in HEK293 cells
Abstract Rabies is an infectious viral disease with a mortality rate close to 100%. Currently, there is a need to generate cheaper and more immunogenic vaccines for the effective prevention of rabies, mostly in developing countries. Virus-like particles have been widely used in viral vaccine product...
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| Veröffentlicht in: | Vaccine 2014-05, Vol.32 (24), p.2799-2804 |
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| creator | Fontana, Diego Kratje, Ricardo Etcheverrigaray, Marina Prieto, Claudio |
| description | Abstract Rabies is an infectious viral disease with a mortality rate close to 100%. Currently, there is a need to generate cheaper and more immunogenic vaccines for the effective prevention of rabies, mostly in developing countries. Virus-like particles have been widely used in viral vaccine production due to their high immunogenicity and safety during the production process. Rabies virus glycoprotein is the major antigen to trigger a protective immune response and the only protein capable of generating virus neutralizing antibodies. In this study we describe the development of a recombinant stable cell line for the production of rabies virus-like particles (RV-VLPs) expressing the rabies virus glycoprotein by lentivirus-based transduction of HEK293 cells. Protein expression was analyzed by flow cytometry, fluorescence microscopy, western blot and ELISA. Particles were purified from culture supernatant and their size and morphology were studied. Furthermore, mice were immunized with RV-VLPs, formulated with adjuvant, and these particles were able to produce a specific antibody response, demonstrating that these virus-like particles present a promising rabies vaccine candidate. |
| doi_str_mv | 10.1016/j.vaccine.2014.02.031 |
| format | Article |
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Currently, there is a need to generate cheaper and more immunogenic vaccines for the effective prevention of rabies, mostly in developing countries. Virus-like particles have been widely used in viral vaccine production due to their high immunogenicity and safety during the production process. Rabies virus glycoprotein is the major antigen to trigger a protective immune response and the only protein capable of generating virus neutralizing antibodies. In this study we describe the development of a recombinant stable cell line for the production of rabies virus-like particles (RV-VLPs) expressing the rabies virus glycoprotein by lentivirus-based transduction of HEK293 cells. Protein expression was analyzed by flow cytometry, fluorescence microscopy, western blot and ELISA. Particles were purified from culture supernatant and their size and morphology were studied. Furthermore, mice were immunized with RV-VLPs, formulated with adjuvant, and these particles were able to produce a specific antibody response, demonstrating that these virus-like particles present a promising rabies vaccine candidate.</description><identifier>ISSN: 0264-410X</identifier><identifier>EISSN: 1873-2518</identifier><identifier>DOI: 10.1016/j.vaccine.2014.02.031</identifier><identifier>PMID: 24631077</identifier><identifier>CODEN: VACCDE</identifier><language>eng</language><publisher>Kidlington: Elsevier Ltd</publisher><subject>Allergy and Immunology ; Animals ; Antibodies, Viral - blood ; Antigens, Viral - immunology ; Applied microbiology ; Biological and medical sciences ; Cloning ; Developing countries ; Female ; Fluorescence microscopy ; Fundamental and applied biological sciences. Psychology ; Genetic Vectors ; Glycoproteins - immunology ; HEK293 Cells ; Humans ; Immune response ; Immunity, Humoral ; Immunization ; Immunogenicity ; Industrialized nations ; LDCs ; Mice ; Mice, Inbred BALB C ; Microbiology ; Miscellaneous ; Plasmids ; Proteins ; Rabies ; Rabies Vaccines - immunology ; Rabies virus ; Rabies virus - growth & development ; RNA polymerase ; Transduction, Genetic ; Vaccine ; Vaccines ; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects) ; Vaccines, Virus-Like Particle - immunology ; Viral diseases ; Viral Envelope Proteins - immunology ; Virology ; Virus Cultivation ; Virus-like particles ; Viruses</subject><ispartof>Vaccine, 2014-05, Vol.32 (24), p.2799-2804</ispartof><rights>Elsevier Ltd</rights><rights>2014 Elsevier Ltd</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2014 Elsevier Ltd. All rights reserved.</rights><rights>Copyright Elsevier Limited May 19, 2014</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c558t-ba455d91d9679e28b6b1214b1a082a8ccc81349e503ee0cde6e8866a59865c603</citedby><cites>FETCH-LOGICAL-c558t-ba455d91d9679e28b6b1214b1a082a8ccc81349e503ee0cde6e8866a59865c603</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0264410X14002047$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>309,310,314,776,780,785,786,3536,23910,23911,25119,27903,27904,65309</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=28469441$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24631077$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fontana, Diego</creatorcontrib><creatorcontrib>Kratje, Ricardo</creatorcontrib><creatorcontrib>Etcheverrigaray, Marina</creatorcontrib><creatorcontrib>Prieto, Claudio</creatorcontrib><title>Rabies virus-like particles expressed in HEK293 cells</title><title>Vaccine</title><addtitle>Vaccine</addtitle><description>Abstract Rabies is an infectious viral disease with a mortality rate close to 100%. Currently, there is a need to generate cheaper and more immunogenic vaccines for the effective prevention of rabies, mostly in developing countries. Virus-like particles have been widely used in viral vaccine production due to their high immunogenicity and safety during the production process. Rabies virus glycoprotein is the major antigen to trigger a protective immune response and the only protein capable of generating virus neutralizing antibodies. In this study we describe the development of a recombinant stable cell line for the production of rabies virus-like particles (RV-VLPs) expressing the rabies virus glycoprotein by lentivirus-based transduction of HEK293 cells. Protein expression was analyzed by flow cytometry, fluorescence microscopy, western blot and ELISA. Particles were purified from culture supernatant and their size and morphology were studied. Furthermore, mice were immunized with RV-VLPs, formulated with adjuvant, and these particles were able to produce a specific antibody response, demonstrating that these virus-like particles present a promising rabies vaccine candidate.</description><subject>Allergy and Immunology</subject><subject>Animals</subject><subject>Antibodies, Viral - blood</subject><subject>Antigens, Viral - immunology</subject><subject>Applied microbiology</subject><subject>Biological and medical sciences</subject><subject>Cloning</subject><subject>Developing countries</subject><subject>Female</subject><subject>Fluorescence microscopy</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetic Vectors</subject><subject>Glycoproteins - immunology</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>Immune response</subject><subject>Immunity, Humoral</subject><subject>Immunization</subject><subject>Immunogenicity</subject><subject>Industrialized nations</subject><subject>LDCs</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Rabies</subject><subject>Rabies Vaccines - immunology</subject><subject>Rabies virus</subject><subject>Rabies virus - growth & development</subject><subject>RNA polymerase</subject><subject>Transduction, Genetic</subject><subject>Vaccine</subject><subject>Vaccines</subject><subject>Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects)</subject><subject>Vaccines, Virus-Like Particle - immunology</subject><subject>Viral diseases</subject><subject>Viral Envelope Proteins - immunology</subject><subject>Virology</subject><subject>Virus Cultivation</subject><subject>Virus-like particles</subject><subject>Viruses</subject><issn>0264-410X</issn><issn>1873-2518</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>BENPR</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqNkl1rFDEUhoNY7Lb6E5QBEXoz4zmZJJPcWKRUWywIfoB3IZM5C9nOzqzJztL--2bY1UJv9CoQnvc9H-9h7DVChYDq_araOe_DQBUHFBXwCmp8xhaom7rkEvVztgCuRCkQfh2zk5RWACBrNC_YMReqRmiaBZPfXBsoFbsQp1T24ZaKjYvb4Pv8SXebSClRV4ShuLr8wk1deOr79JIdLV2f6NXhPWU_P13-uLgqb75-vr74eFN6KfW2bJ2QsjPYGdUY4rpVLXIULTrQ3GnvvcZaGJJQE4HvSJHWSjlptJJeQX3Kzva-mzj-niht7TqkuQM30Dgli5ILAdJw8T9orsWh0Rl9-wRdjVMc8iAzBdgoNHNtuad8HFOKtLSbGNYu3lsEO0dgV_YQgZ0jsMBtjiDr3hzcp3ZN3V_Vn51n4N0BcMm7fhnd4EN65LRQRojZ6HzPUd7wLlC0yQcaPHUhkt_abgz_bOXDEwffhyHkord0T-lxapuywH6f72U-FxQAHERTPwCs6LfL</recordid><startdate>20140519</startdate><enddate>20140519</enddate><creator>Fontana, Diego</creator><creator>Kratje, Ricardo</creator><creator>Etcheverrigaray, Marina</creator><creator>Prieto, Claudio</creator><general>Elsevier Ltd</general><general>Elsevier</general><general>Elsevier Limited</general><scope>IQODW</scope><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>3V.</scope><scope>7QL</scope><scope>7RV</scope><scope>7T2</scope><scope>7T5</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88C</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9-</scope><scope>K9.</scope><scope>KB0</scope><scope>LK8</scope><scope>M0R</scope><scope>M0S</scope><scope>M0T</scope><scope>M1P</scope><scope>M2O</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope></search><sort><creationdate>20140519</creationdate><title>Rabies virus-like particles expressed in HEK293 cells</title><author>Fontana, Diego ; Kratje, Ricardo ; Etcheverrigaray, Marina ; Prieto, Claudio</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c558t-ba455d91d9679e28b6b1214b1a082a8ccc81349e503ee0cde6e8866a59865c603</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Allergy and Immunology</topic><topic>Animals</topic><topic>Antibodies, Viral - blood</topic><topic>Antigens, Viral - immunology</topic><topic>Applied microbiology</topic><topic>Biological and medical sciences</topic><topic>Cloning</topic><topic>Developing countries</topic><topic>Female</topic><topic>Fluorescence microscopy</topic><topic>Fundamental and applied biological sciences. 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Academic</collection><jtitle>Vaccine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Fontana, Diego</au><au>Kratje, Ricardo</au><au>Etcheverrigaray, Marina</au><au>Prieto, Claudio</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rabies virus-like particles expressed in HEK293 cells</atitle><jtitle>Vaccine</jtitle><addtitle>Vaccine</addtitle><date>2014-05-19</date><risdate>2014</risdate><volume>32</volume><issue>24</issue><spage>2799</spage><epage>2804</epage><pages>2799-2804</pages><issn>0264-410X</issn><eissn>1873-2518</eissn><coden>VACCDE</coden><abstract>Abstract Rabies is an infectious viral disease with a mortality rate close to 100%. Currently, there is a need to generate cheaper and more immunogenic vaccines for the effective prevention of rabies, mostly in developing countries. Virus-like particles have been widely used in viral vaccine production due to their high immunogenicity and safety during the production process. Rabies virus glycoprotein is the major antigen to trigger a protective immune response and the only protein capable of generating virus neutralizing antibodies. In this study we describe the development of a recombinant stable cell line for the production of rabies virus-like particles (RV-VLPs) expressing the rabies virus glycoprotein by lentivirus-based transduction of HEK293 cells. Protein expression was analyzed by flow cytometry, fluorescence microscopy, western blot and ELISA. Particles were purified from culture supernatant and their size and morphology were studied. Furthermore, mice were immunized with RV-VLPs, formulated with adjuvant, and these particles were able to produce a specific antibody response, demonstrating that these virus-like particles present a promising rabies vaccine candidate.</abstract><cop>Kidlington</cop><pub>Elsevier Ltd</pub><pmid>24631077</pmid><doi>10.1016/j.vaccine.2014.02.031</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Allergy and Immunology Animals Antibodies, Viral - blood Antigens, Viral - immunology Applied microbiology Biological and medical sciences Cloning Developing countries Female Fluorescence microscopy Fundamental and applied biological sciences. Psychology Genetic Vectors Glycoproteins - immunology HEK293 Cells Humans Immune response Immunity, Humoral Immunization Immunogenicity Industrialized nations LDCs Mice Mice, Inbred BALB C Microbiology Miscellaneous Plasmids Proteins Rabies Rabies Vaccines - immunology Rabies virus Rabies virus - growth & development RNA polymerase Transduction, Genetic Vaccine Vaccines Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects) Vaccines, Virus-Like Particle - immunology Viral diseases Viral Envelope Proteins - immunology Virology Virus Cultivation Virus-like particles Viruses |
| title | Rabies virus-like particles expressed in HEK293 cells |
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