Generation and characterization of potential dengue vaccine candidates based on domain III of the envelope protein and the capsid protein of the four serotypes of dengue virus
Dengue is currently one of the most important arthropod-borne diseases, causing up to 25,000 deaths annually. There is currently no vaccine to prevent dengue virus infection, which needs a tetravalent vaccine approach. In this work, we describe the cloning and expression in Escherichia coli of envel...
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| Veröffentlicht in: | Archives of virology 2014-07, Vol.159 (7), p.1629-1640 |
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| creator | Suzarte, Edith Marcos, Ernesto Gil, Lázaro Valdés, Iris Lazo, Laura Ramos, Yassel Pérez, Yusleidi Falcón, Viviana Romero, Yaremis Guzmán, María G González, Sirenia Kourí, Juan Guillén, Gerardo Hermida, Lisset |
| description | Dengue is currently one of the most important arthropod-borne diseases, causing up to 25,000 deaths annually. There is currently no vaccine to prevent dengue virus infection, which needs a tetravalent vaccine approach. In this work, we describe the cloning and expression in Escherichia coli of envelope domain III-capsid chimeric proteins (DIIIC) of the four dengue serotypes as a tetravalent dengue vaccine candidate that is potentially able to generate humoral and cellular immunity. The recombinant proteins were purified to more than 85 % purity and were recognized by anti-dengue mouse and human sera. Mass spectrometry analysis verified the identity of the proteins and the correct formation of the intracatenary disulfide bond in the domain III region. The chimeric DIIIC proteins were also serotype-specific, and in the presence of oligonucleotides, they formed aggregates that were visible by electron microscopy. These results support the future use of DIIIC recombinant chimeric proteins in preclinical studies in mice for assessing their immunogenicity and efficacy. |
| doi_str_mv | 10.1007/s00705-013-1956-4 |
| format | Article |
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There is currently no vaccine to prevent dengue virus infection, which needs a tetravalent vaccine approach. In this work, we describe the cloning and expression in Escherichia coli of envelope domain III-capsid chimeric proteins (DIIIC) of the four dengue serotypes as a tetravalent dengue vaccine candidate that is potentially able to generate humoral and cellular immunity. The recombinant proteins were purified to more than 85 % purity and were recognized by anti-dengue mouse and human sera. Mass spectrometry analysis verified the identity of the proteins and the correct formation of the intracatenary disulfide bond in the domain III region. The chimeric DIIIC proteins were also serotype-specific, and in the presence of oligonucleotides, they formed aggregates that were visible by electron microscopy. These results support the future use of DIIIC recombinant chimeric proteins in preclinical studies in mice for assessing their immunogenicity and efficacy.</description><identifier>ISSN: 0304-8608</identifier><identifier>EISSN: 1432-8798</identifier><identifier>DOI: 10.1007/s00705-013-1956-4</identifier><identifier>PMID: 24420159</identifier><language>eng</language><publisher>Vienna: Springer-Verlag</publisher><subject>Antibodies ; Antigens ; Antigens, Viral - immunology ; arthropod-borne diseases ; Arthropods ; Biomedical and Life Sciences ; Biomedicine ; Capsid Proteins - genetics ; Capsid Proteins - metabolism ; cell-mediated immunity ; Cloning ; Cloning, Molecular ; coat proteins ; dengue ; Dengue fever ; Dengue Vaccines ; Dengue virus ; Dengue Virus - classification ; Dengue Virus - genetics ; Dengue Virus - immunology ; Dengue Virus - metabolism ; E coli ; electron microscopy ; Escherichia coli ; Gene Expression Regulation, Viral - physiology ; Genetic engineering ; humans ; immune response ; Infections ; Infectious Diseases ; mass spectrometry ; Medical Microbiology ; mice ; oligonucleotides ; Original Article ; Plasmids ; Protein Structure, Tertiary ; Proteins ; recombinant fusion proteins ; Recombinant Proteins - immunology ; serotypes ; Serotyping ; Vaccines ; Viral Envelope Proteins - genetics ; Viral Envelope Proteins - metabolism ; Virology</subject><ispartof>Archives of virology, 2014-07, Vol.159 (7), p.1629-1640</ispartof><rights>Springer-Verlag Wien 2014</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c499t-b9a8ec46f710077692681d3d858f4046763683261cc9097aa9f96d0752b00af53</citedby><cites>FETCH-LOGICAL-c499t-b9a8ec46f710077692681d3d858f4046763683261cc9097aa9f96d0752b00af53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00705-013-1956-4$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00705-013-1956-4$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24420159$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Suzarte, Edith</creatorcontrib><creatorcontrib>Marcos, Ernesto</creatorcontrib><creatorcontrib>Gil, Lázaro</creatorcontrib><creatorcontrib>Valdés, Iris</creatorcontrib><creatorcontrib>Lazo, Laura</creatorcontrib><creatorcontrib>Ramos, Yassel</creatorcontrib><creatorcontrib>Pérez, Yusleidi</creatorcontrib><creatorcontrib>Falcón, Viviana</creatorcontrib><creatorcontrib>Romero, Yaremis</creatorcontrib><creatorcontrib>Guzmán, María G</creatorcontrib><creatorcontrib>González, Sirenia</creatorcontrib><creatorcontrib>Kourí, Juan</creatorcontrib><creatorcontrib>Guillén, Gerardo</creatorcontrib><creatorcontrib>Hermida, Lisset</creatorcontrib><title>Generation and characterization of potential dengue vaccine candidates based on domain III of the envelope protein and the capsid protein of the four serotypes of dengue virus</title><title>Archives of virology</title><addtitle>Arch Virol</addtitle><addtitle>Arch Virol</addtitle><description>Dengue is currently one of the most important arthropod-borne diseases, causing up to 25,000 deaths annually. There is currently no vaccine to prevent dengue virus infection, which needs a tetravalent vaccine approach. In this work, we describe the cloning and expression in Escherichia coli of envelope domain III-capsid chimeric proteins (DIIIC) of the four dengue serotypes as a tetravalent dengue vaccine candidate that is potentially able to generate humoral and cellular immunity. The recombinant proteins were purified to more than 85 % purity and were recognized by anti-dengue mouse and human sera. Mass spectrometry analysis verified the identity of the proteins and the correct formation of the intracatenary disulfide bond in the domain III region. The chimeric DIIIC proteins were also serotype-specific, and in the presence of oligonucleotides, they formed aggregates that were visible by electron microscopy. These results support the future use of DIIIC recombinant chimeric proteins in preclinical studies in mice for assessing their immunogenicity and efficacy.</description><subject>Antibodies</subject><subject>Antigens</subject><subject>Antigens, Viral - immunology</subject><subject>arthropod-borne diseases</subject><subject>Arthropods</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Capsid Proteins - genetics</subject><subject>Capsid Proteins - metabolism</subject><subject>cell-mediated immunity</subject><subject>Cloning</subject><subject>Cloning, Molecular</subject><subject>coat proteins</subject><subject>dengue</subject><subject>Dengue fever</subject><subject>Dengue Vaccines</subject><subject>Dengue virus</subject><subject>Dengue Virus - classification</subject><subject>Dengue Virus - genetics</subject><subject>Dengue Virus - immunology</subject><subject>Dengue Virus - metabolism</subject><subject>E coli</subject><subject>electron microscopy</subject><subject>Escherichia coli</subject><subject>Gene Expression Regulation, Viral - physiology</subject><subject>Genetic engineering</subject><subject>humans</subject><subject>immune response</subject><subject>Infections</subject><subject>Infectious Diseases</subject><subject>mass spectrometry</subject><subject>Medical Microbiology</subject><subject>mice</subject><subject>oligonucleotides</subject><subject>Original Article</subject><subject>Plasmids</subject><subject>Protein Structure, Tertiary</subject><subject>Proteins</subject><subject>recombinant fusion proteins</subject><subject>Recombinant Proteins - immunology</subject><subject>serotypes</subject><subject>Serotyping</subject><subject>Vaccines</subject><subject>Viral Envelope Proteins - genetics</subject><subject>Viral Envelope Proteins - metabolism</subject><subject>Virology</subject><issn>0304-8608</issn><issn>1432-8798</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNkc1u1DAUhSMEokPhAdiAJTbdBPz_s0QVlJEqsYCuLY99M3WVcYKdVCovxSviNNMKsUBsbOnc75zr69s0rwl-TzBWH0o9sGgxYS0xQrb8SbMhnNFWK6OfNhvMMG-1xPqkeVHKDcZVYOJ5c0I5p5gIs2l-XUCC7KY4JORSQP7aZecnyPHnKg4dGocJ0hRdjwKk_Qzo1nkfEyBfHTG4CQrauQIBVT4MBxcT2m63i3W6BgTpFvphBDTmGhTXPkvBu7HE8Cgf8W6YMypQxbuxBlf1oWvMc3nZPOtcX-DV8T5trj5_-n7-pb38erE9_3jZem7M1O6M0-C57NTyUUoaKjUJLGihO465VJJJzagk3htslHOmMzJgJegOY9cJdtqcrbn1dT9mKJM9xOKh712CYS6WCMENp4qp_0A5YcpQpSv67i_0pk6b6iALhQUl8j6QrJTPQykZOjvmeHD5zhJsl3nsunhbF2-XxVtePW-OyfPuAOHR8bDpCtAVKLWU9pD_aP2P1LerqXODdfsci736VvM4rpBQkrPfAkDCfQ</recordid><startdate>20140701</startdate><enddate>20140701</enddate><creator>Suzarte, Edith</creator><creator>Marcos, Ernesto</creator><creator>Gil, Lázaro</creator><creator>Valdés, Iris</creator><creator>Lazo, Laura</creator><creator>Ramos, Yassel</creator><creator>Pérez, Yusleidi</creator><creator>Falcón, Viviana</creator><creator>Romero, Yaremis</creator><creator>Guzmán, María G</creator><creator>González, Sirenia</creator><creator>Kourí, Juan</creator><creator>Guillén, Gerardo</creator><creator>Hermida, Lisset</creator><general>Springer-Verlag</general><general>Springer Vienna</general><general>Springer Nature B.V</general><scope>FBQ</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>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>RC3</scope><scope>7X8</scope><scope>7T2</scope><scope>7U2</scope><scope>C1K</scope><scope>F1W</scope><scope>H95</scope><scope>H97</scope><scope>L.G</scope></search><sort><creationdate>20140701</creationdate><title>Generation and characterization of potential dengue vaccine candidates based on domain III of the envelope protein and the capsid protein of the four serotypes of dengue virus</title><author>Suzarte, Edith ; Marcos, Ernesto ; Gil, Lázaro ; Valdés, Iris ; Lazo, Laura ; Ramos, Yassel ; Pérez, Yusleidi ; Falcón, Viviana ; Romero, Yaremis ; Guzmán, María G ; González, Sirenia ; Kourí, Juan ; Guillén, Gerardo ; Hermida, Lisset</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c499t-b9a8ec46f710077692681d3d858f4046763683261cc9097aa9f96d0752b00af53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Antibodies</topic><topic>Antigens</topic><topic>Antigens, Viral - 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| ispartof | Archives of virology, 2014-07, Vol.159 (7), p.1629-1640 |
| issn | 0304-8608 1432-8798 |
| language | eng |
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| subjects | Antibodies Antigens Antigens, Viral - immunology arthropod-borne diseases Arthropods Biomedical and Life Sciences Biomedicine Capsid Proteins - genetics Capsid Proteins - metabolism cell-mediated immunity Cloning Cloning, Molecular coat proteins dengue Dengue fever Dengue Vaccines Dengue virus Dengue Virus - classification Dengue Virus - genetics Dengue Virus - immunology Dengue Virus - metabolism E coli electron microscopy Escherichia coli Gene Expression Regulation, Viral - physiology Genetic engineering humans immune response Infections Infectious Diseases mass spectrometry Medical Microbiology mice oligonucleotides Original Article Plasmids Protein Structure, Tertiary Proteins recombinant fusion proteins Recombinant Proteins - immunology serotypes Serotyping Vaccines Viral Envelope Proteins - genetics Viral Envelope Proteins - metabolism Virology |
| title | Generation and characterization of potential dengue vaccine candidates based on domain III of the envelope protein and the capsid protein of the four serotypes of dengue virus |
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