A suicidal DNA vaccine expressing the fusion protein of peste des petits ruminants virus induces both humoral and cell-mediated immune responses in mice

•A suicidal DNA vaccine against peste des petits ruminants virus (PPRV) was developed.•The suicidal DNA vaccine could express Fusion protein in transfected BHK-21 cells.•Specific humoral and cell-mediated immune responses were induced in immunized mice.•The results provide a new strategy to the deve...

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Veröffentlicht in:Journal of virological methods 2015-12, Vol.225, p.35-40
Hauptverfasser: Wang, Yong, Yue, Xiaolin, Jin, Hongyan, Liu, Guangqing, Pan, Ling, Wang, Guijun, Guo, Hao, Li, Gang, Li, Yongdong
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container_title Journal of virological methods
container_volume 225
creator Wang, Yong
Yue, Xiaolin
Jin, Hongyan
Liu, Guangqing
Pan, Ling
Wang, Guijun
Guo, Hao
Li, Gang
Li, Yongdong
description •A suicidal DNA vaccine against peste des petits ruminants virus (PPRV) was developed.•The suicidal DNA vaccine could express Fusion protein in transfected BHK-21 cells.•Specific humoral and cell-mediated immune responses were induced in immunized mice.•The results provide a new strategy to the development of vaccines against PPRV. Peste des petits ruminants (PPR), a highly contagious disease induced by PPR virus (PPRV), affects sheep and goats. PPRV fusion (F) protein is important for the induction of immune responses against PPRV. We constructed a Semliki Forest virus (SFV) replicon-vectored DNA vaccine (“suicidal DNA vaccine”) and evaluated its immunogenicity in BALB/c mice. The F gene of PPRV was cloned and inserted into the SFV replicon-based vector pSCA1. The antigenicity of the resultant plasmid pSCA1/F was identified by indirect immunofluorescence and western blotting. BALB/c mice were then intramuscularly injected with pSCA1/F three times at 14-d intervals. Specific antibodies and virus-neutralizing antibodies against PPRV were quantified by indirect ELISA and microneutralization tests, respectively. Cell-mediated immune responses were examined by cytokine and lymphocyte proliferation assays. The pSCA1/F expressed F protein in vitro and induced specific and neutralizing antibody production, and lymphocyte proliferation in mice. Mice vaccinated with pSCA1/F had increased IL-2 and IL-10 levels after 24-h post first immunization. IFN-γ and TNF-α levels increased from that time point and gradually decreased thereafter. Thus, the Semliki Forest virus replicon-vectored DNA vaccine expressing the F protein of PPRV induced both humoral and cell-mediated immune responses in mice. This could be considered as a novel strategy for vaccine development against PPR.
doi_str_mv 10.1016/j.jviromet.2015.09.001
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Peste des petits ruminants (PPR), a highly contagious disease induced by PPR virus (PPRV), affects sheep and goats. PPRV fusion (F) protein is important for the induction of immune responses against PPRV. We constructed a Semliki Forest virus (SFV) replicon-vectored DNA vaccine (“suicidal DNA vaccine”) and evaluated its immunogenicity in BALB/c mice. The F gene of PPRV was cloned and inserted into the SFV replicon-based vector pSCA1. The antigenicity of the resultant plasmid pSCA1/F was identified by indirect immunofluorescence and western blotting. BALB/c mice were then intramuscularly injected with pSCA1/F three times at 14-d intervals. Specific antibodies and virus-neutralizing antibodies against PPRV were quantified by indirect ELISA and microneutralization tests, respectively. Cell-mediated immune responses were examined by cytokine and lymphocyte proliferation assays. The pSCA1/F expressed F protein in vitro and induced specific and neutralizing antibody production, and lymphocyte proliferation in mice. Mice vaccinated with pSCA1/F had increased IL-2 and IL-10 levels after 24-h post first immunization. IFN-γ and TNF-α levels increased from that time point and gradually decreased thereafter. Thus, the Semliki Forest virus replicon-vectored DNA vaccine expressing the F protein of PPRV induced both humoral and cell-mediated immune responses in mice. This could be considered as a novel strategy for vaccine development against PPR.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/j.jviromet.2015.09.001</identifier><identifier>PMID: 26343487</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Antibodies, Neutralizing - blood ; Antibodies, Viral - blood ; Cell Proliferation ; Cloning, Molecular ; Cytokines - secretion ; Enzyme-Linked Immunosorbent Assay ; Fusion protein ; Immune response ; Immunization Schedule ; Injections, Intramuscular ; Leukocytes, Mononuclear - immunology ; Mice, Inbred BALB C ; Neutralization Tests ; Peste des petits ruminants virus ; Peste-des-petits-ruminants virus - genetics ; Peste-des-petits-ruminants virus - immunology ; Plasmids ; Ruminantia ; Semliki Forest virus ; Semliki forest virus - genetics ; Suicidal DNA vaccine ; Vaccines, DNA - administration &amp; dosage ; Vaccines, DNA - genetics ; Vaccines, DNA - immunology ; Viral Fusion Proteins - genetics ; Viral Fusion Proteins - immunology</subject><ispartof>Journal of virological methods, 2015-12, Vol.225, p.35-40</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-c401t-d69787d7d45f1390bf34b0b35ef06c6338bc6cdba8a2b9fbb68091b3d2c9a15d3</citedby><cites>FETCH-LOGICAL-c401t-d69787d7d45f1390bf34b0b35ef06c6338bc6cdba8a2b9fbb68091b3d2c9a15d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0166093415003067$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,65309</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26343487$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Yong</creatorcontrib><creatorcontrib>Yue, Xiaolin</creatorcontrib><creatorcontrib>Jin, Hongyan</creatorcontrib><creatorcontrib>Liu, Guangqing</creatorcontrib><creatorcontrib>Pan, Ling</creatorcontrib><creatorcontrib>Wang, Guijun</creatorcontrib><creatorcontrib>Guo, Hao</creatorcontrib><creatorcontrib>Li, Gang</creatorcontrib><creatorcontrib>Li, Yongdong</creatorcontrib><title>A suicidal DNA vaccine expressing the fusion protein of peste des petits ruminants virus induces both humoral and cell-mediated immune responses in mice</title><title>Journal of virological methods</title><addtitle>J Virol Methods</addtitle><description>•A suicidal DNA vaccine against peste des petits ruminants virus (PPRV) was developed.•The suicidal DNA vaccine could express Fusion protein in transfected BHK-21 cells.•Specific humoral and cell-mediated immune responses were induced in immunized mice.•The results provide a new strategy to the development of vaccines against PPRV. Peste des petits ruminants (PPR), a highly contagious disease induced by PPR virus (PPRV), affects sheep and goats. PPRV fusion (F) protein is important for the induction of immune responses against PPRV. We constructed a Semliki Forest virus (SFV) replicon-vectored DNA vaccine (“suicidal DNA vaccine”) and evaluated its immunogenicity in BALB/c mice. The F gene of PPRV was cloned and inserted into the SFV replicon-based vector pSCA1. The antigenicity of the resultant plasmid pSCA1/F was identified by indirect immunofluorescence and western blotting. BALB/c mice were then intramuscularly injected with pSCA1/F three times at 14-d intervals. Specific antibodies and virus-neutralizing antibodies against PPRV were quantified by indirect ELISA and microneutralization tests, respectively. Cell-mediated immune responses were examined by cytokine and lymphocyte proliferation assays. 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This could be considered as a novel strategy for vaccine development against PPR.</description><subject>Animals</subject><subject>Antibodies, Neutralizing - blood</subject><subject>Antibodies, Viral - blood</subject><subject>Cell Proliferation</subject><subject>Cloning, Molecular</subject><subject>Cytokines - secretion</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Fusion protein</subject><subject>Immune response</subject><subject>Immunization Schedule</subject><subject>Injections, Intramuscular</subject><subject>Leukocytes, Mononuclear - immunology</subject><subject>Mice, Inbred BALB C</subject><subject>Neutralization Tests</subject><subject>Peste des petits ruminants virus</subject><subject>Peste-des-petits-ruminants virus - genetics</subject><subject>Peste-des-petits-ruminants virus - immunology</subject><subject>Plasmids</subject><subject>Ruminantia</subject><subject>Semliki Forest virus</subject><subject>Semliki forest virus - genetics</subject><subject>Suicidal DNA vaccine</subject><subject>Vaccines, DNA - administration &amp; dosage</subject><subject>Vaccines, DNA - genetics</subject><subject>Vaccines, DNA - immunology</subject><subject>Viral Fusion Proteins - genetics</subject><subject>Viral Fusion Proteins - immunology</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNUU1v1DAUtBAVXQp_ofKRS4IdJ058Y1VKi1TBpZwtf7ywXm3s4I8V_BN-Ll5tyxVOHskzb96bQeiakpYSyt_v2_3RxbBAbjtCh5aIlhD6Am3oNIqGiKl_iTaVyCtm_SV6ndKeEDKMjL1Clx1nPeuncYN-b3EqzjirDvjjly0-KmOcBww_1wgpOf8d5x3guSQXPF5jyOA8DjNeIWXAFlJF2eWEY1mcV76iulhJ2HlbTP3WIe_wriwhVgvlLTZwODQLWKcyWOyWpVS_arYGn-Ckw4sz8AZdzOqQ4O3Te4W-fbp9vLlvHr7efb7ZPjSmJzQ3lotxGu1o-2GmTBA9s14TzQaYCTecsUkbbqxWk-q0mLXmExFUM9sZoehg2RV6d55bb_tR6lFycem0ovIQSpJ07EYu-CiG_6F2optqAZXKz1QTQ0oRZrlGt6j4S1IiTwXKvXwuUJ4KlETIWmAVXj95FF0z-it7bqwSPpwJUEM5OogyGQfe1DwjmCxtcP_y-APEoLN3</recordid><startdate>20151201</startdate><enddate>20151201</enddate><creator>Wang, Yong</creator><creator>Yue, Xiaolin</creator><creator>Jin, Hongyan</creator><creator>Liu, Guangqing</creator><creator>Pan, Ling</creator><creator>Wang, Guijun</creator><creator>Guo, Hao</creator><creator>Li, Gang</creator><creator>Li, Yongdong</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><scope>7T5</scope><scope>7TM</scope><scope>7U9</scope><scope>H94</scope></search><sort><creationdate>20151201</creationdate><title>A suicidal DNA vaccine expressing the fusion protein of peste des petits ruminants virus induces both humoral and cell-mediated immune responses in mice</title><author>Wang, Yong ; 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dosage</topic><topic>Vaccines, DNA - genetics</topic><topic>Vaccines, DNA - immunology</topic><topic>Viral Fusion Proteins - genetics</topic><topic>Viral Fusion Proteins - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Yong</creatorcontrib><creatorcontrib>Yue, Xiaolin</creatorcontrib><creatorcontrib>Jin, Hongyan</creatorcontrib><creatorcontrib>Liu, Guangqing</creatorcontrib><creatorcontrib>Pan, Ling</creatorcontrib><creatorcontrib>Wang, Guijun</creatorcontrib><creatorcontrib>Guo, Hao</creatorcontrib><creatorcontrib>Li, Gang</creatorcontrib><creatorcontrib>Li, Yongdong</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><collection>Immunology Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Yong</au><au>Yue, Xiaolin</au><au>Jin, Hongyan</au><au>Liu, Guangqing</au><au>Pan, Ling</au><au>Wang, Guijun</au><au>Guo, Hao</au><au>Li, Gang</au><au>Li, Yongdong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A suicidal DNA vaccine expressing the fusion protein of peste des petits ruminants virus induces both humoral and cell-mediated immune responses in mice</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>2015-12-01</date><risdate>2015</risdate><volume>225</volume><spage>35</spage><epage>40</epage><pages>35-40</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><abstract>•A suicidal DNA vaccine against peste des petits ruminants virus (PPRV) was developed.•The suicidal DNA vaccine could express Fusion protein in transfected BHK-21 cells.•Specific humoral and cell-mediated immune responses were induced in immunized mice.•The results provide a new strategy to the development of vaccines against PPRV. Peste des petits ruminants (PPR), a highly contagious disease induced by PPR virus (PPRV), affects sheep and goats. PPRV fusion (F) protein is important for the induction of immune responses against PPRV. We constructed a Semliki Forest virus (SFV) replicon-vectored DNA vaccine (“suicidal DNA vaccine”) and evaluated its immunogenicity in BALB/c mice. The F gene of PPRV was cloned and inserted into the SFV replicon-based vector pSCA1. The antigenicity of the resultant plasmid pSCA1/F was identified by indirect immunofluorescence and western blotting. BALB/c mice were then intramuscularly injected with pSCA1/F three times at 14-d intervals. Specific antibodies and virus-neutralizing antibodies against PPRV were quantified by indirect ELISA and microneutralization tests, respectively. Cell-mediated immune responses were examined by cytokine and lymphocyte proliferation assays. The pSCA1/F expressed F protein in vitro and induced specific and neutralizing antibody production, and lymphocyte proliferation in mice. Mice vaccinated with pSCA1/F had increased IL-2 and IL-10 levels after 24-h post first immunization. IFN-γ and TNF-α levels increased from that time point and gradually decreased thereafter. Thus, the Semliki Forest virus replicon-vectored DNA vaccine expressing the F protein of PPRV induced both humoral and cell-mediated immune responses in mice. This could be considered as a novel strategy for vaccine development against PPR.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>26343487</pmid><doi>10.1016/j.jviromet.2015.09.001</doi><tpages>6</tpages></addata></record>
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subjects Animals
Antibodies, Neutralizing - blood
Antibodies, Viral - blood
Cell Proliferation
Cloning, Molecular
Cytokines - secretion
Enzyme-Linked Immunosorbent Assay
Fusion protein
Immune response
Immunization Schedule
Injections, Intramuscular
Leukocytes, Mononuclear - immunology
Mice, Inbred BALB C
Neutralization Tests
Peste des petits ruminants virus
Peste-des-petits-ruminants virus - genetics
Peste-des-petits-ruminants virus - immunology
Plasmids
Ruminantia
Semliki Forest virus
Semliki forest virus - genetics
Suicidal DNA vaccine
Vaccines, DNA - administration & dosage
Vaccines, DNA - genetics
Vaccines, DNA - immunology
Viral Fusion Proteins - genetics
Viral Fusion Proteins - immunology
title A suicidal DNA vaccine expressing the fusion protein of peste des petits ruminants virus induces both humoral and cell-mediated immune responses in mice
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