Cargando…

A DNA Vaccine Encoding the VAA Gene of Vibrio anguillarum Induces a Protective Immune Response in Flounder

Vibrio anguillarum is a pathogenic bacterium that infects flounder resulting in significant losses in the aquaculture industry. The VAA protein previously identified in flounder is associated with a role in immune protection within these fish. In the present study, a recombinant DNA plasmid encoding...

Descripción completa

Detalles Bibliográficos
Autores principales: Xing, Jing, Xu, Hongsen, Tang, Xiaoqian, Sheng, Xiuzhen, Zhan, Wenbin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6435001/
https://www.ncbi.nlm.nih.gov/pubmed/30941134
http://dx.doi.org/10.3389/fimmu.2019.00499
_version_ 1783406581038186496
author Xing, Jing
Xu, Hongsen
Tang, Xiaoqian
Sheng, Xiuzhen
Zhan, Wenbin
author_facet Xing, Jing
Xu, Hongsen
Tang, Xiaoqian
Sheng, Xiuzhen
Zhan, Wenbin
author_sort Xing, Jing
collection PubMed
description Vibrio anguillarum is a pathogenic bacterium that infects flounder resulting in significant losses in the aquaculture industry. The VAA protein previously identified in flounder is associated with a role in immune protection within these fish. In the present study, a recombinant DNA plasmid encoding the VAA gene of V. anguillarum was constructed and its potential as a DNA vaccine, to prevent the infection of V. anguillarum in flounder fish, investigated. We verified the expression of the VAA protein both in vitro in cell lines and in vivo in flounder fish. The protective effects of pcDNA3.1-VAA (pVAA) were analyzed by determination of the percentage of sIgM(+), CD4-1(+), CD4-2(+), CD8β(+) lymphocytes, and the production of VAA-specific antibodies in flounder following their immunization with the DNA vaccine. Histopathological changes in immune related tissues, bacterial load, and relative percentage survival rates of flounder post-challenge with V. anguillarum, were all investigated to assess the efficacy of the pVAA DNA vaccine candidate. Fish intramuscularly immunized with pVAA showed a significant increase in CD4-1(+), CD4-2(+), and CD8β(+) T lymphocytes at days 9, 11, and 14 post-vaccination, reaching peak T-cell levels at days 11 or 14 post-immunization. The percentage of sIgM(+) lymphocytes reached peak levels at weeks 4–5 post-immunization. Specific anti-V. anguillarum or anti-rVAA antibodies were induced in inoculated fish at days 28–35 post-immunization. The liver of vaccinated flounder exhibited only slight histopathological changes compared with a significant pathology observed in control immunized fish. Additionally, a lower bacterial burden in the liver, spleen, and kidney were observed in pVAA protected fish in response to bacterial challenge, compared with pcDNA3.1 vector control injected fish. Moreover, the pVAA vaccine confers a relative percentage survival of 50.00% following V. anguillarum infection. In summary, this is the first study indicating an initial induction of the T lymphocyte response, followed by B lymphocyte induction of specific antibodies as a result of DNA immunization of flounder. This signifies the important potential of pVAA as a DNA vaccine candidate for the control of V. anguillarum infection.
format Online
Article
Text
id pubmed-6435001
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-64350012019-04-02 A DNA Vaccine Encoding the VAA Gene of Vibrio anguillarum Induces a Protective Immune Response in Flounder Xing, Jing Xu, Hongsen Tang, Xiaoqian Sheng, Xiuzhen Zhan, Wenbin Front Immunol Immunology Vibrio anguillarum is a pathogenic bacterium that infects flounder resulting in significant losses in the aquaculture industry. The VAA protein previously identified in flounder is associated with a role in immune protection within these fish. In the present study, a recombinant DNA plasmid encoding the VAA gene of V. anguillarum was constructed and its potential as a DNA vaccine, to prevent the infection of V. anguillarum in flounder fish, investigated. We verified the expression of the VAA protein both in vitro in cell lines and in vivo in flounder fish. The protective effects of pcDNA3.1-VAA (pVAA) were analyzed by determination of the percentage of sIgM(+), CD4-1(+), CD4-2(+), CD8β(+) lymphocytes, and the production of VAA-specific antibodies in flounder following their immunization with the DNA vaccine. Histopathological changes in immune related tissues, bacterial load, and relative percentage survival rates of flounder post-challenge with V. anguillarum, were all investigated to assess the efficacy of the pVAA DNA vaccine candidate. Fish intramuscularly immunized with pVAA showed a significant increase in CD4-1(+), CD4-2(+), and CD8β(+) T lymphocytes at days 9, 11, and 14 post-vaccination, reaching peak T-cell levels at days 11 or 14 post-immunization. The percentage of sIgM(+) lymphocytes reached peak levels at weeks 4–5 post-immunization. Specific anti-V. anguillarum or anti-rVAA antibodies were induced in inoculated fish at days 28–35 post-immunization. The liver of vaccinated flounder exhibited only slight histopathological changes compared with a significant pathology observed in control immunized fish. Additionally, a lower bacterial burden in the liver, spleen, and kidney were observed in pVAA protected fish in response to bacterial challenge, compared with pcDNA3.1 vector control injected fish. Moreover, the pVAA vaccine confers a relative percentage survival of 50.00% following V. anguillarum infection. In summary, this is the first study indicating an initial induction of the T lymphocyte response, followed by B lymphocyte induction of specific antibodies as a result of DNA immunization of flounder. This signifies the important potential of pVAA as a DNA vaccine candidate for the control of V. anguillarum infection. Frontiers Media S.A. 2019-03-19 /pmc/articles/PMC6435001/ /pubmed/30941134 http://dx.doi.org/10.3389/fimmu.2019.00499 Text en Copyright © 2019 Xing, Xu, Tang, Sheng and Zhan. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Xing, Jing
Xu, Hongsen
Tang, Xiaoqian
Sheng, Xiuzhen
Zhan, Wenbin
A DNA Vaccine Encoding the VAA Gene of Vibrio anguillarum Induces a Protective Immune Response in Flounder
title A DNA Vaccine Encoding the VAA Gene of Vibrio anguillarum Induces a Protective Immune Response in Flounder
title_full A DNA Vaccine Encoding the VAA Gene of Vibrio anguillarum Induces a Protective Immune Response in Flounder
title_fullStr A DNA Vaccine Encoding the VAA Gene of Vibrio anguillarum Induces a Protective Immune Response in Flounder
title_full_unstemmed A DNA Vaccine Encoding the VAA Gene of Vibrio anguillarum Induces a Protective Immune Response in Flounder
title_short A DNA Vaccine Encoding the VAA Gene of Vibrio anguillarum Induces a Protective Immune Response in Flounder
title_sort dna vaccine encoding the vaa gene of vibrio anguillarum induces a protective immune response in flounder
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6435001/
https://www.ncbi.nlm.nih.gov/pubmed/30941134
http://dx.doi.org/10.3389/fimmu.2019.00499
work_keys_str_mv AT xingjing adnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder
AT xuhongsen adnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder
AT tangxiaoqian adnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder
AT shengxiuzhen adnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder
AT zhanwenbin adnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder
AT xingjing dnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder
AT xuhongsen dnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder
AT tangxiaoqian dnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder
AT shengxiuzhen dnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder
AT zhanwenbin dnavaccineencodingthevaageneofvibrioanguillaruminducesaprotectiveimmuneresponseinflounder