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Induction of robust immunity response in mice by dual-expression-system-based recombinant baculovirus expressing the capsid protein of porcine circovirus type 2.

Ye Y, Cheng X, Zhang J, Tong T, Lin W, Liao M, Fan H - Virol. J. (2013)

Bottom Line: Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS), an emerging swine disease that causes progressive weight loss, dyspnea, tachypnea, anemia, jaundice, and diarrhea in piglets.In this study, we successfully constructed a dual-expression-system-based recombinant baculovirus BV-GD-ORF2, which can display the PCV2 capsid (Cap) protein and VSV-G protein on the viral envelope and also expressing Cap protein on transduced mammalian cells, thereby functioning as both a subunit and a DNA vaccine.The vaccination of mice with recombinant baculovirus BV-GD-ORF2 successfully induced robust Cap-protein-specific humoral and cellular immune responses.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Animal Vaccine Development, Ministry of Agriculture, Guangzhou 510642, China. fanhy@scau.edu.cn.

ABSTRACT

Background: Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS), an emerging swine disease that causes progressive weight loss, dyspnea, tachypnea, anemia, jaundice, and diarrhea in piglets. Although baculovirus is an enveloped virus that infects insects in nature, it has emerged as a vaccine vector, and we used it to develop a novel candidate vaccine for a preventive or therapeutic strategy to control PCV2 infections.

Methods: Immunoblotting analysis of recombinant baculovirus and immunofluorescent staining of baculovirus-infected cells were followed using anti-ORF2 monoclonal antibodies. The BALB/c mice were immunized intramuscularly with this baculovirus. The titers of antibodies were mensurated with a Cap-protein-specific enzyme-linked immunosorbent assay (ELISA) and a serum neutralization assay. The IFN-γ response in splenocytes harvested from immunized mice was measured by ELISA. Student's t-test was used to compare immune responses of different groups.

Results: In this study, we successfully constructed a dual-expression-system-based recombinant baculovirus BV-GD-ORF2, which can display the PCV2 capsid (Cap) protein and VSV-G protein on the viral envelope and also expressing Cap protein on transduced mammalian cells, thereby functioning as both a subunit and a DNA vaccine. After infection, the Cap protein was expressed and displayed on the viral surface, as demonstrated with an indirect fluorescence assay and immunoblotting. The vaccination of mice with recombinant baculovirus BV-GD-ORF2 successfully induced robust Cap-protein-specific humoral and cellular immune responses.

Conclusions: Our findings collectively demonstrate that the recombinant baculovirus BV-GD-ORF2 is a potential vaccine against PCV2 infections.

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Characterization of BV-GD-ORF2-infected Sf9 cells. Syncytium formation in Sf9 cells infected with BV-GD-ORF2 as indicated by the white arrow (a), but not in the mock-infected Sf9 cells (b). The images were captured at 72 h posttransfection. Original magnification, ×100.
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Figure 2: Characterization of BV-GD-ORF2-infected Sf9 cells. Syncytium formation in Sf9 cells infected with BV-GD-ORF2 as indicated by the white arrow (a), but not in the mock-infected Sf9 cells (b). The images were captured at 72 h posttransfection. Original magnification, ×100.

Mentions: The recombinant baculovirus BV-GD-ORF2 was constructed as described in the Methods (Figure 1). The infection of Sf9 cells with BV-GD-ORF2 caused in extensive cell-cell fusion (Figure 2). This phenotype is attributable to the very high expression level of VSV-G protein, which has membrane-fusion activity, under the control of the polyhedrin promoter (PPH).


Induction of robust immunity response in mice by dual-expression-system-based recombinant baculovirus expressing the capsid protein of porcine circovirus type 2.

Ye Y, Cheng X, Zhang J, Tong T, Lin W, Liao M, Fan H - Virol. J. (2013)

Characterization of BV-GD-ORF2-infected Sf9 cells. Syncytium formation in Sf9 cells infected with BV-GD-ORF2 as indicated by the white arrow (a), but not in the mock-infected Sf9 cells (b). The images were captured at 72 h posttransfection. Original magnification, ×100.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231451&req=5

Figure 2: Characterization of BV-GD-ORF2-infected Sf9 cells. Syncytium formation in Sf9 cells infected with BV-GD-ORF2 as indicated by the white arrow (a), but not in the mock-infected Sf9 cells (b). The images were captured at 72 h posttransfection. Original magnification, ×100.
Mentions: The recombinant baculovirus BV-GD-ORF2 was constructed as described in the Methods (Figure 1). The infection of Sf9 cells with BV-GD-ORF2 caused in extensive cell-cell fusion (Figure 2). This phenotype is attributable to the very high expression level of VSV-G protein, which has membrane-fusion activity, under the control of the polyhedrin promoter (PPH).

Bottom Line: Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS), an emerging swine disease that causes progressive weight loss, dyspnea, tachypnea, anemia, jaundice, and diarrhea in piglets.In this study, we successfully constructed a dual-expression-system-based recombinant baculovirus BV-GD-ORF2, which can display the PCV2 capsid (Cap) protein and VSV-G protein on the viral envelope and also expressing Cap protein on transduced mammalian cells, thereby functioning as both a subunit and a DNA vaccine.The vaccination of mice with recombinant baculovirus BV-GD-ORF2 successfully induced robust Cap-protein-specific humoral and cellular immune responses.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Animal Vaccine Development, Ministry of Agriculture, Guangzhou 510642, China. fanhy@scau.edu.cn.

ABSTRACT

Background: Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS), an emerging swine disease that causes progressive weight loss, dyspnea, tachypnea, anemia, jaundice, and diarrhea in piglets. Although baculovirus is an enveloped virus that infects insects in nature, it has emerged as a vaccine vector, and we used it to develop a novel candidate vaccine for a preventive or therapeutic strategy to control PCV2 infections.

Methods: Immunoblotting analysis of recombinant baculovirus and immunofluorescent staining of baculovirus-infected cells were followed using anti-ORF2 monoclonal antibodies. The BALB/c mice were immunized intramuscularly with this baculovirus. The titers of antibodies were mensurated with a Cap-protein-specific enzyme-linked immunosorbent assay (ELISA) and a serum neutralization assay. The IFN-γ response in splenocytes harvested from immunized mice was measured by ELISA. Student's t-test was used to compare immune responses of different groups.

Results: In this study, we successfully constructed a dual-expression-system-based recombinant baculovirus BV-GD-ORF2, which can display the PCV2 capsid (Cap) protein and VSV-G protein on the viral envelope and also expressing Cap protein on transduced mammalian cells, thereby functioning as both a subunit and a DNA vaccine. After infection, the Cap protein was expressed and displayed on the viral surface, as demonstrated with an indirect fluorescence assay and immunoblotting. The vaccination of mice with recombinant baculovirus BV-GD-ORF2 successfully induced robust Cap-protein-specific humoral and cellular immune responses.

Conclusions: Our findings collectively demonstrate that the recombinant baculovirus BV-GD-ORF2 is a potential vaccine against PCV2 infections.

Show MeSH
Related in: MedlinePlus