| Virology Journal | |
| Induction of robust immunity response in mice by dual-expression-system-based recombinant baculovirus expressing the capsid protein of porcine circovirus type 2 | |
| Huiying Fan2  Ming Liao2  Wenyao Lin2  Tiezhu Tong1  Jie Zhang2  Xiaoliang Cheng2  Yu Ye2  | |
| [1] Huizhou Entry-Exit Inspection and Quarantine Bureau, Huizhou 516001, China;College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China | |
| 关键词: Immune response; Dual-expression-system; Cap protein; Vaccine; Porcine circovirus type 2; | |
| Others : 820448 DOI : 10.1186/1743-422X-10-316 |
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| received in 2013-06-13, accepted in 2013-10-22, 发布年份 2013 | |
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【 摘 要 】
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.
【 授权许可】
2013 Ye et al.; licensee BioMed Central Ltd.
【 预 览 】
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| 20140712042853600.pdf | 768KB | ||
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| Figure 5. | 95KB | Image | |
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| Figure 2. | 43KB | Image | |
| Figure 1. | 44KB | Image |
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【 参考文献 】
- [1]Opriessnig T, Patterson AR, Elsener J, Meng XJ, Halbur PG: Influence of maternal antibodies on efficacy of porcine circovirus type 2 (PCV2) vaccination to protect pigs from experimental infection with PCV2. Clin Vaccine Immunol 2008, 15:397-401.
- [2]Fort M, Sibila M, Perez-Martin E, Nofrarias M, Mateu E, Segales J: One dose of a porcine circovirus 2 (PCV2) sub-unit vaccine administered to 3-week-old conventional piglets elicits cell-mediated immunity and significantly reduces PCV2 viremia in an experimental model. Vaccine 2009, 27:4031-4037.
- [3]Lyoo K, Joo H, Caldwell B, Kim H, Davies PR, Torrison J: Comparative efficacy of three commercial PCV2 vaccines in conventionally reared pigs. Vet J 2011, 189:58-62.
- [4]Koch G, van Roozelaar DJ, Verschueren CA, van der Eb AJ, Noteborn MH: Immunogenic and protective properties of chicken anaemia virus proteins expressed by baculovirus. Vaccine 1995, 13:763-770.
- [5]Krakowka S, Ellis JA, McNeilly F, Ringler S, Rings DM, Allan G: Activation of the immune system is the pivotal event in the production of wasting disease in pigs infected with porcine circovirus-2 (PCV-2). Vet Pathol 2001, 38:31-42.
- [6]Fan H, Ju C, Tong T, Huang H, Lv J, Chen H: Immunogenicity of empty capsids of porcine circovius type 2 produced in insect cells. Vet Res Commun 2007, 31:487-496.
- [7]Hofmann C, Sandig V, Jennings G, Rudolph M, Schlag P, Strauss M: Efficient gene transfer into human hepatocytes by baculovirus vectors. Proc Natl Acad Sci U S A 1995, 92:10099-10103.
- [8]Abe T, Takahashi H, Hamazaki H, Miyano-Kurosaki N, Matsuura Y, Takaku H: Baculovirus induces an innate immune response and confers protection from lethal influenza virus infection in mice. J Immunol 2003, 171:1133-1139.
- [9]Fan H, Pan Y, Fang L, Wang D, Wang S, Jiang Y, Chen H, Xiao S: Construction and immunogenicity of recombinant pseudotype baculovirus expressing the capsid protein of porcine circovirus type 2 in mice. J Virol Methods 2008, 150:21-26.
- [10]Pan Y, Fang L, Fan H, Luo R, Zhao Q, Chen H, Xiao S: Antitumor effects of a recombinant pseudotype baculovirus expressing Apoptin in vitro and in vivo. Int J Cancer 2010, 126:2741-2751.
- [11]Barsoum J, Brown R, McKee M, Boyce FM: Efficient transduction of mammalian cells by a recombinant baculovirus having the vesicular stomatitis virus G glycoprotein. Hum Gene Ther 1997, 8:2011-2018.
- [12]Sarkis C, Serguera C, Petres S, Buchet D, Ridet JL, Edelman L, Mallet J: Efficient transduction of neural cells in vitro and in vivo by a baculovirus-derived vector. Proc Natl Acad Sci U S A 2000, 97:14638-14643.
- [13]Lu L, Yu L, Kwang J: Baculovirus surface-displayed hemagglutinin of H5N1 influenza virus sustains its authentic cleavage, hemagglutination activity, and antigenicity. Biochem Biophys Res Commun 2007, 358:404-409.
- [14]Feng Q, Liu Y, Qu X, Deng H, Ding M, Lau TL, Yu AC, Chen J: Baculovirus surface display of SARS coronavirus (SARS-CoV) spike protein and immunogenicity of the displayed protein in mice models. DNA Cell Biol 2006, 25:668-673.
- [15]Xu XG, Liu HJ: Baculovirus surface display of E2 envelope glycoprotein of classical swine fever virus and immunogenicity of the displayed proteins in a mouse model. Vaccine 2008, 26:5455-5460.
- [16]Tang XC, Lu HR, Ross TM: Hemagglutinin displayed baculovirus protects against highly pathogenic influenza. Vaccine 2010, 28:6821-6831.
- [17]Ellis J, Hassard L, Clark E, Harding J, Allan G, Willson P, Strokappe J, Martin K, McNeilly F, Meehan B, et al.: Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome. Can Vet J 1998, 39:44-51.
- [18]Morozov I, Sirinarumitr T, Sorden SD, Halbur PG, Morgan MK, Yoon KJ, Paul PS: Detection of a novel strain of porcine circovirus in pigs with postweaning multisystemic wasting syndrome. J Clin Microbiol 1998, 36:2535-2541.
- [19]Gillespie J, Opriessnig T, Meng XJ, Pelzer K, Buechner-Maxwell V: Porcine circovirus type 2 and porcine circovirus-associated disease. J Vet Intern Med 2009, 23:1151-1163.
- [20]Kamstrup S, Barfoed AM, Frimann TH, Ladekjaer-Mikkelsen AS, Botner A: Immunisation against PCV2 structural protein by DNA vaccination of mice. Vaccine 2004, 22:1358-1361.
- [21]Kost TA, Condreay JP, Jarvis DL: Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nat Biotechnol 2005, 23:567-575.
- [22]Chen CY, Liu HJ, Tsai CP, Chung CY, Shih YS, Chang PC, Chiu YT, Hu YC: Baculovirus as an avian influenza vaccine vector: differential immune responses elicited by different vector forms. Vaccine 2010, 28:7644-7651.
- [23]Abe T, Hemmi H, Miyamoto H, Moriishi K, Tamura S, Takaku H, Akira S, Matsuura Y: Involvement of the Toll-like receptor 9 signaling pathway in the induction of innate immunity by baculovirus. J Virol 2005, 79:2847-2858.
- [24]Abe T, Kaname Y, Wen X, Tani H, Moriishi K, Uematsu S, Takeuchi O, Ishii KJ, Kawai T, Akira S, Matsuura Y: Baculovirus induces type I interferon production through toll-like receptor-dependent and -independent pathways in a cell-type-specific manner. J Virol 2009, 83:7629-7640.
- [25]Strauss R, Huser A, Ni S, Tuve S, Kiviat N, Sow PS, Hofmann C, Lieber A: Baculovirus-based vaccination vectors allow for efficient induction of immune responses against plasmodium falciparum circumsporozoite protein. Mol Ther 2007, 15:193-202.
- [26]Meerts P, Misinzo G, Nauwynck HJ: Enhancement of porcine circovirus 2 replication in porcine cell lines by IFN-gamma before and after treatment and by IFN-alpha after treatment. J Interferon Cytokine Res 2005, 25:684-693.
- [27]Shen HG, Beach NM, Huang YW, Halbur PG, Meng XJ, Opriessnig T: Comparison of commercial and experimental porcine circovirus type 2 (PCV2) vaccines using a triple challenge with PCV2, porcine reproductive and respiratory syndrome virus (PRRSV), and porcine parvovirus (PPV). Vaccine 2010, 28:5960-5966.
- [28]Hu YC: Baculovirus as a highly efficient expression vector in insect and mammalian cells. Acta Pharmacol Sin 2005, 26:405-416.
- [29]Abe T, Matsuura Y: Host innate immune responses induced by baculovirus in mammals. Curr Gene Ther 2010, 10:226-231.
- [30]Zhu Y, Lau A, Lau J, Jia Q, Karuppannan AK, Kwang J: Enhanced replication of porcine circovirus type 2 (PCV2) in a homogeneous subpopulation of PK15 cell line. Virology 2007, 369:423-430.
- [31]van Oers MM: Opportunities and challenges for the baculovirus expression system. J Invertebr Pathol 2011, 107(Suppl):S3-S15.
- [32]Genmei L, Manlin L, Ruiai C, Hongliang H, Dangshuai P: Construction and immunogenicity of recombinant adenovirus expressing ORF2 of PCV2 and porcine IFN gamma. Vaccine 2011, 29:8677-8682.
- [33]Wang YP, Liu D, Guo LJ, Tang QH, Wei YW, Wu HL, Liu JB, Li SB, Huang LP, Liu CM: Enhanced protective immune response to PCV2 subunit vaccine by co-administration of recombinant porcine IFN-gamma in mice. Vaccine 2013, 31:833-838.
- [34]Roques E, Girard A, Gagnon CA, Archambault D: Antibody responses induced in mice immunized with recombinant adenovectors expressing chimeric proteins of various porcine pathogens. Vaccine 2013, 31:2698-2704.
- [35]O'Reilly DR, Miller LK, Luckow VA: Baculovirus expression vectors: a laboratory manual. New York: Oxford University Press; 1994.
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