Study of the Immunogenicity of the VP2 Protein of Canine Parvovirus Produced Using an Improved Baculovirus Expression System

Overview

Journal BMC Vet Res

Publisher Biomed Central

Specialty Veterinary Medicine

Date 2020 Jun 20

PMID 32552679

Citations 6

Authors

Dao Chang

Yangkun Liu

Yangyang Chen

Xiaomin Hu

Andrey Burov

Alexey Puzyr

Vladimir Bondar

Lunguang Yao

Affiliations

Soon will be listed here.

Abstract

Background: Canine parvovirus (CPV) is now recognized as a serious threat to the dog breeding industry worldwide. Currently used CPV vaccines all have their specific drawbacks, prompting a search for alternative safe and effective vaccination strategies such as subunit vaccine. VP2 protein is the major antigen targeted for developing CPV subunit vaccine, however, its production in baculovirus expression system remains challenging due to the insufficient yield. Therefore, our study aims to increase the VP2 protein production by using an improved baculovirus expression system and to evaluate the immunogenicity of the purified VP2 protein in mice.

Results: The results showed that high-level expression of the full length VP2 protein was achieved using our modified baculovirus expression system. The recombinant virus carrying two copies of VP2 gene showed the highest expression level, with a productivity of 186 mg/L, which is about 1.4-1.6 fold that of the recombinant viruses carrying only one copy. The purified protein reacted with Mouse anti-His tag monoclonal antibody and Rabbit anti-VP2 polyclonal antibody. BALB/c mice were intramuscularly immunized with purified VP2 protein twice at 2 week intervals. After vaccination, VP2 protein could induce the mice produce high level of hemagglutination inhibition antibodies.

Conclusions: Full length CPV VP2 protein was expressed at high level and purified efficiently. Moreover, it stimulated mice to produce high level of antibodies with hemmaglutination inhibition properties. The VP2 protein expressed in this study could be used as a putative economic and efficient subunit vaccine against CPV infection.

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References

Berger I, Fitzgerald D, Richmond T . Baculovirus expression system for heterologous multiprotein complexes. Nat Biotechnol. 2004; 22(12):1583-7. DOI: 10.1038/nbt1036. View

Elia G, Desario C, Pezzoni G, Camero M, Brocchi E, Decaro N . Recombinant ELISA using baculovirus-expressed VP2 for detection of antibodies against canine parvovirus. J Virol Methods. 2012; 184(1-2):98-102. DOI: 10.1016/j.jviromet.2012.05.025. View

Feng H, Hu G, Wang H, Liang M, Liang H, Guo H . Canine parvovirus VP2 protein expressed in silkworm pupae self-assembles into virus-like particles with high immunogenicity. PLoS One. 2014; 9(1):e79575. PMC: 3894932. DOI: 10.1371/journal.pone.0079575. View

Wu Z, Sun H, Zhang X . [Immunoadjuvant effect of the extracellular domain of canine CTLA-4 on the VP2 protein of canine parvovirus]. Wei Sheng Wu Xue Bao. 2008; 48(3):369-74. View

Park J, Choi B, Vijayachandran L, Ayyappan V, Chong C, Lee K . Immunodetection of Canine Parvovirus (CPV) in clinical samples by polyclonal antisera against CPV-VP2 protein expressed in Esherichia coli as an antigen. J Virol Methods. 2007; 146(1-2):281-7. DOI: 10.1016/j.jviromet.2007.07.021. View

Mira F, Purpari G, Lorusso E, Di Bella S, Gucciardi F, Desario C . Introduction of Asian canine parvovirus in Europe through dog importation. Transbound Emerg Dis. 2017; 65(1):16-21. PMC: 7169687. DOI: 10.1111/tbed.12747. View

Singh P, Destito G, Schneemann A, Manchester M . Canine parvovirus-like particles, a novel nanomaterial for tumor targeting. J Nanobiotechnology. 2006; 4:2. PMC: 1386698. DOI: 10.1186/1477-3155-4-2. View

Rehm B . Bioengineering towards self-assembly of particulate vaccines. Curr Opin Biotechnol. 2017; 48:42-53. DOI: 10.1016/j.copbio.2017.03.018. View

Xu J, Guo H, Wei Y, Dong H, Han S, Ao D . Self-assembly of virus-like particles of canine parvovirus capsid protein expressed from Escherichia coli and application as virus-like particle vaccine. Appl Microbiol Biotechnol. 2014; 98(8):3529-38. DOI: 10.1007/s00253-013-5485-6. View

10.

Decaro N, Desario C, Parisi A, Martella V, Lorusso A, Miccolupo A . Genetic analysis of canine parvovirus type 2c. Virology. 2009; 385(1):5-10. DOI: 10.1016/j.virol.2008.12.016. View

11.

Kyratsous C, Silverstein S, DeLong C, Panagiotidis C . Chaperone-fusion expression plasmid vectors for improved solubility of recombinant proteins in Escherichia coli. Gene. 2009; 440(1-2):9-15. PMC: 2683908. DOI: 10.1016/j.gene.2009.03.011. View

12.

Nan L, Liu Y, Ji P, Feng H, Chen C, Wang J . Trigger factor assisted self-assembly of canine parvovirus VP2 protein into virus-like particles in Escherichia coli with high immunogenicity. Virol J. 2018; 15(1):103. PMC: 6008937. DOI: 10.1186/s12985-018-1013-8. View

13.

Kaur J, Kumar A, Kaur J . Strategies for optimization of heterologous protein expression in E. coli: Roadblocks and reinforcements. Int J Biol Macromol. 2017; 106:803-822. DOI: 10.1016/j.ijbiomac.2017.08.080. View

14.

Zhou P, Zeng W, Zhang X, Li S . The genetic evolution of canine parvovirus - A new perspective. PLoS One. 2017; 12(3):e0175035. PMC: 5376324. DOI: 10.1371/journal.pone.0175035. View

15.

Allison A, Organtini L, Zhang S, Hafenstein S, Holmes E, Parrish C . Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. J Virol. 2015; 90(2):753-67. PMC: 4702700. DOI: 10.1128/JVI.02636-15. View

16.

Day M, Horzinek M, Schultz R . WSAVA guidelines for the vaccination of dogs and cats. J Small Anim Pract. 2010; 51(6):1-32. PMC: 7166980. DOI: 10.1111/j.1748-5827.2010.00959a.x. View

17.

Decaro N, Desario C, Campolo M, Elia G, Martella V, Ricci D . Clinical and virological findings in pups naturally infected by canine parvovirus type 2 Glu-426 mutant. J Vet Diagn Invest. 2005; 17(2):133-8. DOI: 10.1177/104063870501700206. View

18.

Gileadi O . Recombinant Protein Expression in E. coli : A Historical Perspective. Methods Mol Biol. 2017; 1586:3-10. DOI: 10.1007/978-1-4939-6887-9_1. View

19.

Feng H, Liang M, Wang H, Zhang T, Zhao P, Shen X . Recombinant canine parvovirus-like particles express foreign epitopes in silkworm pupae. Vet Microbiol. 2011; 154(1-2):49-57. DOI: 10.1016/j.vetmic.2011.06.022. View

20.

Chambers A, Aksular M, Graves L, Irons S, Possee R, King L . Overview of the Baculovirus Expression System. Curr Protoc Protein Sci. 2018; 91:5.4.1-5.4.6. DOI: 10.1002/cpps.47. View