Localization of the VP2 Protein of Canine Parvovirus Type 2 on the Baculovirus Envelop and Its Immunogenicity in a Mouse Model

Chih H. Tsai¹, Jing Y. Wang², Xin G. Xu², De W. Tong², Hsin Y. Lu³, Yi H. Chen³, Ming T. Chiou⁴, Ching D. Chang⁴, Hung J. Liu^5*

¹Graduate Institute of Vaccine Biotechnology, Pingtung.
²College of Veterinary Medicine, Northwest A&F University, Yangling, China.
³Institute of Molecular Biology, National Chung Hsing University, Taichung.
⁴Department of Veterinary Medicine, National Pingtung University of Science and Technology, Neipu.
⁵Agricultural Biotechnology Center, National Chung Hsing University, Taichung.

DOI: 10.4236/ojvm.2012.24028                

Abstract

In this study, the full-length VP2 gene of canine parvovirus type 2 (CPV-2) was cloned into the pBacSC vector which possesses baculovirus transmembrane domain (gp64 TM) gene, baculovirus cytoplasmic domain (gp64 CTD) gene, and green florescence protein (GFP) gene. Baculovirus gp64 TM and gp64 CTD in the pBacSC vector were designed to display heterologous proteins on the baculovirus envelope. After cloning the VP2 gene of CPV-2 into pBacSC vector, the recombinant plasmid pBacSC-VP2 was transformed into E. coli DH10Bac competent cells to form recombinant bacmid DNA. One recombinant baculovirus BacSC-VP2 that expresses the VP2 protein of CPV-2 was obtained. Confocal microscopy and immunogold electron microscopy were used to verify whether VP2 expressing on baculovirus envelope or cell membrane. Immunization of BALB/c mice with recombinant baculovirus BacSC-VP2, demonstrated that serum from the BacSC-VP2 treated models had higher levels of virus neutralization titers than the control groups. The results show that the recombinant baculovirus BacSC-VP2 can induce a strong immune response in a mouse model, suggesting that the pseudotyped baculovirus BacSC-VP2 can serve as a potential vaccine against CPV infections.

Keywords

Canine Parvovirus Type 2; VP2 Protein; Baculovirus gp64 TM and CTD; Subunit Vaccine

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	U. Truyen, “Evolution of Canine Parvovirus—A Need for New Vaccines?” Veterinary Microbiology, Vol. 117, No. 1, 2006, pp. 9-13. doi:10.1016/j.vetmic.2006.04.003
[2]	M. J. Appel, W. Scott and L. E. Carmichael, “Isolation and Immunization Studies of A Canine Parvo-Like Virus From Dogs With Haemorrhagic Enteritis,” Veterinary Record, Vol. 105, No. 8, 1979, pp. 156-159. doi:10.1136/vr.105.8.156
[3]	W. R. Kelly, “An Enteric Disease of Dogs Reselmbing Feline Panleucopaenia,” Austrian Veterinary Journal, Vol. 54, No. 12, 1978, p. 593. doi:10.1111/j.1751-0813.1978.tb02426.x
[4]	G. Burtonboy, F. Coignoul and N. Delferriere, “Canine Hemorrhagic Enteritis: Detection of Viral Particles by Electron Microscopy,” Archives of Virology, Vol. 61, No. 1-2, 1979, pp. 1-11. doi:10.1007/BF01320586
[5]	N. Decaro, C. Desario, M. Campolo, G. Elia, V. Martella, D. Ricci, E. Lorusso and C. Buonavoglia, “Clinical and Virological Findings In Pups Naturally Infected by Canine Parvovirus Type 2 Glu-426 Mutant,” Journal of Veterinary Diagnostic Investigation, Vol. 17, No. 2, 2005, pp. 133-138. doi:10.1177/104063870501700206
[6]	M. Mochizuki, M. Horiuchi. H. Hiragi. M. C. San Gabriel, N. Yasuda and T. Uno, “Isolation of Canine Parvovirus From a Cat Manifesting Clinical Signs of Feline Panleukopenia,” Journal of Clinical Microbiology, Vol. 34, No. 9, 1996, pp. 2101-2105.
[7]	W. Yuan and C. R. Parrish, “Comparison of Two Single-Chain Antibodies That Neutralize Canine Parvovirus: Analysis of an Antibody-Combining Site and Mechanisms of Neutralization,” Virology, Vol. 269, No. 2, 2000, pp. 471-480. doi:10.1006/viro.2000.0230
[8]	M. Nakamura, Y. Tohya, T. Miyazawa, M. Mochizuki, H. T. Phung, N. Nguyen, L. M. Huynh, L. T. Nguyen, P. N. Nguyen, P. V. Nguyen, N. P. Nguyen and H. Akashi, “A Novel Antigenic Variant of Canine Parvovirus from a Vietnamese Dog,” Archives of Virology, Vol. 149, No. 11, 2004, pp. 2261-2269. doi:10.1007/s00705-004-0367-y
[9]	H. S. Moon, S. A. Lee, S. G. Lee, R. Choi, S. Y. Jeoung, D. Kim and C. Hyun, “Comparison of the Pathogenicity in Three Different Korean Canine Parvovirus 2 (CPV-2) Isolates,” Veterinary Microbiology, Vol. 131, No. 1-2, 2008, pp. 47-56. doi:10.1016/j.vetmic.2008.02.016
[10]	C. Buonavoglia, V. Martella. A. Pratelli. M. Tempesta. A. Cavalli, D. Buonavoglia. G. Bozzo, G. Elia, N. Decaro and L. Carmichael, “Evidence for Evolution of Canine Parvovirus Type 2 in Italy,” Journal of General Virology, 82, Pt 12, 2001, pp. 3021-3025.
[11]	Y. H. Lin, L. H., Lee, W. L. Shih, Y. C. Hu and H. J. Liu, “Baculovirus Surface Display of σC and σB Proteins of Avian Reovirus and Immunogenicity of the Displayed Proteins in a Mouse Model,” Vaccine, Vol. 26, No. 50, 2008, pp. 6361-6367. doi:10.1016/j.vaccine.2008.09.008
[12]	D. G. Yang, Y. C. Chung, Y. K. Lai, H. J. Liu and Y. C. Hu, “Avian Influenza Virus Hemagglutinin Display on Baculovirus Envelope: Cytoplasmic Domain Affects Virus Properties and Vaccine Potential,” Molecular Therapy, Vol.15, No. 5, 2007. pp. 989-996. doi:10.1038/mt.sj.6300131
[13]	X. G. Xu, M. T., Chiou, Y. M. Zhang, D. W. Tong, J. H. Hu, M. T. Zhang and H. J. Liu, “Baculovirus Surface Display of E (rns) Envelope Glycoprotein of Classical Swine Fever Virus,” Journal of Virological Methods, Vol. 153, No. 2, 2008, pp. 149-155. doi:10.1016/j.jviromet.2008.07.019
[14]	C. R. Parrish, C. F. Aquadro, M. L. Strassheim, J. F. Evermann and J. Y. Sgro, “Mohammed HO. Rapid Antigenic-Type Replacement and DNA Sequence Evolution of Canine Parvovirus,” Journal of Virology, Vol. 65, No. 12, 1991, pp. 6544-6552.
[15]	N. Decaro, G. Elia, C. Desario, S. Roperto, V. Martella, M. Campolo, A. Lorusso, A. Cavalli and C. Buonavoglia, “A Minor Groove Binder Probe Real-Time PCR Assay for Discrimination Between Type 2-Based Vaccines and Field Strains of Canine Parvovirus,” Journal of Virological Methods, Vol. 136, No. 1-2, 2006, pp. 65-70. doi:10.1016/j.jviromet.2006.03.030
[16]	P. Sagazio, M. Tempesta, D. Buonavoglia, F. Cirone and C. Buonavoglia, “Antigenic Characterization of Canine Parvovirus Strains Isolated in Italy,” Journal of Virological Methods, Vol. 73, No. 2, 1998, pp. 197-200. doi:10.1016/S0166-0934(98)00055-X
[17]	A. Steinel, E. H. Venter, M. Van Vuuren, C. R. Parrish and U. Truyen, “Antigenic and Genetic Analysis of Canine Parvoviruses in Southern Africa,” Onderstepoort Journal of Veterinary Research, Vol. 65, No. 4, 1998, pp. 239-242.
[18]	D. Buonavoglia, A. Cavalli, A. Pratelli, V. Martella, G. Greco M. Tempesta and C. Buonavoglia, “Antigenic Analysis of Canine Parvovirus Strains Isolated in Italy,” New Microbiology, Vol. 23, No. 1, 2000, pp. 93-96.
[19]	U. Truyen, A. Steinel, L. Bruckner, H. Lutz and K. Mostl, “Distribution of Antigen Types of Canine Parvovirus in Switzerland,” Austria and Germany. Schweiz Arch Tierheilkd, Vol. 142, No. 3, 2000, pp. 115-119.
[20]	V. Martella, A. Cavalli, A. Pratelli, G. Bozzo, M. Camero, D. Buonavoglia, D. Narcisi, M. Tempesta and C. Buonavoglia, “A Canine Parvovirus Mutant Is Spreading in Italy,” Journal of Clinical Microbiology, Vol. 42, No. 3, 2004, pp. 1333-1336. doi:10.1128/JCM.42.3.1333-1336.2004
[21]	N. Decaro, C. Desario, D. D. Addie, V. Martella, M. J. Vieira, G. Elia, A. Zicola, C. Davis, G. Thompson, E. Thiry, U. Truyen and C. Buonavoglia, “The Study Molecular Epidemiology of Canine Parvovirus”, Emerging Infectous Diseases, Vol. 13, No. 8, 2007, pp. 1222-1224. doi:10.3201/eid1308.070505
[22]	C. Hong, N. Decaro, C. Desario, P. Tanner, M. C. Pardo, S. Sanchez, C. Buonavoglia and J. T. Saliki, “Occurrence of Canine Parvovirus Type 2c in the United States,” Journal of Veterinary Diagnostic Investigation, Vol. 19, No. 5, 2007, pp. 535-539. doi:10.1177/104063870701900512
[23]	R. Perez, L. Francia, V. Romero, L. Maya, I. Lopez and M. Hernandez, “First Detection of Canine Parvovirus Type 2c in South America,” Veterinary Microbiology, Vol. 124, No. 1-2, 2007, pp. 147-152. doi:10.1016/j.vetmic.2007.04.028
[24]	N. Spibey, N. M. Greenwood, D. Sutton, W. S. K. Chalmers and I. Tarpey, “Canine Parvovirus Type 2 Vaccine Protects Against Virulent Challenge with Type 2c Virus,” Veterinary Microbiology, Vol. 128, No. 1-2, 2008, pp. 48-55. doi:10.1016/j.vetmic.2007.09.015
[25]	D. Mottershead, I. van der Linden, C. H. von Bonsdorff, K. K. Keinanen and C. Oker-Blom, “Baculoviral Display of the Green Fluorescent Protein and Rubella Virus Envelope Proteins,” Biochemical and Biophysical Research Communications, Vol. 238, No. 3, 1997, pp. 717-722. doi:10.1006/bbrc.1997.7372
[26]	M. M. Rahman, M. S. Shalia and K. P. Gopinathan, “Baculovirus Display of Fusion Protein of Peste des Petits Ruminants Virus and Hemagglutination Protein of Rinderpest Virus and Immunogenicity of the Displayed Proteins in Mouse Model,” Virology, Vol. 317, No. 1, 2003, pp. 36-49. doi:10.1016/j.virol.2003.08.022
[27]	X. G. Xu and H. J. Liu, “Baculovirus Surface Display of E2 Envelope Glycoprotein of Classical Swine Fever Virus and Immunogenicity of the Displayed Proteins in a Mouse Model,” Vaccine, Vol. 26, No. 43, 2008, pp. 5455-5460.
[28]	X. G. Xu, D. W. Tong, M. T. Chiou, Y. C. Hsieh, W. L. Shih, C. D. Chang, M. H. Liao, Y. M. Zhang, H. J. Liu, “Baculovirus Surface Display of NS3 Nonstructural Protein of Classical Swine Fever Virus,” Journal of Virological Methods, Vol.159, No. 2, 2009, pp. 259-264. doi:10.1016/j.jviromet.2009.04.013
[29]	X. G. Xu, Z. S. Wang, G. Zhang, Z. C. Li, H. N. Zhao, W. Li, D. W. Tong and H. J. Liu, “Baculovirus Surface Display of E Envelope Glycoprotein of Japanese Encephalitis Virus and Its Immunogenicity of the Displayed Proteins in Mouse and Swine Models,” Vaccine, Vol. 29, No. 4, 2011, pp. 636-643. doi:10.1016/j.vaccine.2010.11.045
[30]	X. G. Xu, Z. S. Wang, G. Zhang, Z. C. Li, H. N. Zhao, W. Li, D. W. Tong and H. J. Liu, “Baculovirus Virions Displaying Infectious Bursal Disease Virus VP2 Protein Protect Chickens Against Infectious Bursal Disease Virus Infection,” Avian Diseases, Vol. 55, No. 2, 2011, pp. 223-229. doi:10.1637/9597-111210-Reg.1
[31]	A. G. Oomens, S. A. Monsma and G. W. Blissard, “The Baculovirus GP64 Envelope Fusion Protein: Synthesis, Oligomerization and Processing,” Virology, Vol. 209, No. 2, 1995, pp. 592-603. doi:10.1006/viro.1995.1291