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Immunogenicity of a recombinant pseudorabies virus expressing ORF1-ORF2 fusion protein of porcine circovirus type 2.
Vet Microbiol. 2005 Aug 30; 109(3-4):179-90.VM

Abstract

Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS). Pseudorabies (PR) is also an important infectious disease in swine and sometimes co-infect with PCV2. An attenuated pseudorabies virus (PRV) has been successfully used as a vector for live viral vaccines. In this study, a recombinant PRV expressing ORF1-ORF2 fusion protein of PCV2 was constructed and its immunogenicity was tested in mice and pigs. The ORF1 and partial ORF2 gene of PCV2 Yu-A strain were amplified by PCR and inserted into a transfer vector. The recombinant transfer plasmid was co-transfected with the EcoRI digested genome of vector virus (PRV TK-/gE-/LacZ+) into IBRS-2 cells. The recombinant pseudorabies virus PRV-PCV2 was purified by plaque purification and identified by PCR and Southern blotting. Expression of the ORF1-ORF2 fusion protein by the recombinant PRV-PCV2 virus was demonstrated by Western blotting analysis. The growth properties of the recombinant virus in cells were similar to that of the parent vector virus. In animal experiments, PRV-PCV2 elicited strong anti-PRV and anti-PCV2 antibodies in Balb/c mice as indicated by PRV-neutralizing assay, anti-PCV2 ELISA and PCV2 specific lymphocyte proliferation assay, respectively. And PRV-PCV2 immunization protected mice against a lethal challenge of a virulent PRV Ea strain. In pigs, PRV-PCV2 elicited significant immune response towards PRV and PCV2 as indicated by PRV-ELISA, PRV neutralizing assay and PCV2 specific lymphocyte proliferation assay, respectively. This is a first step toward the development of a potential candidate divalent vaccine against PRV and PCV2 infections.

Authors+Show Affiliations

Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16006077

Citation

Ju, Chunmei, et al. "Immunogenicity of a Recombinant Pseudorabies Virus Expressing ORF1-ORF2 Fusion Protein of Porcine Circovirus Type 2." Veterinary Microbiology, vol. 109, no. 3-4, 2005, pp. 179-90.
Ju C, Fan H, Tan Y, et al. Immunogenicity of a recombinant pseudorabies virus expressing ORF1-ORF2 fusion protein of porcine circovirus type 2. Vet Microbiol. 2005;109(3-4):179-90.
Ju, C., Fan, H., Tan, Y., Liu, Z., Xi, X., Cao, S., Wu, B., & Chen, H. (2005). Immunogenicity of a recombinant pseudorabies virus expressing ORF1-ORF2 fusion protein of porcine circovirus type 2. Veterinary Microbiology, 109(3-4), 179-90.
Ju C, et al. Immunogenicity of a Recombinant Pseudorabies Virus Expressing ORF1-ORF2 Fusion Protein of Porcine Circovirus Type 2. Vet Microbiol. 2005 Aug 30;109(3-4):179-90. PubMed PMID: 16006077.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Immunogenicity of a recombinant pseudorabies virus expressing ORF1-ORF2 fusion protein of porcine circovirus type 2. AU - Ju,Chunmei, AU - Fan,Huiying, AU - Tan,Yadi, AU - Liu,Zhengfei, AU - Xi,Xin, AU - Cao,Shengbo, AU - Wu,Bin, AU - Chen,Huanchun, PY - 2004/12/10/received PY - 2005/05/08/revised PY - 2005/06/02/accepted PY - 2005/7/12/pubmed PY - 2005/10/7/medline PY - 2005/7/12/entrez SP - 179 EP - 90 JF - Veterinary microbiology JO - Vet. Microbiol. VL - 109 IS - 3-4 N2 - Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS). Pseudorabies (PR) is also an important infectious disease in swine and sometimes co-infect with PCV2. An attenuated pseudorabies virus (PRV) has been successfully used as a vector for live viral vaccines. In this study, a recombinant PRV expressing ORF1-ORF2 fusion protein of PCV2 was constructed and its immunogenicity was tested in mice and pigs. The ORF1 and partial ORF2 gene of PCV2 Yu-A strain were amplified by PCR and inserted into a transfer vector. The recombinant transfer plasmid was co-transfected with the EcoRI digested genome of vector virus (PRV TK-/gE-/LacZ+) into IBRS-2 cells. The recombinant pseudorabies virus PRV-PCV2 was purified by plaque purification and identified by PCR and Southern blotting. Expression of the ORF1-ORF2 fusion protein by the recombinant PRV-PCV2 virus was demonstrated by Western blotting analysis. The growth properties of the recombinant virus in cells were similar to that of the parent vector virus. In animal experiments, PRV-PCV2 elicited strong anti-PRV and anti-PCV2 antibodies in Balb/c mice as indicated by PRV-neutralizing assay, anti-PCV2 ELISA and PCV2 specific lymphocyte proliferation assay, respectively. And PRV-PCV2 immunization protected mice against a lethal challenge of a virulent PRV Ea strain. In pigs, PRV-PCV2 elicited significant immune response towards PRV and PCV2 as indicated by PRV-ELISA, PRV neutralizing assay and PCV2 specific lymphocyte proliferation assay, respectively. This is a first step toward the development of a potential candidate divalent vaccine against PRV and PCV2 infections. SN - 0378-1135 UR - https://www.unboundmedicine.com/medline/citation/16006077/Immunogenicity_of_a_recombinant_pseudorabies_virus_expressing_ORF1_ORF2_fusion_protein_of_porcine_circovirus_type_2_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0378-1135(05)00204-X DB - PRIME DP - Unbound Medicine ER -