Immunogenicity of a recombinant pseudorabies virus expressing ORF1-ORF2 fusion protein of porcine circovirus type 2.
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.
Ju
Chunmei
C
Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China.
Fan
Huiying
H
Tan
Yadi
Y
Liu
Zhengfei
Z
Xi
Xin
X
Cao
Shengbo
S
Wu
Bin
B
Chen
Huanchun
H
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D015153:Blotting, Western / Q000662:veterinary; D018173:Circoviridae Infections / Q000150:complications / Q000235:genetics / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary*; D017925:Circovirus / Q000235:genetics / Q000276:immunology*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000276:immunology*; D007114:Immunization / Q000662:veterinary; D051379:Mice; D008807:Mice, Inbred BALB C; D016366:Open Reading Frames; D011557:Pseudorabies / Q000150:complications / Q000235:genetics / Q000276:immunology* / Q000517:prevention & control; D011897:Random Allocation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013552:Swine; D013553:Swine Diseases / Q000276:immunology / Q000517:prevention & control / Q000821:virology*; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology
D000914:Antibodies, Viral; D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D014764:Viral Proteins; D014765:Viral Vaccines