Orf virus DNA vaccines expressing ORFV 011 and ORFV 059 chimeric protein enhances immunogenicity.
BACKGROUND
ORFV attenuated live vaccines have been the main prophylactic measure against contagious ecthyma in sheep and goats in the last decades, which play an important role in preventing the outbreak of the disease. However, the available vaccines do not induce lasting immunity in sheep and goats. On the other hand, variation in the terminal genome of Orf virus vaccine strains during cell culture adaptation may affect the efficacy of a vaccine. Currently, there are no more effective antiviral treatments available for contagious ecthyma.


RESULTS
We constructed three eukaryotic expression vectors pcDNA3.1-ORFV011, pcDNA3.1-ORFV059 and pcDNA3.1-ORFV011/ORFV059 and tested their immunogenicity in mouse model. High level expression of the recombinant proteins ORFV011, ORFV059 and ORFV011/ORFV059 was confirmed by western blotting analysis and indirect fluorescence antibody (IFA) tests. The ORFV-specific antibody titers and serum IgG1/IgG2a titers, the proliferation of lymphocytes and ORFV-specific cytokines (IL-2, IL-4, IL-6, IFN-γ, and TNF-α) were examined to evaluate the immune responses of the vaccinated mice. We found that mice inoculated with pcDNA3.1-ORFV 011/ORFV059 had significantly stronger immunological responses than those inoculated with pcDNA3.1-ORFV011, pcDNA3.1-ORFV059, or pcDNA3.1-ORFV011 plus pcDNA3.1-ORFV059. Compared to other vaccine plasmids immunized groups, pcDNA3.1-ORFV011/ORFV059 immunized group enhances immunogenicity.


CONCLUSIONS
We concluded that DNA vaccine pcDNA3.1-ORFV011/ORFV059 expressing ORFV011 and ORFV059 chemeric-proteins can significantly improve the potency of DNA vaccination and could be served as more effective and safe approach for new vaccines against ORFV.
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China.
Kui
K
Zhao
Wenqi
W
He
Wei
W
Gao
Huijun
H
Lu
Tiesuo
T
Han
Jing
J
Li
Ximu
X
Zhang
Bingbing
B
Zhang
Gaili
G
Wang
Gaoli
G
Su
Zhihui
Z
Zhao
Deguang
D
Song
Feng
F
Gao
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology; D015153:Blotting, Western; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology; D004474:Ecthyma, Contagious / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006041:Goats; D051379:Mice; D008807:Mice, Inbred BALB C; D009923:Orf virus / Q000235:genetics* / Q000276:immunology; D010957:Plasmids; D061251:Primary Cell Culture; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology; D012756:Sheep; D014162:Transfection; D014420:Turbinates / Q000166:cytology / Q000378:metabolism*; D014611:Vaccination; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D016207:Cytokines; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D014764:Viral Proteins; D014765:Viral Vaccines