Construction and immunogenicity of a recombinant adenovirus co-expressing the E2 protein of classical swine fever virus and the porcine interleukin 2 in rabbits.
- Author:
Lei HE
1
;
Yan-ming ZHANG
;
Yan-zhao XU
;
Qing-hai TANG
;
Jing WANG
;
Xiao-yun YANG
;
Chen DAI
;
Hua XIANG
;
Peng-xiang CHANG
;
Zhi LIN
Author Information
1. College of Veterinary Medicine, Northwest A & F University, Yangling 712100, China. he-lei4280546@163.com
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
metabolism;
Animals;
Cell Line;
Classical swine fever virus;
genetics;
Humans;
Interleukin-2;
genetics;
immunology;
Rabbits;
Swine;
Viral Proteins;
genetics;
immunology;
metabolism;
Viral Vaccines;
immunology
- From:
Chinese Journal of Virology
2010;26(5):385-391
- CountryChina
- Language:Chinese
-
Abstract:
To construct a recombinant adenovirus co-expressing the E2 protein of classical swine fever virus (CSFV) and the porcine interleukin 2 (pIL-2), the CSFV E2 gene and pIL-2 gene were amplified respectively from the plasmids pMD19-T-E2 and pMD19-T-pIL-2 by PCR. E2-pIL-2 fusion gene was obtained by using 5 consecutive glycine codons as a linker and cloned into the adenoviral shuttle plasmid AdTrack. The AdTrack-E2-pIL-2 was linearized and transformed into E. coli BJ5183 with the backbone plasmid AdEasy1. The resultant recombinant plasmid AdEasy-E2-pIL-2 was transfected into the 293 cells where the recombinant adenovirus rAd-E2-pIL-2 was produced. The immunogenicity of rAd-E2-pIL-2 was evaluated in rabbits. The results of RT-PCR and Western-blotting showed that rAd-E2-pIL-2 could carry and express E2 and pIL-2 proteins. The titer of the rAd-E2-pIL-2 was 10(8.12) PFU/mL. After immunized with rAd-E2pIL-2, The injected rabbits developed a high level of CSFV specific antibodies. Regular fever was not detected in the rAd-E2-pIL-2-immunized rabbits upon challenge with CSFV C stain, and specific lymphoproliferative responses to the CSFV was detected in the lymphocytes from the immunized rabbits. In conclusion, rAd-E2-pIL-2 was constructed successfully and it could be an attractive vaccine candidate against CSFV.
