[Construction and immunogenicity of a recombinant adenovirus co-expressing the E2 protein of classical swine fever virus and the porcine interleukin 2 in rabbits].
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.
College of Veterinary Medicine, Northwest A & F University, Yangling 712100, China. he-lei4280546@163.com
Lei
L
He
Yan-ming
YM
Zhang
Yan-zhao
YZ
Xu
Qing-hai
QH
Tang
Jing
J
Wang
Xiao-yun
XY
Yang
Chen
C
Dai
Hua
H
Xiang
Peng-xiang
PX
Chang
Zhi
Z
Lin
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D006692:Classical Swine Fever Virus / Q000235:genetics; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D013552:Swine; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D014765:Viral Vaccines / Q000276:immunology
D007376:Interleukin-2; D014764:Viral Proteins; D014765:Viral Vaccines