[Construction and immunogenicity of recombinant adenovirus co-expressing the GP5 and M protein of porcine reproduction and respriratory syndrome virus in mice].
FMDV 2A peptide was introduced as a linker between GP5 and M protein of porcine reproduction and respiratory syndrome virus (PRRSV) to allow automatic self-cleavage the polyproteins. This strategy simultaneously displayed the neutralizing action of GP5 protein and cell-mediated immunity of M protein. We put them into the expression cassette of adenovirus vector. The results of RT-PCR, IFA and Western blotting showed that GP5 and M protein were not only expressed correctly, but also self-cleavaged and assemble heterodimers formation. To detect the advantages of rAd-GP5-2A-M, we also constructed some other recombinant adenoviruses (rAd-GP5, rAd-M and rAd-GP5-M) as control. After inoculated subcutaneously into BALB/c mice, the four recombinant adenoviruses can induce PRRSV-specific antibodies and cell-mediated immune response, but the level of humoral and cell-mediated immune response against PRRSV induced by rAd-GP5-2A-M is the strongest among the four recombinant adenoviruses. All of these suggested that it is possible to develop one multi-gene engineering vaccine utilizing FMDV 2A peptide, and also provided a novel strategy for developing other viral disease vaccine.
Yun
Tao
T
Research Institute of Virology & Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
Ni
Zheng
Z
Yu
Bin
B
Chen
Liu
L
Hua
Jionggang
J
Wang
Genrong
G
Liu
Guangqing
G
D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D005260:Female; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D013552:Swine; D014614:Vaccines, Synthetic / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014765:Viral Vaccines / Q000276:immunology*
C525999:M protein, porcine reproductive and respiratory syndrome virus; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; D014765:Viral Vaccines; C114789:glycoprotein 5, PRRSV