Yun Tao, Ni Zheng, Yu Bin, Chen Liu, Hua Jionggang, Wang Genrong, Liu Guangqing
Research Institute of Virology & Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
Sheng Wu Gong Cheng Xue Bao. 2009 Apr;25(4):488-95.
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.
口蹄疫病毒2A肽被引入作为猪繁殖与呼吸综合征病毒(PRRSV)的GP5和M蛋白之间的连接子,以使多聚蛋白能够自动自我切割。该策略同时展示了GP5蛋白的中和作用和M蛋白的细胞介导免疫。我们将它们放入腺病毒载体的表达盒中。RT-PCR、IFA和Western印迹结果表明,GP5和M蛋白不仅正确表达,而且能自我切割并组装形成异源二聚体。为了检测rAd-GP5-2A-M的优势,我们还构建了一些其他重组腺病毒(rAd-GP5、rAd-M和rAd-GP5-M)作为对照。皮下接种到BALB/c小鼠后,这四种重组腺病毒均可诱导PRRSV特异性抗体和细胞介导的免疫反应,但rAd-GP5-2A-M诱导的针对PRRSV的体液免疫和细胞介导免疫反应水平在这四种重组腺病毒中最强。所有这些表明,利用口蹄疫病毒2A肽开发一种多基因工程疫苗是有可能的,也为开发其他病毒性疾病疫苗提供了一种新策略。