School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215123, China.
Fish Shellfish Immunol. 2013 Jan;34(1):348-55. doi: 10.1016/j.fsi.2012.11.024. Epub 2012 Nov 29.
To obtain a vaccine and evaluate its immune efficacy as an oral vaccine against Grass Carp Reovirus (GCRV), a donor vector pFastBac-FA-VP6-ph-VP6 containing two GCRV vp6 genes, one driven by the Megalobrama amblycephala β-actin promoter and another one controlled by baculovirus polyhedrin promoter was constructed to generate the recombinant baculovirus BacFish-vp6. From the hemolymph of 5th instar silkworm inoculated with BacFish-vp6, a 53 kDa recombinant VP6 protein could be detected. And the infected pupae collected at 120 h post-inoculation with BacFish-vp6 were used to make freeze-dried powder as an oral vaccine. When the grass carps were orally administrated with feed containing 1%, 5% and 10% of the freeze-dried powder, their specific antibody against VP6 could be detected. Further studies showed that vp6 has been transcripted in the grass carp kidney (CIK) cells infected with BacFish-vp6 and the different tissues of orally vaccinated fish. All the results suggested the powder of the silkworm pupae infected with BacFish-vp6 could possibly be used as an orally administered vaccine.
为了获得一种疫苗,并评估其作为草鱼呼肠孤病毒(GCRV)口服疫苗的免疫效果,构建了一个携带两个 GCRV vp6 基因的供体载体 pFastBac-FA-VP6-ph-VP6,其中一个由鳊鱼 β-肌动蛋白启动子驱动,另一个由杆状病毒多角体蛋白启动子控制,以生成重组杆状病毒 BacFish-vp6。从接种 BacFish-vp6 的五龄期家蚕的血淋巴中,可以检测到 53 kDa 的重组 VP6 蛋白。并且在接种 BacFish-vp6 后 120 小时收集的感染蛹被用来制备冻干粉末作为口服疫苗。当草鱼口服含有 1%、5%和 10%冻干粉末的饲料时,可以检测到针对 VP6 的特异性抗体。进一步的研究表明,vp6 在感染 BacFish-vp6 的草鱼肾脏(CIK)细胞和口服疫苗鱼的不同组织中被转录。所有结果表明,感染 BacFish-vp6 的蚕蛹粉末可能可用作口服疫苗。