Department of Biotechnology Pukyong National University, Busan, 608-737, Korea.
J Microbiol Biotechnol. 2011 Feb;21(2):170-5. doi: 10.4014/jmb.1005.05036.
This study was carried out for the molecular level identification of recombinant protein vaccine efficacy, by oral feeding against white spot syndrome virus infection, with the comparison of viral mRNA transcriptional levels in shrimp cells. For the determination of WSSV dilution ratio for the vaccination experiment by oral feeding, in vivo virus titration was carried out using different virus dilutions of virus stock (1×10(2), 2×10(2), and 1×10(3)). Among the dilution ratios, 2×10(2) diluted WSSV stock was chosen as the optimal condition because this dilution showed 90% mortality at 10 days after virus injection. Recombinant viral proteins, rVP19 and rVP28, produced as protein vaccines were delivered in shrimps by oral feeding. The cumulative mortalities of the shrimps vaccinated with rVP19 and rVP28 at 21 days after the challenge with WSSV were 66.7% and 41.7%, respectively. This indicates that rVP28 showed a better protective effect against WSSV in shrimp than rVP19. Through the comparison of mRNA transcriptional levels of viral genes from collected shrimp organ samples, it was confirmed that viral gene transcriptions of vaccinated shrimps were delayed for 4~10 days compared with those of unvaccinated shrimps. Protection from WSSV infection in shrimp by the vaccination with recombinant viral proteins could be accomplished by the prevention of entry of WSSV due to the shrimp immune system activated by recombinant protein vaccines.
本研究旨在通过口服途径对重组蛋白疫苗的疗效进行分子水平鉴定,以评估其对白斑综合征病毒(WSSV)感染的保护效果,并比较虾细胞中病毒 mRNA 转录水平的变化。为了确定口服途径接种疫苗实验中 WSSV 的稀释比例,我们进行了体内病毒滴定实验,使用不同稀释度的病毒储备液(1×10(2)、2×10(2)和 1×10(3))。在这些稀释比例中,选择了 2×10(2)稀释的 WSSV 储备液作为最佳条件,因为该稀释度在病毒注射后 10 天内可导致 90%的死亡率。作为蛋白疫苗的重组病毒蛋白 rVP19 和 rVP28 通过口服途径递送至虾体内。在接受 WSSV 攻毒后 21 天,rVP19 和 rVP28 疫苗接种虾的累积死亡率分别为 66.7%和 41.7%。这表明 rVP28 对虾的 WSSV 保护效果优于 rVP19。通过比较从收集的虾器官样本中病毒基因的 mRNA 转录水平,我们证实与未接种疫苗的虾相比,接种疫苗的虾的病毒基因转录延迟了 4-10 天。通过重组病毒蛋白疫苗接种激活虾的免疫系统,可以防止 WSSV 的进入,从而实现对虾 WSSV 感染的保护。