摘要
White spot syndrome virus (WSSV) is a large dsDNA virus responsible for white spot disease in shrimp and other crustaceans. VP28 is one of the major envelope proteins of WSSV and plays a crucial role in viral infection. In an effort to develop a vaccine against WSSV, we have constructed a recombinant baculovirus with an immediate early promoter 1 which expresses VP28 at an early stage of infection in insect cells. Baculovirus expressed rVP28 was able to maintain its structural and antigenic conformity as indicated by immunofluorescence assay and western blot analysis. Interestingly, our results with confocal microscopy revealed that rVP28 was able to localize on the plasma membrane of insect cells infected with recombinant baculovirus. In addition, we demonstrated with transmission electron microscopy that baculovirus successfully acquired rVP28 from the insect cell membrane via the budding process. Using this baculovirus displaying VP28 as a vaccine against WSSV, we observed a significantly higher survival rate of 86.3% and 73.5% of WSSV-infected shrimp at 3 and 15 days post vaccination respectively. Quantitative real-time PCR also indicated that the WSSV viral load in vaccinated shrimp was significantly reduced at 7 days post challenge. Furthermore, our RT-PCR and immunohistochemistry results demonstrated that the recombinant baculovirus was able to express VP28 in vivo in shrimp tissues. This study will be of considerable significance in elucidating the morphogenesis of WSSV and will pave the way for new generation vaccines against WSSV.
摘要译文
白点综合征病毒（WSSV）是一种负责虾和其他甲壳类动物白斑病的大型dsDNA病毒VP28是WSSV的主要包膜蛋白之一，在病毒感染中起关键作用。为了开发针对WSSV的疫苗，我们构建了一种具有立即早期启动子1的重组杆状病毒，其在昆虫细胞的感染早期表达VP28表达rVP28的杆状病毒能够通过免疫荧光测定和蛋白质印迹分析来保持其结构和抗原依从性。有趣的是，我们的共聚焦显微镜检查结果显示，rVP28能够定位在重组杆状病毒感染的昆虫细胞的质膜上。另外，我们用透射电子显微镜观察到杆状病毒通过发芽过程从昆虫细胞膜成功获得rVP28使用显示VP28的杆状病毒作为针对WSSV的疫苗，我们观察到显着更高的存活率为86 5和73 疫苗接种后3和15天，WSSV感染虾5％25定量实时荧光定量PCR也表明，攻击后7天，疫苗接种的虾的WSSV病毒载量显着降低。此外，我们的RT-PCR和免疫组织化学结果表明，重组杆状病毒能够在虾组织中体内表达VP28本研究对阐明WSSV形态发生具有重要意义，为新一代针对WSSV疫苗的疫苗铺路
S. Syed Musthaq[a]; Selvaraj Madhan[a]; A.S. Sahul Hameed[b]; Jimmy Kwang[a][c]. Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon[J]. Virology, 2009,391(2): 315–324