School of Life Sciences, Zhengzhou University, Zhengzhou 450001, China.
Key Laboratory of Animal Immunology of the Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Int J Biol Macromol. 2021 Jul 31;183:1393-1401. doi: 10.1016/j.ijbiomac.2021.05.053. Epub 2021 May 11.
Bluetongue (BT) is a non-contact infectious disease caused by Bluetongue virus (BTV), which can be transmitted by vector insects such as Culicoides and Aedes mosquitoes. The BTV VP2 protein encoded by the L2 gene is located at the outermost layer of the virus particle, plays a key role on mediating the adsorption and entry of virus, and it is also a main antigenic protein widely used for vaccine development. In this study, the BTV1 VP2 gene was cloned into pFastBac™Dual vector, and expressed in insect Sf21 cells. Immunized mice with purified recombinant VP2 protein can induce higher levels of antibodies. Three anti BTV1 VP2 monoclonal antibodies (mAbs) were generated (17E9C6, 17E9C8, 17E9H12), and showed high specific reactivity with recombinant VP2 protein and inactivated BTV1 virus. Finally, a novel linear B-cell epitope KEPAD on recombinant VP2 protein was identified by using three mAbs react with a series of continue-truncated peptides. The results of this study may provide new information on the structure and function of BTV1 VP2 protein and lay a foundation for the development of BTV1 diagnostic and prophylactic methods.
蓝舌病（BT）是一种由蓝舌病毒（BTV）引起的非接触性传染病，可通过媒介昆虫如库蠓和伊蚊传播。由 L2 基因编码的 BTV VP2 蛋白位于病毒粒子的最外层，在介导病毒的吸附和进入中起关键作用，也是广泛用于疫苗开发的主要抗原蛋白。本研究将 BTV1 VP2 基因克隆到 pFastBac™Dual 载体中，并在昆虫 Sf21 细胞中表达。用纯化的重组 VP2 蛋白免疫的小鼠可诱导更高水平的抗体。生成了三种抗 BTV1 VP2 的单克隆抗体（mAbs）（17E9C6、17E9C8、17E9H12），它们与重组 VP2 蛋白和灭活的 BTV1 病毒具有高度特异性反应。最后，通过三种 mAbs 与一系列连续截断肽的反应，鉴定了重组 VP2 蛋白上新的线性 B 细胞表位 KEPAD。本研究的结果可能为 BTV1 VP2 蛋白的结构和功能提供新信息，并为 BTV1 诊断和预防方法的开发奠定基础。
