Li Zhipeng, Cui Kuiqing, Huang Kongwei, Liu Fuhang, Shi Deshun, Liu Qingyou
State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, Guangxi, 530005, China.
Agricultural Genomics Institute, Chinese Academy of Agricultural Sciences, Shenzhen, Guangdong, 518120, China.
Protein Pept Lett. 2019;26(12):904-909. doi: 10.2174/0929866526666190820161328.
Rotavirus is the most common cause of infectious diarrhea in infants and young children around the world. The inner capsid protein VP6 has been discussed as alternative vaccine as it can induce cross-protective immune responses against different RV strai. The use of ferritin nanoparticle may enhance the immunogenicity of the subunit vaccine.
In this article, our motivation is to design and obtain a self-assemble rotavirus nanoparticle vaccine which can induce efficiency immune response.
The VP6 protein was fused with ferritin and expressed in the Escherichia coli expression system. The recombinant VP6-ferritin (rVP6-ferritin) protein was purified by His-tag affinity chromatography and fast protein liquid chromatography. Transmission electron micrographic analysis was used to detect the nanostructure of the self-assembled protein. Mice were gavage with the protein and ELISA was used to detect the titer of the VP6 specific antibody.
The recombined VP6-ferritin was expressed in the Escherichia coli as an inclusion body form and the purified protein has similar antigenicity to rotavirus VP6. Transmission electron micrographic analysis of rVP6-ferritin exhibited spherical architecture with a uniform size distribution, which is similar to the ferritin nanocage. Immune response analysis showed that mice immunized by rVP6-ferritin protein induced 8000 (8000±1093) anti-VP6 IgG titers or 1152 (1152±248.8) anti-VP6 IgA titers.
According to the above research, the rotavirus VP6-ferritin protein can be easily express and self-assemble to the nano-vaccine and induce efficiency humoral and mucosal immunity. Our research makes a foundation for the development of oral rotavirus vaccine.
轮状病毒是全球婴幼儿感染性腹泻最常见的病因。内衣壳蛋白VP6可诱导针对不同轮状病毒毒株的交叉保护性免疫反应,因此被视为替代疫苗。使用铁蛋白纳米颗粒可能会增强亚单位疫苗的免疫原性。
本文旨在设计并获得一种能诱导高效免疫反应的自组装轮状病毒纳米颗粒疫苗。
将VP6蛋白与铁蛋白融合,在大肠杆菌表达系统中表达。通过组氨酸标签亲和色谱和快速蛋白质液相色谱法纯化重组VP6-铁蛋白(rVP6-铁蛋白)蛋白。采用透射电子显微镜分析检测自组装蛋白的纳米结构。给小鼠灌胃该蛋白,并用酶联免疫吸附测定法检测VP6特异性抗体的效价。
重组VP6-铁蛋白在大肠杆菌中以包涵体形式表达,纯化后的蛋白与轮状病毒VP6具有相似的抗原性。对rVP6-铁蛋白的透射电子显微镜分析显示其呈球形结构,大小分布均匀,与铁蛋白纳米笼相似。免疫反应分析表明,用rVP6-铁蛋白免疫的小鼠诱导出8000(8000±1093)的抗VP6 IgG效价或1152(1152±248.8)的抗VP6 IgA效价。
根据上述研究,轮状病毒VP6-铁蛋白可轻松表达并自组装成纳米疫苗,诱导高效的体液免疫和黏膜免疫。本研究为口服轮状病毒疫苗的开发奠定了基础。