猪轮状病毒VP8截短蛋白-铁蛋白纳米颗粒的制备与免疫原性分析  

Preparation and Immunogenicity Analysis of VP8 Truncated Protein-Ferritin Nanoparticles of Porcine Rotavirus

作　　者：彭凤巧 李博硕 杜恩岐[1] 周宏超[1] PENG Feng-qiao;LI Bo-shuo;DU En-qi;ZHOU Hong-chao(College of Veterinary Medicine,Northwest A&F University,Yangling,Shaanxi 712100,China;Yangling Kare Biotechnology Co.,LTD.,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学动物医学院,陕西杨陵712100 [2]杨凌凯瑞生物科技有限公司,陕西杨陵712100

出　　处：《动物医学进展》2025年第4期59-65,共7页Progress In Veterinary Medicine

基　　金：陕西省农业厅省级农业专项(2019SJNYZX45)。

摘　　要：在铁蛋白纳米颗粒骨架上集中展示猪轮状病毒(PoRV)VP8截短蛋白,制备一种以铁蛋白(Ferritin)为载体的PoRV纳米颗粒亚单位疫苗并评估其免疫原性。将PoRV VP8截短蛋白和铁蛋白利用SpyCatcher/SpyTag系统连接形成PoRV VP8-Ferritin,通过透射电镜(TEM)、高效液相色谱(HPLC)、动态光散射(DLS)对纳米颗粒进行表征分析,并配伍佐剂制备亚单位疫苗免疫小鼠。对免疫后各时间点小鼠血清的特异性IgG抗体和35 d的特异性IgG抗体进行分型(IgG2a和IgG1)检测。结果显示,VP8-Ferritin能较好地在体外组装形成平均粒径为15.56 nm的颗粒结构,组装效率可达85%。免疫后14 d VP8-Ferritin免疫组和VP8免疫组均能在小鼠血清中检测到特异性IgG抗体,OD 450值分别为0.521和0.229;免疫后42 d各免疫组的特异性IgG抗体水平达到峰值,OD 450值分别为1.197和0.531。免疫后的VP8-Ferritin亚单位疫苗能够诱导小鼠产生较高水平的特异性IgG抗体,其特异性IgG抗体水平极显著高于VP8亚单位疫苗(P<0.01);VP8-Ferritin免疫组IgG2a/IgG1比值为2.564,显著高于VP8免疫组的IgG2a/IgG1比值1.791(P<0.05)。结果表明VP8-Ferritin纳米颗粒亚单位疫苗增强了VP8抗原的免疫原性,主要表现TH1型免疫反应,该结果为猪轮状病毒亚单位疫苗的研究提供了新思路。Porcine rotavirus(PoRV)VP8 truncated protein was exhibited on the skeleton of ferritin nanoparticles to prepare a PoRV nanoparticle subunit vaccine with ferritin as carrier and evaluate its immunogenicity.PoRV VP8 truncated protein and ferritin were connected by SpyCatcher/SpyTag system to form PoRV VP8-Ferritin,and the nanoparticles were characterized by transmission electron microscopy(TEM),high performance liquid chromatography(HPLC)and dynamic light scattering(DLS).Subunit vaccine was prepared with adjuvant to immunize mice.The specific IgG antibodies of mice at different time points after immunization and specific IgG antibodies at 35 days after immunization were classified(IgG2a and IgG1).The results showed that VP8-Ferritin can be assembled in vitro with an average particle size of 15.56 nm,and the assembly efficiency can reach 85%.14 days after immunization,specific IgG antibody was detected in serum in both VP8-Ferritin and VP8 groups,with OD 450 values of 0.521 and 0.229,respectively.The specific IgG antibody levels reached the peak at day 42 after immunization,with OD 450 levels of 1.197 and 0.531,respectively.After immunization,VP8-Ferritin subunit vaccine could induce mice to produce higher level of specific IgG antibody,and the level of specific IgG antibody was significantly better than that of VP8 subunit vaccine(P<0.01).The IgG2a/IgG1 ratio in VP8-Ferritin group was 2.564,which was significantly higher than the IgG2a/IgG1 ratio of 1.791 in VP8 group(P<0.05).The results showed that VP8-Ferritin nanoparticle subunit vaccine enhanced the immunogenicity of VP8 antigen,mainly manifested TH1 immune response,which provided a new idea for the study of porcine rotavirus subunit vaccine.

关 键 词：猪轮状病毒 VP8蛋白 纳米颗粒 免疫原性 

分 类 号：S852.4[农业科学—基础兽医学] S858.28[农业科学—兽医学]