Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China; Key Laboratory of Animal Diseases Diagnostic and Immunology, College of Veterinary Medicine of Nanjing Agricultural University, Nanjing 210095, China.
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
Virus Res. 2014 Apr;183:15-22. doi: 10.1016/j.virusres.2014.01.004. Epub 2014 Jan 20.
To explore the capacity and immunogenicity of virus-like particles (VLPs) of rabbit hemorrhagic disease virus (RHDV) accommodating foreign amino acid sequences, integrations were performed at the following four locations of the structural protein VP60 of RHDV using the OVA257-264 CD8+ T cell epitope (SIINFEKL): (1) inserting at the N-terminus of the VP60 protein (N1); (2) replacing amino acid positions 2-14 of the VP60 protein (N2); (3) replacing amino acid positions 196-207 of the VP60 protein (I1); and (4) replacing amino acid positions 217-228 of the VP60 protein (I2). The recombinant proteins were expressed by baculovirus expression system. The ability to form RHDV-like particles was confirmed by electron microscopy. The immunogenicity of the four recombinant proteins (N1, N2, I1 and I2) was evaluated in mice without any adjuvants. The results indicated that the four recombinant proteins (N1, N2, I1 and I2) could assemble into VLPs. All of the recombinant proteins could induce a specific immune response. Recombinant proteins I1 and I2 were able to elicit both high levels of IFN-γ secretion and anti-VP60 specific immune responses in the murine model. The levels of the VP60-specific IgG antibody in groups I1 and I2 displayed higher optical density (OD) values than those of groups N1 and N2 (P<0.001, P<0.001). The number of IFN-γ-producing splenocytes in mice that were immunized with recombinant proteins I1 and I2 was also significantly greater compared with mice that were immunized with recombinant proteins N1 and N2 (P<0.01). All of these above mentioned results might be beneficial to the establishment of the RHDV-VLPs display system.
为了探索容纳外源氨基酸序列的兔出血症病毒（RHDV）病毒样颗粒（VLPs）的容量和免疫原性，利用 OVA257-264 CD8+ T 细胞表位（SIINFEKL），在 RHDV 结构蛋白 VP60 的以下四个位置进行了整合：（1）插入 VP60 蛋白的 N 端（N1）；（2）替换 VP60 蛋白的氨基酸位置 2-14（N2）；（3）替换 VP60 蛋白的氨基酸位置 196-207（I1）；和（4）替换 VP60 蛋白的氨基酸位置 217-228（I2）。重组蛋白通过杆状病毒表达系统表达。通过电子显微镜确认了形成 RHDV 样颗粒的能力。在没有任何佐剂的情况下，在小鼠中评估了四种重组蛋白（N1、N2、I1 和 I2）的免疫原性。结果表明，四种重组蛋白（N1、N2、I1 和 I2）能够组装成 VLPs。所有重组蛋白均能诱导特异性免疫反应。重组蛋白 I1 和 I2 能够在小鼠模型中引起高水平的 IFN-γ 分泌和抗 VP60 特异性免疫反应。与 N1 和 N2 组相比，I1 和 I2 组的 VP60 特异性 IgG 抗体的 OD 值更高（P<0.001，P<0.001）。与 N1 和 N2 组相比，免疫重组蛋白 I1 和 I2 的小鼠脾细胞中 IFN-γ 产生细胞的数量也显著增加（P<0.01）。所有这些结果都可能有助于建立 RHDV-VLPs 展示系统。
Zotero 插件