绵羊痘P32蛋白嵌合型病毒样颗粒的构建及免疫原性评价

Construction and immunogenicity evaluation of chimeric virus-like particles of sheep pox P32 protein

基于病毒样颗粒生产平台,以绵羊痘病毒P32蛋白为研究靶点,通过GPI锚定策略将P32蛋白锚定至新城疫病毒样颗粒囊膜表面制备嵌合型病毒样颗粒cVLP-P32。透射电镜可见近圆形的病毒粒子上布满纤突,Western blot结果显示各组分蛋白均正确表达。随后以BALB/c小鼠和绵羊为试验动物,评价cVLP-P32的免疫原性,ELISA结果显示cVLP-P32能够诱导小鼠及绵羊产生较高滴度的特异性抗体;流式细胞术分析结果显示,与可溶性GPI-P32蛋白相比,cVLP-P32可诱导小鼠激发更强的CD3^+CD4^+T及CD3^+CD8^+T细胞免疫应答。

In this research,the Bac to Bac platform was used to produce a sheep pox virus P32 protein decorated Newcastle disease virus-like particle(NDV VLPs).The P32 protein of sheep pox virus was anchored to the surface of NDV VLPs membrane through a GPI mediated strategy to prepare the chimeric virus-like particles(cVLPs-P32).Transmission electron microscopy(TEM)showed that the cVLPs-P32 were covered with fibrils.Western blot results showed that the proteins of each component were correctly expressed.Subsequently,BALB/c mice and sheep were used as experimental animals to evaluate the immunogenicity of cVLPs-P32.ELISA results showed that cVLPs-P32 could induce induce higher titer of specific antibodies in mice and sheep.Flow cytometry analysis showed that cVLPs-P32 could stronger cellular immune response than soluble GPI-P32 protein in mice,indicated by higher percentage of splenic CD3^+CD4^+T and CD3^+CD8^+T cells.