柯萨奇病毒A组16型病毒样颗粒免疫原性的初步研究

目的 研究柯萨奇病毒A组16型（Coxsackievirus A16,CA16）病毒样颗粒（Virus-like particles,VLPs）的免疫原性.方法 利用杆状病毒-昆虫细胞表达系统共表达3CD及P1蛋白,制备CA16病毒样颗粒,通过SDS-PAGE及透射电镜等方法对VLPs进行鉴定,以氢氧化铝佐剂免疫ICR小鼠,并对乳鼠经颅腔攻毒.对病毒样颗粒疫苗的免疫原性及保护效果进行评价.结果 将重组CA163CD及P1蛋白的杆状病毒转染SF9细胞,可以产生类似CA16病毒颗粒的大小为27 ～ 30 nm的VLPs.小鼠免疫试验结果显示CA16 VLPs可以刺激产生较高水平的抗CA16病毒的特异性IgG抗体及中和抗体,乳鼠攻毒试验结果显示,母传抗体保护率高达90％.结论 CA16 VLPs可以刺激小鼠产生较高水平的体液免疫反应,并且母传抗体可以保护乳鼠抵御经颅腔的病毒攻击.

Coxsackievirus A16 virus-like particles protect mice against lethal challenge

Objective To study the immunogenicity of virus-like particles （VLPs） of Coxsackievirus A16.Methods Recombinant CA16 VLPs were prepared by co-expressing 3CD and P1 proteins using baculovirus expression system.Purified VLPs were identified by SDS-PAGE and transmission electronmicroscope（TEM）.The immunogenicity of CA16 VLPs was evaluated in ICR mice.Humoral immune response was identified by ELISA and neutralization assay and the protection effects the neonatal mouse against CA16 virus were also evaluated.Results Recombinant CA16 VLPs exhibited as electrondense spherical particles of 27-30 nm size,which were similar to the morphology and size of authentic virions under TEM observation.The VLPs induced high levels of CA16 virus-specific serum IgG antibodies and neutralization antibodies.Furthermore,VLPs-immunized mother mice conferred protection （survival rate up to 90%） on neonatal mice against a lethal CA16 challenge of 104 plaque-forming units CA16.Conclusion CA16 VLPs produced by baculovirus expression system were capable to induce specific humoralimmune responses in mice and the neutralization antibodies could protect the neonatal mouse from lethal CA16 virus challenge.