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首页> 外文期刊>Veterinary Microbiology >Synthesis of empty capsid-like particles of Asia I foot-and-mouth disease virus in insect cells and their immunogenicity in guinea pigs
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Synthesis of empty capsid-like particles of Asia I foot-and-mouth disease virus in insect cells and their immunogenicity in guinea pigs

机译:昆虫细胞中亚洲I型口蹄疫病毒空衣壳样颗粒的合成及其在豚鼠中的免疫原性

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摘要

The assembly of foot-and-mouth disease virus (FMDV) requires the cleavage of the P12A polyprotein into individual structural proteins by protease 3C. In this study, we constructed a recombinant baculovirus that simultaneously expressed the genes for the P12A and 3C proteins of Asia I FMDV from individual promoters. The capsid proteins expressed in High Five insect cells were processed by viral 3C protease, as shown by Western blotting, and were antigenic, as revealed by their reactivity in an indirect sandwich-ELISA, and by immunofluorescent assay. The empty capsid-like particles were similar to authentic 75S empty capsids from FMDV in terms of their shape, size and sedimentation velocity, as demonstrated by sucrose gradient centrifugation. Both empty capsid-like particles and some small-sized particles (about 10nm in diameter) were also observed using immunoelectron microscopy. Furthermore, the empty capsid-like particles or intermediates induced high levels of FMDV-specific antibodies in guinea pigs following immunization, and neutralizing antibodies were induced in the second week after vaccination. These recombinant, non-infectious, FMDV empty capsids are potentially useful for the development of new diagnostic techniques and vaccines.
机译:口蹄疫病毒(FMDV)的组装需要蛋白酶3C将P12A多蛋白裂解为单个结构蛋白。在这项研究中,我们构建了重组杆状病毒,该杆状病毒同时从单个启动子表达Asia I FMDV的P12A和3C蛋白的基因。如通过蛋白质印迹所示,在高五昆虫细胞中表达的衣壳蛋白被病毒3C蛋白酶处理,并且通过在间接三明治ELISA中的反应性和免疫荧光分析显示它们是抗原性的。如蔗糖梯度离心所证明的,空衣壳样颗粒的形状,大小和沉降速度与FMDV的真实75S空衣壳相似。使用免疫电子显微镜还观察到了空衣壳状颗粒和一些小颗粒(直径约10nm)。此外,空的衣壳样颗粒或中间体在免疫后在豚鼠中诱导了高水平的FMDV特异性抗体,并且在疫苗接种后的第二周中和了中和抗体。这些重组的,非感染性的,FMDV空衣壳可能对开发新的诊断技术和疫苗有用。

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