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莫科拉病毒糖蛋白在杆状病毒中的结构与表达:一种高效重组疫苗

Structure and expression in baculovirus of the Mokola virus glycoprotein: an efficient recombinant vaccine.

作者信息

Tordo N, Bourhy H, Sather S, Ollo R

机构信息

Unité de la Rage, Institut Pasteur, Paris, France.

出版信息

Virology. 1993 May;194(1):59-69. doi: 10.1006/viro.1993.1235.

Abstract

The sequence of the glycoprotein gene of the Mokola virus, the more divergent element of the Lyssavirus genus, has been determined and the predicted protein structure compared to its counterpart in rabies vaccine strains. A global similarity of 54.3% was observed. The divergence affects particularly the rabies antigenic sites involved in the B-cell response. This provides a molecular basis for the absence of cross-protection between Mokola and rabies viruses and argues for the necessity of a specific anti-Mokola vaccine. Toward this goal, a cDNA copy of the glycoprotein gene was cloned into the baculovirus and expressed in Spodoptora frugiperda cells. A recombinant protein was produced in substantial amounts at the surface of the insect cells. Although less strongly glycosylated than the native viral glycoprotein produced in BHK-21 cells, the recombinant protein is antigenically and immunologically similar, it is recognized by specific monoclonal antibodies, and protects mice against an intracerebral challenge with Mokola virus. It therefore constitutes the first experimental genetically engineered vaccine against a rabies-related virus, and fulfills the international standard for protection.

摘要

莫科拉病毒是狂犬病毒属中差异较大的一个成员,其糖蛋白基因序列已被确定,并将预测的蛋白质结构与其在狂犬病疫苗株中的对应结构进行了比较。观察到整体相似度为54.3%。这种差异尤其影响参与B细胞反应的狂犬病抗原位点。这为莫科拉病毒和狂犬病毒之间缺乏交叉保护提供了分子基础,并表明有必要研发一种特定的抗莫科拉疫苗。为实现这一目标,将糖蛋白基因的cDNA拷贝克隆到杆状病毒中,并在草地贪夜蛾细胞中表达。在昆虫细胞表面大量产生了一种重组蛋白。虽然该重组蛋白的糖基化程度不如在BHK - 21细胞中产生的天然病毒糖蛋白,但它在抗原性和免疫性上相似,能被特异性单克隆抗体识别,并能保护小鼠免受莫科拉病毒的脑内攻击。因此,它构成了第一种针对狂犬病相关病毒的实验性基因工程疫苗,并符合国际保护标准。

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