Rümenapf T, Stark R, Meyers G, Thiel H J
Federal Research Centre for Virus Diseases of Animals, Tübingen, Federal Republic of Germany.
J Virol. 1991 Feb;65(2):589-97. doi: 10.1128/JVI.65.2.589-597.1991.
A cDNA fragment covering the genomic region that encodes the structural proteins of hog cholera virus (HCV) was inserted into the tk gene of vaccinia virus. Expression studies with vaccinia virus/HCV recombinants led to identification of HCV-specific proteins. The putative HCV core protein p23 was demonstrated for the first time by using an antiserum against a bacterial fusion protein. The glycoproteins expressed by vaccinia virus/HCV recombinant migrated on sodium dodecyl sulfate-gels identically to glycoproteins precipitated from HCV-infected cells. A disulfide-linked heterodimer between gp55 and gp33 previously detected in HCV-infected cells was also demonstrated after infection with the recombinant virus. The vaccinia virus system allowed us to identify, in addition to the heterodimer, a disulfide-linked homodimer of HCV gp55. The vaccinia virus/HCV recombinant that expressed all four structural proteins induced virus-neutralizing antibodies in mice and swine. After immunization of pigs with this recombinant virus, full protection against a lethal challenge with HCV was achieved. A construct that lacked most of the HCV gp55 gene failed to induce neutralizing antibodies but induced protective immunity.
将覆盖编码猪瘟病毒（HCV）结构蛋白的基因组区域的cDNA片段插入痘苗病毒的tk基因中。对痘苗病毒/HCV重组体进行的表达研究导致鉴定出HCV特异性蛋白。首次使用针对细菌融合蛋白的抗血清证实了推定的HCV核心蛋白p23。痘苗病毒/HCV重组体表达的糖蛋白在十二烷基硫酸钠凝胶上的迁移情况与从HCV感染细胞中沉淀的糖蛋白相同。在用重组病毒感染后，也证实了先前在HCV感染细胞中检测到的gp55和gp33之间的二硫键连接的异二聚体。痘苗病毒系统使我们能够除了鉴定异二聚体外，还鉴定出HCV gp55的二硫键连接的同二聚体。表达所有四种结构蛋白的痘苗病毒/HCV重组体在小鼠和猪中诱导出病毒中和抗体。用这种重组病毒免疫猪后，实现了对HCV致死性攻击的完全保护。一个缺少大部分HCV gp55基因的构建体未能诱导中和抗体，但诱导了保护性免疫。
