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杆状病毒表达的兔出血症病毒(捷克斯洛伐克毒株V-351)衣壳蛋白的自组装、抗原性及免疫原性

Self-assembly, antigenicity, and immunogenicity of the rabbit haemorrhagic disease virus (Czechoslovakian strain V-351) capsid protein expressed in baculovirus.

作者信息

Nagesha H S, Wang L F, Hyatt A D, Morrissy C J, Lenghaus C, Westbury H A

机构信息

CSIRO Australian Animal Health Laboratory, Geelong, Victoria.

出版信息

Arch Virol. 1995;140(6):1095-1108. doi: 10.1007/BF01315418.

Abstract

Rabbit haemorrhagic disease virus (RHDV) capsid protein was expressed in a baculovirus system. Analysis of the expressed product showed that the recombinant protein, which is 60 kDa in size, was antigenic as revealed by its reactions in ELISA and Western blot with the antibodies raised against RHDV. Direct electron microscopy of the cell culture supernatant and the purified protein demonstrated that the capsid protein expressed in insect cells self-assembled to form empty virus-like particles (VLP) which are similar in size and morphology to that of native virus. These particles were immunoreactive with polyclonal anti-RHDV antibodies and with four monoclonal antibodies which recognise conformational epitopes of the virus. The results indicated that the VLPs were morphologically and antigenically indistinguishable from native virus. The recombinant VLPs induced high levels of RHDV-specific antibodies in rabbits and mice following immunisation. The immune response to the VLPs protected the rabbits following challenge with the virulent RHDV. In haemagglutination assays, the VLPs bound to human red blood cells similar to the native virus particles. The recombinant protein and or VLPs is suitable for the development of a rapid, sensitive and reliable test for detection of antibodies to RHDV and for use as a vaccine for domestic rabbits.

摘要

兔出血症病毒(RHDV)衣壳蛋白在杆状病毒系统中表达。对表达产物的分析表明,该重组蛋白大小为60 kDa,在ELISA和Western blot中与针对RHDV产生的抗体反应,显示具有抗原性。对细胞培养上清液和纯化蛋白进行直接电子显微镜观察表明,昆虫细胞中表达的衣壳蛋白自组装形成空的病毒样颗粒(VLP),其大小和形态与天然病毒相似。这些颗粒与多克隆抗RHDV抗体以及四种识别病毒构象表位的单克隆抗体发生免疫反应。结果表明,VLP在形态和抗原性上与天然病毒无法区分。重组VLP在免疫后在兔和小鼠中诱导产生高水平的RHDV特异性抗体。对VLP的免疫反应在兔受到强毒RHDV攻击后起到了保护作用。在血凝试验中,VLP与天然病毒颗粒一样与人红细胞结合。重组蛋白和/或VLP适用于开发一种快速、灵敏且可靠的检测RHDV抗体的试验,并用作家兔疫苗。

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