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使用重组烟草花叶病毒(TMV)载体在植物中产生的牛疱疹病毒gD蛋白具有真实的抗原性。

Bovine herpes virus gD protein produced in plants using a recombinant tobacco mosaic virus (TMV) vector possesses authentic antigenicity.

作者信息

Pérez Filgueira D M, Zamorano P I, Domínguez M G, Taboga O, Del Médico Zajac M P, Puntel M, Romera S A, Morris T J, Borca M V, Sadir A M

机构信息

School of Biological Sciences, University of Nebraska, Lincoln, NE 68588-0118 , USA.

出版信息

Vaccine. 2003 Oct 1;21(27-30):4201-9. doi: 10.1016/s0264-410x(03)00495-x.

Abstract

A tobacco mosaic virus (TMV)-based vector was utilized for expression of a cytosolic form of the bovine herpesvirus type 1 (BHV-1) protein glycoprotein D (gDc). Nicotiana benthamiana plants were harvested 7 days after inoculation with RNA transcripts derived from the TMV-gDc recombinant virus. Recombinant gDc protein of expected electrophoretic mobility accumulated in inoculated leaves to a concentration of about 20 micrograms/g of fresh leaf tissue. Oil-based vaccines were formulated with crude foliar extracts to immunize mice parentally. After a single injection, animals developed a sustained and specific response to both the isolated gD and native virus particles. Cattle vaccinated with the same gDc containing extracts developed specific humoral and cellular immune responses directed against both the viral gD and BHV-1 particles. Most importantly, animals vaccinated with the plant-produced gDc showed good levels of protection after challenge with the virulent BHV-1. Virus excretion was drastically reduced in these animals, reaching levels comparable to animals vaccinated with a commercial BHV-1 vaccine. The positive immunological characterization obtained for the gDc, indicated that an important part of the natural conformation was retained in the plant recombinant protein.

摘要

一种基于烟草花叶病毒(TMV)的载体被用于表达牛疱疹病毒1型(BHV-1)糖蛋白D(gDc)的胞质形式。在用源自TMV-gDc重组病毒的RNA转录本接种后7天,收获本氏烟草植株。预期电泳迁移率的重组gDc蛋白在接种叶片中积累,浓度约为20微克/克鲜叶组织。用粗叶提取物配制油基疫苗对小鼠进行经皮免疫。单次注射后,动物对分离的gD和天然病毒颗粒产生持续且特异性的反应。用含有相同gDc的提取物接种的牛产生了针对病毒gD和BHV-1颗粒的特异性体液免疫和细胞免疫反应。最重要的是,用植物产生的gDc接种的动物在受到强毒BHV-1攻击后表现出良好的保护水平。这些动物的病毒排泄量大幅减少,达到与接种商业BHV-1疫苗的动物相当的水平。对gDc获得的阳性免疫学特征表明,天然构象的重要部分在植物重组蛋白中得以保留。

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