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病毒性出血性败血症病毒(VHSV)异源重组抗原蛋白在小鼠体内的免疫反应。

Immune response of heterologous recombinant antigenic protein of viral hemorrhagic septicemia virus (VHSV) in mice.

作者信息

Shin Chunha, Kang Yangjoo, Kim Heui-Soo, Shin Yong Kyoo, Ko Kisung

机构信息

Department of Medicine, College of Medicine, Chung-Ang University, Seoul, South Korea.

Department of Biological Sciences, College of Natural Sciences, Pusan National University, Busan, South Korea.

出版信息

Anim Cells Syst (Seoul). 2019 Feb 8;23(2):97-105. doi: 10.1080/19768354.2019.1575904. eCollection 2019 Apr.

Abstract

Viral hemorrhagic septicemia (VHS) is an important infectious disease in fish worldwide caused by viral hemorrhagic septicemia virus (VHSV). VHSV is the causative agent of serious systemic diseases in fish, affecting a number of teleost fish species. In this study, VHSV glycoprotein (G), including its epitope, as a subunit vaccine candidate, was expressed in tobacco plant (). The recombinant gene, VHSVG, was fused to the immunoglobulin Fc fragment and extended with the endoplasmic reticulum (ER) retention signal (KDEL) to generate VHSVG-FcK. The recombinant expression vector for VHSVG-FcK was transferred into (LBA4404), and plant transformation was conducted . Polymerase chain reaction (PCR) was performed to confirm gene insertion and VHSVG-FcK protein expression was confirmed by immunoblot analysis. VHSVG-FcK protein was successfully purified from tobacco plant leaves. Furthermore, ELISA analysis showed that mice serum immunized with the plant-derived VHSVG-FcK (VHSVGP-FcK) had a high absorbance against VHSVG-FcK, indicating that the plant-derived recombinant subunit vaccine protein VHSVG-FcK can induce immune response. Taken together, this recombinant vaccine protein can be expressed in plant expression systems and can be appropriately assembled to be functional in immunogenicity.

摘要

病毒性出血性败血症(VHS)是一种由病毒性出血性败血症病毒(VHSV)引起的、在全球范围内对鱼类来说都很重要的传染病。VHSV是鱼类严重全身性疾病的病原体,会影响多种硬骨鱼物种。在本研究中,作为亚单位疫苗候选物的VHSV糖蛋白(G),包括其表位,在烟草植株中得以表达。重组基因VHSVG与免疫球蛋白Fc片段融合,并通过内质网(ER)滞留信号(KDEL)进行延伸,以产生VHSVG-FcK。将VHSVG-FcK的重组表达载体转入根癌农杆菌(LBA4404),并进行植物转化。进行聚合酶链反应(PCR)以确认基因插入情况,并通过免疫印迹分析确认VHSVG-FcK蛋白的表达。VHSVG-FcK蛋白成功从烟草叶片中纯化出来。此外,酶联免疫吸附测定(ELISA)分析表明,用植物源VHSVG-FcK(VHSVGP-FcK)免疫的小鼠血清对VHSVG-FcK具有高吸光度,这表明植物源重组亚单位疫苗蛋白VHSVG-FcK能够诱导免疫反应。综上所述,这种重组疫苗蛋白可以在植物表达系统中表达,并且能够适当地组装以在免疫原性方面发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b44/6440531/2b93bede5274/TACS_A_1575904_F0001_OB.jpg

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