Preparation and immunogenic properties of a recombinant West Nile subunit vaccine
Preparation and immunogenic properties of a recombinant West Nile subunit vaccine
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DOI：
10.1016/j.vaccine.2006.08.018
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发表时间：
2007-01-05
期刊：
影响因子：
5.5
通讯作者：
Weeks-Levy, Carolyn
中科院分区：
文献类型：
--
作者：
Lieberman, Michael M.;Clements, David E.;Weeks-Levy, Carolyn
虽然几种西尼罗河疫苗正在开发中，但还没有一种疫苗可用于人类。本研究在果蝇S2细胞表达系统中表达西尼罗河病毒(West Nile Virus，WNV)囊膜蛋白(E)和非结构蛋白1(NS1)，旨在研制人用疫苗。含有膜锚定部分的E蛋白的20%的C末端被删除，从而允许截断的蛋白(80E)有效地分泌到细胞培养基中。用黄病毒囊膜蛋白组特异性(80E)或黄病毒NS1组特异性(NS1)单抗进行免疫亲和层析(IAC)纯化。纯化的蛋白质产量高，并与佐剂配方一起用于接种小鼠。分别用空斑减少中和试验和酶联免疫吸附试验、淋巴细胞增殖试验和细胞因子产生试验检测小鼠的体液免疫应答和细胞免疫应答。结果表明，80E蛋白和NS1蛋白均能诱导小鼠产生高滴度的EL ISA抗体和中和抗体。免疫小鼠脾细胞在疫苗抗原刺激下体外培养，表现出良好的增殖能力和细胞因子(干扰素-γ、IL-4、IL-5、IL-10)的产生。抗原刺激的淋巴细胞增殖和细胞因子的产生水平与有丝分裂原(植物血凝素或商陆)刺激的水平相当，表明细胞反应也很强劲。这些发现令人鼓舞，有必要进行进一步的体内研究，以确定候选西尼罗河病毒疫苗的保护效果。(C)2006爱思唯尔有限公司。保留所有权利。
While several West Nile vaccines are being developed, none are yet available for humans. In this study aimed at developing a vaccine for humans, West Nile virus (WNV) envelope protein (E) and non-structural protein 1 (NS1) were produced in the Drosophila S2 cell expression system. The C-terminal 20% of the E protein, which contains the membrane anchor portion, was deleted, thus allowing for efficient secretion of the truncated protein (80E) into the cell culture medium. The proteins were purified by immunoaffinity chromatography (IAC) using monoclonal antibodies that were flavivirus envelope protein group specific (for the 80E) or flavivirus NS1 group specific (for NS1). The purified proteins were produced in high yield and used in conjunction with adjuvant formulations to vaccinate mice. The mice were tested for both humoral and cellular immune responses by a plaque reduction neutralization test and ELISA, and by lymphocyte proliferation and cytokine production assays, respectively. The results revealed that the 80E and the NS1 proteins induced both high-titered ELISA and neutralizing antibodies in mice. Splenocytes from immunized mice, cultured in vitro with the vaccine antigens as stimulants, showed excellent proliferation and production of cytokines (IFN-gamma, IL-4, IL-5, and IL-10). The level of antigen-stimulated lymphocyte proliferation and cytokine production was comparable to the level obtained from mitogen (phytohemagglutinin or pokeweed) stimulation, indicating a robust cellular response as well. These findings are encouraging and warrant further in vivo studies to determine the protective efficacy of the WNV vaccine candidate. (c) 2006 Elsevier Ltd. All rights reserved.