Genetic immunization with Hantavirus vaccine combining expression of G2 glycoprotein and fused interleukin-2.

Huang Hao, Li Xiu, Zhang Zehua, Jia Min, Hu Hongbo, Wu Zhihong, Zhu Zhenhua, Wan Xiaohong, Huang Hanju
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引用次数: 2

Abstract

In this research, we developed a novel chimeric HTNV-IL-2-G2 DNA vaccine plasmid by genetically linking IL-2 gene to the G2 segment DNA and tested whether it could be a candidate vaccine. Chimeric gene was first expressed in eukaryotic expression system pcDNA3.1 (+). The HTNV-IL-2-G2 expressed a 72 kDa fusion protein in COS-7 cells. Meanwhile, the fusion protein kept the activity of its parental proteins. Furthermore, BALB/c mice were vaccinated by the chimeric gene. ELISA, cell microculture neutralization test in vitro were used to detect the humoral immune response in immunized BALB/c mice. Lymphocyte proliferation assay was used to detect the cellular immune response.- The results showed that the chimeric gene could simultaneously evoke specific antibody against G2 glycoprotein and IL-2. And the immunized mice of every group elicited neutralizing antibodies with different titers. Lymphocyte proliferation assay results showed that the stimulation indexes of splenocytes of chimeric gene to G2 and IL-2 were significantly higher than that of other groups. Our results suggest that IL-2-based HTNV G2 DNA can induce both humoral and cellular immune response specific for HTNV G2 and can be a candidate DNA vaccine for HTNV infection.

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汉坦病毒疫苗联合表达G2糖蛋白和融合白细胞介素-2的遗传免疫。
本研究通过IL-2基因与G2片段DNA的遗传连接,构建了一种新型嵌合HTNV-IL-2-G2 DNA疫苗质粒,并对其是否可作为候选疫苗进行了试验。嵌合基因首次在真核表达系统pcDNA3.1(+)中表达。HTNV-IL-2-G2在COS-7细胞中表达72kda的融合蛋白。同时,融合蛋白保持了亲本蛋白的活性。此外,用嵌合基因接种BALB/c小鼠。采用ELISA法、体外细胞微培养中和试验检测免疫后BALB/c小鼠体液免疫应答。淋巴细胞增殖法检测细胞免疫应答。-结果表明,嵌合基因可同时产生针对G2糖蛋白和IL-2的特异性抗体。各组免疫小鼠均产生不同滴度的中和抗体。淋巴细胞增殖试验结果显示,嵌合基因的脾细胞对G2和IL-2的刺激指数显著高于其他各组。我们的研究结果表明,基于il -2的HTNV G2 DNA可以诱导HTNV G2特异性的体液和细胞免疫反应,可以作为HTNV感染的候选DNA疫苗。
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