Hao Huang, Xiu Li, Zehua Zhang, Min Jia, Hongbo Hu, Zhihong Wu, Zhenhua Zhu, Xiaohong Wan, Hanju Huang
Department of Pathogentic Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan city 430030, PR China.
Genet Vaccines Ther. 2008 Oct 22;6:15. doi: 10.1186/1479-0556-6-15.
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.
在本研究中,我们通过将白细胞介素-2(IL-2)基因与汉坦病毒(HTNV)G2片段DNA进行基因连接,构建了一种新型的嵌合HTNV-IL-2-G2 DNA疫苗质粒,并测试其是否可作为候选疫苗。嵌合基因首先在真核表达系统pcDNA3.1(+)中表达。HTNV-IL-2-G2在COS-7细胞中表达出一种72 kDa的融合蛋白。同时,该融合蛋白保留了其亲本蛋白的活性。此外,用嵌合基因对BALB/c小鼠进行免疫接种。采用酶联免疫吸附测定(ELISA)、体外细胞微量培养中和试验检测免疫接种的BALB/c小鼠的体液免疫反应。采用淋巴细胞增殖试验检测细胞免疫反应。结果表明,嵌合基因可同时诱导产生针对G2糖蛋白和IL-2的特异性抗体。各免疫组小鼠均产生了不同滴度的中和抗体。淋巴细胞增殖试验结果显示,嵌合基因刺激脾细胞对G2和IL-2的刺激指数显著高于其他组。我们的结果表明,基于IL-2的HTNV G2 DNA可诱导针对HTNV G2的体液免疫和细胞免疫反应,可作为HTNV感染的候选DNA疫苗。