乙肝病毒DNA疫苗双表达载体的构建及其诱导小鼠的免疫应答

发布时间：2018-06-20 01:42

  本文选题：HBV + DNA　； 参考：《吉林大学》2006年硕士论文

【摘要】：乙型肝炎病毒(HBV)感染是严重危害人类健康的传染病之一,基因重组乙肝疫苗是目前为止预防乙肝病毒感染的有力手段。然而在接种乙肝疫苗的健康人群中约有14%的人表现为对疫苗抗原的无应答或弱应答,不能产生保护力水平的抗体,因此,研制新型乙肝疫苗及改进现有乙肝疫苗的免疫活性成为目前急需解决的问题。有关研究结果表明,乙肝病毒DNA疫苗可在免疫动物体内诱导出持续的体液免疫和细胞免疫反应,并能打破机体对HBV的免疫耐受,可兼作预防性和治疗性疫苗,是未来新型疫苗的发展方向之一。 本研究将微小病毒内部核糖体进入位点(IRES)基因克隆到核酸疫苗载体pVAXⅠ多克隆位点处,再将乙肝病毒表面抗原(HBsAg)基因和粒细胞-巨噬细胞集落刺激因子(GM-CSF)基因依次克隆到IRES的上、下游多克隆位点处,构建出核酸疫苗双表达载体pHIG。 用双表达质粒pHIG免疫BaLB/C小鼠,同时设置阴性对照组注射pVAXⅠ空质粒,并分别于第2、4周后加强免疫各1次。 结果显示,试验组在免疫1周时开始有HBsAb的产生,阴性对照组未测到HBsAb。含有乙肝病毒HbsAg基因与GM-CSF基因的双表达质粒能引起小鼠特异性体液免疫应答并可使免疫效果提高1倍。
[Abstract]:Hepatitis B virus (HBV) infection is one of the most serious infectious diseases, and recombinant hepatitis B vaccine is a powerful method to prevent hepatitis B virus infection. However, about 14% of the healthy people who were vaccinated with hepatitis B vaccine showed no or weak response to the vaccine antigen and could not produce antibodies at the protective level. It is urgent to develop new hepatitis B vaccine and improve the immune activity of existing hepatitis B vaccine. The results show that HBV DNA vaccine can induce sustained humoral and cellular immune responses in immunized animals, break the immune tolerance to HBV, and can be used as both prophylactic and therapeutic vaccines. It is one of the development directions of the new vaccine in the future. In this study, the internal ribosome entry site (IRES) gene of parvovirus was cloned to the polyclonal site of nucleic acid vaccine vector pVAX 鈪，

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