以HBcAg-preS1表位融合蛋白为基础的新型HBV治疗性疫苗的构建、表达和免疫活性研究

发布时间：2018-05-21 02:08

  本文选题：HBV + HBcAg　； 参考：《吉林大学》2006年博士论文

【摘要】：乙肝是一种严重威胁人类健康的传染病,目前临床治疗仍没有特效药物。许多研究证实,细胞免疫和体液免疫在阻止HBV感染和病毒清除方面起到了关键的作用。因此,开发能特异增强机体细胞免疫和/或体液免疫的乙肝治疗性药物是该领域研究的热点。 乙肝核心抗原(HBcAg)由于其独特的物理结构和免疫学性质而被人们认为是一种良好的分子载体。本研究利用基因工程技术对HBcAg基因e1区进行了修饰,使其变为能够良好递呈外源基因的分子载体。将前S1表位基因插入该区,构建了含有HBcAg和前S1表位融合基因的原核表达载体pBTcs1。在大肠杆菌中表达后,融合蛋白BTcs1经蔗糖密度梯度超速离心纯化。蛋白鉴定结果显示,融合蛋白BTcs1分子量约为28 kDa,能够与抗-preS1抗体和抗-HBc抗体特异杂交,且能够自主组装成病毒样颗粒(VLPs)结构,颗粒直径约为30-34 nm。融合蛋白BTcs1免疫Balb/c小鼠后,能够诱导出高滴度的保护抗体和Th1型免疫反应,表明BTcs1可能发展成为一种兼有预防和治疗作用的新型HBV疫苗。
[Abstract]:Hepatitis B is a serious threat to human health infectious disease, the current clinical treatment is still no special drugs. Many studies have confirmed that cellular and humoral immunity play a key role in preventing HBV infection and virus clearance. Therefore, the development of hepatitis B therapeutic drugs that can specifically enhance cellular and / or humoral immunity is a hot topic in this field. Hepatitis B core antigen (HBcAg) is considered to be a good molecular carrier because of its unique physical structure and immunological properties. In this study, the gene engineering technique was used to modify the e1 region of HBcAg gene so that it could be transformed into a molecular vector capable of presenting foreign genes well. The prokaryotic expression vector pBTcs1 containing HBcAg and preS1 epitope fusion gene was constructed by inserting preS1 epitope gene into the region. After expression in E. coli, the fusion protein BTcs1 was purified by sucrose density gradient ultracentrifugation. The results of protein identification showed that the molecular weight of the fusion protein BTcs1 was about 28kDa. the fusion protein could be hybridized with anti-preS1 antibody and anti-HBc antibody. The fusion protein could be assembled into virus-like particles with a diameter of about 30-34 nm. The fusion protein BTcs1 immunized Balb/c mice could induce high titers of protective antibody and Th1 type immune response, indicating that BTcs1 might develop into a novel HBV vaccine with both preventive and therapeutic effects.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R392

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本文编号：1917253