CEA迷你基因串联体基因疫苗的构建及其免疫学活性研究

发布时间：2018-05-31 22:36

本文选题：癌胚抗原 + 迷你基因　；参考：《吉林大学》2008年博士论文

【摘要】： CEA作为肿瘤相关抗原,因其在正常组织中极有限的表达,以及广泛出现在上皮源性肿瘤细胞的过量表达而成为肿瘤生物治疗研究的热点。本研究选择了包含了两个CEA HTL表位(CEA625-639和CEA653-667)的迷你基因CEA625-667来制作串联体基因疫苗。首先利用CEA625-667短肽诱导的脾细胞增殖实验确认了CEA625-667在小鼠体内的抗原性之后,进一步应用同尾酶连接方法成功地构建了含有CEA625-667迷你基因三串联体的DNA疫苗pcDNA-triCEA625-667。并通过动物实验对其免疫学活性进行研究。动物免疫研究的特异性淋巴细胞增殖实验及抗原特异性抗体的检测结果表明迷你基因疫苗可诱导辅助性T细胞增殖。Th1/ Th2类细胞因子含量测定结果分析提示了本研究所选用的高亲和力Th表位使被免疫机体T细胞趋向于Th1效应。且迷你基因三倍体疫苗所引发的增殖效应以及释放细胞因子的水平均高于迷你基因一倍体,说明我们所采用的将目的基因多倍串联的抗原改造方式起到了增强免疫效应的作用。本研究探讨了迷你基因疫苗设计的新方法,对表位串联疫苗的构建及以HTL表位为基础的疫苗的功能进行了深入研究,为有关CEA疫苗的研制与临床前实验研究奠定了基础。
[Abstract]:As a tumor associated antigen, CEA has become a hot topic in tumor biotherapy because of its limited expression in normal tissues and its overexpression in epithelial-derived tumor cells. In this study, a mini gene CEA625-667 containing two CEA HTL epitopes, CEA625-639 and CEA653-667, was selected to prepare the tandem gene vaccine. At first, the antigenicity of CEA625-667 in mice was confirmed by CEA625-667 short peptide induced splenocyte proliferation test, and then the DNA vaccine pcDNA-triCEA625-667 containing CEA625-667 mini-gene tritandem was successfully constructed by isotail enzyme linking method. Its immunological activity was studied through animal experiments. The results of specific lymphocyte proliferation test and antigen-specific antibody detection in animal immunological study showed that mini gene vaccine could induce T cell proliferation. Th1 / Th2 cytokines content analysis suggested that this study could be carried out. The selected high affinity Th epitopes make the immune T cells tend to the Th1 effect. The proliferation effect and the release level of cytokines induced by the mini gene triploid vaccine were higher than that of the mini gene haploidy, which indicated that the antigenic modification method of multifold tandem of the target gene played a role in enhancing the immune response. In this study, a new method for the design of mini gene vaccine was discussed, and the construction of epitope tandem vaccine and the function of HTL epitope based vaccine were studied in depth, which laid a foundation for the development of CEA vaccine and preclinical experimental research.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R392.1

【参考文献】