含SFV RNA复制子的核酸疫苗的构建及实验免疫研究

发布时间：2018-02-14 01:59

本文关键词： SFV RNA复制子核酸疫苗构建抑瘤作用　出处：《长春理工大学》2008年硕士论文　论文类型：学位论文

【摘要】： 本研究构建了两种含塞姆利基森林病毒(semliki forest virus,SFV) RNA复制子的核酸疫苗pIRSFV-HN和pIRSFV-Apoptin,限制性酶切鉴定正确。采用RT-PCR、Western blotting和IFA检测了外源基因在HepG-2肿瘤细胞内的表达,结果显示外源基因可以在HepG-2肿瘤细胞中有效表达。采用AO/EB染色、DAPI染色和MTT染色方法研究其体外抑瘤作用,结果表明pIRSFV-HN和pIRSFV-Apoptin能够有效杀伤HepG-2肿瘤细胞,最大杀伤率分别为55.43%和53.55%。本研究复制了C57BL/6小鼠荷H22肿瘤模型,瘤内注射pIRSFV-HN和pIRSFV-Apoptin,通过检测特异性CTL活性和T细胞亚类研究其体内抑瘤作用,结果表明pIRSFV-HN和pIRSFV-Apoptin可以诱导较强的CTL反应,并促使细胞免疫趋向Th1优势。
[Abstract]:This research constructs containing two kinds of Semliki Forest virus (Semliki forest, virus, SFV) RNA replicon based nucleic acid vaccine pIRSFV-HN and pIRSFV-Apoptin restriction enzyme digestion. The RT-PCR, Western blotting and IFA to detect the expression of exogenous gene in HepG-2 tumor cells, results showed that the exogenous gene can be effectively expressed in HepG-2 tumor cells. Using AO/EB staining, DAPI staining and MTT staining method to study its antitumor effect in vitro, the results showed that pIRSFV-HN and pIRSFV-Apoptin can effectively kill HepG-2 tumor cells, the maximum killing rate was 55.43% and 53.55%. respectively this study replicated the C57BL/6 mice bearing H22 tumor model, intratumoral injection of pIRSFV-HN and pIRSFV-Apoptin, through the detection of specific CTL activity and T cell subsets of the in vivo antitumor effect, the results showed that pIRSFV-HN and pIRSFV-Apoptin can induce strong CTL responses, and promote Cell immunization tends to Th1 advantage.

【学位授予单位】：长春理工大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

【参考文献】