新型自杀基因系统修饰供者T细胞防治移植物抗宿主病的基础研究

发布时间：2018-04-11 11:00

本文选题：酵母胞嘧啶脱氨酶 + 移植物抗宿主病　；参考：《吉林大学》2005年硕士论文

【摘要】：目的:探讨转染新型自杀基因YCD 的T 淋巴细胞其体外功能及对前体药物的敏感性,为其防治GVHD 提供理论依据。方法:①脂质体法转染瞬时包装细胞293T 制备含自杀基因YCD 的逆转录病毒;②将逆转录病毒导入T 细胞,并进行筛选、扩增,获得转基因T 细胞;③应用PCR 方法检测转基因T 细胞YCD 基因的表达,ELISA 方法检测细胞因子分泌情况,MTT 法检测其杀伤功能及对前体药物的敏感性。结果:①成功将自杀基因YCD 转入293T 细胞,基因染率可达38.2%;②PCR 方法可检测到转基因T 细胞中YCD 基因的表达; ③筛选出的转基因T 细胞其增殖功能正常,与正常T 细胞相比,对K562 细胞的细胞毒作用无改变,细胞因子分泌功能正常;④体外实验证实转基因T 细胞可被38μmol/L 的前体药物5-FC 全部杀伤。结论:①本实验利用pMSCV-YCD-Puro 载体,成功的将YCD 基因导入人T细胞;②转基因T 细胞的增殖、细胞因子分泌功能及细胞毒作用未受影响;③转基因T 细胞对前体药物敏感,可被高度杀伤。
[Abstract]:Aim: to investigate the function of T lymphocytes transfected with novel suicide gene YCD in vitro and its sensitivity to precursor drugs in order to provide theoretical basis for the prevention and treatment of GVHD.Methods Twain 1 liposome method was used to transfect 293T of transient packaging cells to produce retrovirus 2 containing suicide gene YCD into T cells. The T cells were screened and amplified to obtain transgenic T cells.3The expression of YCD gene in transgenic T cells was detected by PCR method. The cytokine secretion was detected by Elisa. The cytotoxicity and sensitivity to precursor drugs were detected by MTT assay.Results the suicide gene YCD was successfully transferred into 293T cells. The expression of YCD gene in transgenic T cells could be detected by 2PCR, and the proliferative function of the screened transgenic T cells was normal, compared with that of normal T cells.The cytotoxicity of K562 cells was not changed and the cytokine secretion function was normal in vitro. The results showed that the transgenic T cells could be completely killed by 38 渭 mol/L precursor drug 5-FC.Conclusion in this experiment, pMSCV-YCD-Puro vector was used to successfully transfer YCD gene into human T cell line 2 transgenic T cell proliferation, cytokine secretion function and cytotoxicity were not affected.Can be highly killed.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392

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