摘要
去除供者骨髓中的T淋巴细胞可有效防止移植物抗宿主病 (GVHD)的发生 .用含大肠杆菌胞嘧啶脱氨酶 (EC CD)基因的高滴度逆转录病毒 (1 5× 10 5CFU/ml)感染小鼠T淋巴细胞 ,G418筛选 ,获得稳定表达CD基因的T/pCD2 细胞 .经PCR和RT PCR方法检测证明CD基因已成功地导入T淋巴细胞中并有效表达 .分别用不同浓度的 5 FCyt作用于T/pCD2 及T淋巴细胞 ,光镜下观察不同时间细胞数目变化及MTT法检测细胞活性 .结果表明 ,5 FCyt浓度大于 1μmol/L时 ,即对T/pCD2 细胞有显著的杀伤作用 ,而对正常T淋巴细胞基本无毒性 ,且T/pCD2 细胞在加入药物后生存时间 (3～ 5d)明显短于未转染的T淋巴细胞 (大于 14d)
T lymphocyte depletion of the marrow graft can prevent graft versus host disease(GVHD) effectively. T lymphocytes of mice were transfected with high titer (1 5×10 5 CFU/ml) retrovirus supernatant, and by selection with G418 positive cell clones (T/pCD 2) were obtained. PCR and RT PCR showed that CD gene was transferred into the T lymphocyte and expressed successfully. T lymphocyte and T/pCD 2 cells were exposed to different doses of 5 FCyt for various hours,and were then observed under light microscope and the viability of these cells was determined by MTT colorimetric cell proliferation assay. The results showed that retrovirus mediated CD gene transferred to T/ pCD 2 cells could confer them high sensitivity to 5 FCyt, and T/ pCD 2 cells exposed to 5 FCyt (>1 μmol/L) can be killed. But 5 FCyt was not harmful to normal T lymphocyte cells. The survival time of T/ pCD 2 cells in the presence of 5 FCyt(3～5 d) was significantly shorter than that of T lymphocyte( >14 d ).
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2000年第5期524-528,共5页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金!(396 0 0 0 6 6 ) .
