摘要
用梯形DNA电泳与Gimsa染色研究人T细胞瘤细胞株Jurkat在转导编码人 6A8α 甘露糖苷酶基因的反向cDNA后 ,对抗Fas抗体诱导凋亡敏感性的变化 ,并用间接免疫荧光染色 (流式细胞仪 )检测细胞表面Fas分子的表达。结果表明 ,抗Fas抗体诱导细胞 2 4h后 ,野生型及转导空载载体的Jurkat细胞出现明显的梯形DNA ,两者之间无明显差异 ,但转导反向 6A8cDNA的细胞未见明显的梯形DNA。Gimsa染色结果显示转导反向 6A8cDNA的Ju rkat细胞中凋亡细胞数明显减少 ,而转导空载载体则无影响。Jurkat细胞为Fas(CD95 ,Apo 1)全阳性细胞 ,转导反向 6A8cDNA或空载载体对Fas表达阳性率与Fas表达强度无明显影响。以上结果提示 ,转导编码 6A8α 甘露糖苷酶基因的反向cDNA使人T细胞株Jurkat对抗人Fas抗体诱导的凋亡作用产生耐受。
DNA ladder and Giemsa's staining were used to detect changes of susceptibility of the Jurkat cells transducted with rAAV reverse 6A8 α mannosidase cDNA to apoptosis induction with mAb anti Fas Indirect immunofluorescent staining was performed to detect the Fas expression on cell surface DNA ladder was observed in wild type and mock transducted Jurkat cells, but not in the rAAV reverse 6A8 cDNA transducted ones after 24 h treatment with mAb anti Fas Giemsa's staining showed a number of dying and died cells (with apoptosis bodies) in wild type and the mock transducted ones upon anti Fas antibody treatment Mock transduction did not affect the apoptosis induction However, in the rAAV reverse 6A8 cDNA transducted cells, few apoptotic cells could be found 100% cells in each group expressed Fas molecular The data indicated that Jurkat cells became resistant to apoptosis induction by anti Fas antibody upon transduction with a reverse cDNA encoding human 6A8 α mannosidase and the transduction did not affect the expression of Fas molecular
出处
《基础医学与临床》
CSCD
北大核心
2001年第2期136-140,共5页
Basic and Clinical Medicine
基金
国家自然科学基金!重点项目 (396 30 30 0 )
