肿瘤坏死因子α基因转染人脑胶质瘤浸润淋巴细胞的实验研究  

A Preliminary Study on Transfection of Tumorinfiltrating Lymphocytes from Human Glioma with Tumor Necrosis Factor Alpha Gene

作　　者：陈华群[1] 吴幼章[1] 倪黎[1] 李洪福[1] 王文洪 丁小健[1] 汪承亚[1] 

机构地区：[1]南京医科大学第一附属医院中心实验室

出　　处：《南京医科大学学报（自然科学版）》1998年第3期182-185,共4页Journal of Nanjing Medical University(Natural Sciences)

摘　　要：采用脂质体转染技术将能表达肿瘤坏死因子α（ＴＮＦ－α）基因的重组逆转录病毒质粒ｐＬ（ＴＮＦ－α）ＳＮ导入病毒包装细胞ＰＡ３１７，在细胞培养上清液中得到重组病毒，滴度为１．４×１０９ＣＦＵ／Ｌ。将此病毒上清液感染人脑胶质瘤浸润淋巴细胞（ＴＩＬ），用Ｇ４１８筛选出能表达标记基因ＮｅｏＲ的阳性克隆，ＥＬＩＳＡ法测得ＮｅｏＲ阳性ＴＩＬ培养上清液中存在ＴＮＦ－α，可达５３０ｎｇ／Ｌ，Ｌ９２９依赖细胞法并测出上清液中有ＴＮＦ－α生物学活性。结果表明。To study the availability for transfecting TNF alpha gene into tumor infiltrating lymphocytes (TIL) from human glioma with retroviral vector pLXSN of which was approved by the recombinant DNA advisory committee, NIH, USA. Method: Expressible TNF alpha gene was inserted into pLXSN retroviral vector through BamHⅠ site. The resulting construct was transfected into PA317 packing line with lipofectin method, and high titer (1.4×109 CFU/L) of recombinant virus was obtained. We infected tumorinfiltrating cells isolated from human glioma biopsies with retrovirus and obtained G418 resistant cells. Expressed TNF in culture supernatant of viral infected TIL cells was detectable both by ELISA and L929 dependent cells. The TNFα expressible recombinant virus prepared can infect TIL cells from human glioma driven the TNFα gene expression effectively.

关 键 词：肿瘤坏死因子Α 人脑胶质瘤浸润淋巴细胞 基因转染 

分 类 号：R739.410.5[医药卫生—肿瘤]