TNF基因转导肿瘤浸润淋巴细胞的方法学研究  

Preliminary Study on Tnmor-infiltrating Lymphocytes Transfected with Tumor Necrosis Factor

作　　者：李彪如[1] 徐铿[1] 钱关祥[1] 张希衡[1] 童善庆[1] 陈诗书[1] 

机构地区：[1]上海第二医科大学基因治疗研究中心

出　　处：《上海第二医科大学学报》1995年第3期185-189,共5页Acta Universitatis Medicinalis Secondae Shanghai

摘　　要：在总结Ｒｏｓｅｎｂｅｒｇ建立的ＴＮＦ－ＮｅｏＲ缺陷型逆转录病毒转染肿瘤浸润细胞（ＴＩＬ）的基础上，选用美国国家卫生院重组ＤＮＡ咨询委员会（ＲＡＣ）批准的逆转录病毒（Ｍｏ－ＭｕＬＶ）的衍生物作载体，对肿瘤坏死因子转导ＴＩＬ进行研究。实验结果提示：ＴＩＬ处于静息期与对数生长后期，当ＴＮＦ－ＮｅｏＲ缺陷型逆转录病毒转导ＴＩＬ细胞时，ＴＩＬ细胞增殖受到影响；最佳转导时间为ＴＩＬ分离培养后９～２５ｄ。各病毒感染复数值病毒转导对ＴＩＬ增殖倍数与生存天数影响不大，但病毒感染复数值越大，转导后ＴＩＬ起始呈抑制现象越大；然而Ｇ_（４１８）筛选后证实，病毒感染复数值越大，转导成功率越大。Ｌ_（９２９）细胞检测转导后ＴＩＬ呈高表达，ＰＣＲ检测ＴＮＦ目的基因已插入宿主ＤＮＡ中。In the paper,we selected Mo-MuLV derivative approved by the Recombinant DNA Advisory Committee (RAC)of the NIH,USA, as vector and tumor-infiltrating lymphocytes as vehicles to study relationships between the TNF gene and TILs. The results showed that TILs proliferated slowly when TILs were transfected in the resting phase or latter log-growth phase .The better time to transfect the yector is when TILs were isolated and cultured from 9 to 25 days. The study on relationship between multiplicity of infection (MOI)and TILs being transfected showed that the total TIL expansion-fold and survival time of different MOI values confused with the control group were not significantly different (P>0.05)but the more MOI is,the more inhibitive is in the first period after transfection, The more MOI is, the greater TILs having been transfected are under G_(418) selection. The polymerase chain reaction and L(929) cell test all showed the TNF gene integration and expression.

关 键 词：肿瘤浸润 淋巴细胞 转导 肿瘤坏死因子 方法学 

分 类 号：R730.3[医药卫生—肿瘤] R394.8[医药卫生—临床医学]