脾内移植基因修饰鼠胎肝细胞的生物学特征  

The biocharacterization of intrasplenic transplantation of gene modified mouse fetal liver cells

作　　者：李士玉[1] 弭静[2] 曹雪涛[2] 杨忠广[3] 朱学军[2] 张明徽[2] 

机构地区：[1]山东省千佛山医院消化科,250014 [2]第二军医大学免疫学教研室 [3]山东医科大学附属医院泌尿科

出　　处：《中华消化杂志》2001年第7期400-403,共4页Chinese Journal of Digestion

摘　　要：目的　探讨基因修饰 (genemodification ,GM )胎肝细胞 (fetallivercells ,FLC)脾内移植途径介导细胞因子基因治疗的可行性和有效性 ,为肿瘤放疗病人免疫和造血功能的恢复提供一种治疗手段和途径。方法　将经LacZ或粒 /巨噬细胞集落刺激因子 (GM CSF)基因体外修饰的小鼠FLC脾内注射后 ,观察其生物学行为。结果　①病毒 /细胞感染比率 (MOI)为 5 0 ,转染时间为 2h时 ,能获得 80 %～85 %的转染效率 ;②通过FACS分析 ,基因转移体系对FLC群体的细胞周期、凋亡细胞比例以及CD34 +细胞含量无明显影响 ;③照射小鼠尾静脉注射FLC GM后 ,其 8d脾克隆形成单位 (CFU S8)数目明显升高 ,提示GM CSF基因转染有助于促进FLC中较晚期定向干细胞的增殖、分化以及体内植入 ;④ 111In标记的FLC脾内移植后 2h即有 2 0 %～ 2 5 %细胞转位于肝脏 ,48h肝内移入量最高 ,达 5 0 %～ 5 5 % ,并持续至检测第 5天 ,只有 2 2 %左右的肝细胞滞留于脾脏 ;⑤血清分泌水平在脾内移植后 48h最高 ,达(35 6± 5 8)pg/ml,此时行脾切除 ,GM CSF仍具有一定量的表达水平 ;⑥同种异体FLC GM脾内移植引起的宿主迟发型变态反应 (DTH)强度明显弱于同种异体脾细胞皮下致敏组 (对照组 )。结论　基因修饰的FLC能够保持其原有的生物学性质 。Objective To develop a genetically modified fetal liver cells (FLC) based transplantation system that can release therapeutic levels of hematopoietic growth factors into the system circulation which can facilitate treatment of patient receiving cytokine therapy following chemotherapy. Method Examine adeno virus mediated gene transfer to isolated murine FLC and evaluate the biocharacterization of intrasplenic transplantation of gene modified murine FLC. Results Substantial transfection rate of 80%～85% were achieved at a ratio of 50 for 2 hr of exposure. Gene modified FLC (FLC GM) labeled with 111 In were injected into the allogenic mice, spleen, the %ID/g of liver was 20%～25% at 24 hr and 50%～55% at 48 hr after transplantation. In addition, serum concentration of GM CSF in mice with intrasplenic transplantation reached its maximum at 48 hr [(356 ±58 ) pg/ml]. Conclusion Intrasplenic transplantation of FLC GM can be predominantly localized in liver and spleen, and engraft rapidly and maintain normal function, which represent a critical step toward successfully accomplishing liver directed gene therapy.

关 键 词：脾内移植 基因修饰 胎肝细胞 生物学行为 肿瘤 动物实验 治疗 

分 类 号：R575[医药卫生—消化系统]