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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 ,脾内注射后能转
Objective To investigate the feasibility and effectiveness of genemodification (GM) fetal spleen transplantation (FLC) in spleen transplantation to mediate cytokine gene therapy and to provide a therapeutic approach for the recovery of patients’ immune response and hematopoietic function after radiotherapy. way. Methods The spleen of FLC mice modified by LacZ or GM-CSF gene were injected into spleen to observe their biological behavior. Results ① The infection rate (MOI) was 50 and the transfection efficiency was 80% ~ 85% when the infection time was 2h. ② The FACS analysis showed that the gene transfer system could inhibit the cell cycle, apoptosis Cell ratio and CD34 + cell content. (3) The number of spleen colony forming unit (CFU S8) on 8d after injection of FLC GM in mice was significantly increased, suggesting that GM CSF gene transfection can promote the expression of FLC in FLC The proliferation, differentiation and in vivo implantation of late-oriented stem cells were observed in the late stage. (2) 20% ~ 25% of cells transplanted into the liver 2 hours after transplanted with 111In-labeled FLC, %, And continue until the detection of the first 5 days, only about 2 2% of the liver cells remain in the spleen; ⑤ serum secretion level in spleen 48h after transplantation up to (35 6 ± 5 8) pg / ml, (P <0.05). (6) Allogeneic FLC GM spleen transplantation induced host delayed type hypersensitivity (DTH) intensity was significantly weaker than allogeneic spleen cells subcutaneous sensitized group (control group). Conclusion Gene-modified FLC can maintain its original biological properties, after injection of spleen can turn