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为探讨基因修饰的肝细胞经脾内移植途径介导肝脏基因治疗的可行性和有效性,体外用NeoR基因或IL-2基因修饰小鼠正常胚胎肝细胞BNL CL.2,经脾内移植至正常同系小鼠体内(2×10~6/只),观察NeoR和IL-2基因在不同脏器的表达.结果发现脾内移植NeoR基因修饰的肝细胞后24h,即可通过RT-PCR在肝脏中检测出NeoR基因mRNA的表达,持续表达11周以上;此外,NeoR基因在脾脏中短暂表达(24h至1周),在肺组织中也有一过性表达(48～96h).脾内移植IL-2基因修饰的肝细胞后,肝脏中可检测到稳定表达的IL-2mRNA(24h至11周),外周血中维持一定水平(5～40pg/mL)的IL-2,能增强肝脏Kupffer细胞Ia抗原的表达及脾细胞的NK杀伤活性.提示基因修饰的肝细胞脾内移植后能定向分布至肝脏,并同化入肝组织中长期存活,有效地表达外源基因,可成为肝脏靶向性基因治疗的可行途径.
To investigate the feasibility and effectiveness of genetically modified hepatocytes mediated by spleen transplantation in liver gene therapy, NeoR gene or IL-2 gene was used to modify the normal mouse embryonic liver cells BNL CL.2, The expression of NeoR and IL-2 in different organs was observed in normal syngeneic mice (2 × 10 ~ 6 / mouse). The results showed that NeoR gene-modified hepatocytes were transplanted in the spleen after 24 hours, NeoR gene mRNA was detected in the liver for more than 11 weeks. In addition, the NeoR gene was transiently expressed in the spleen (24h to 1 week) and transiently in the lung tissue (48 to 96 hours) After IL-2 gene-modified hepatocytes, stable expression of IL-2 mRNA (24h to 11 weeks) and IL-2 in peripheral blood (5-40pg / mL) could be detected in the liver, which can enhance hepatic Kupffer Cell Ia antigen expression and NK cytotoxic activity of splenocytes.It is suggested that genetically modified hepatocytes can be targeted to the liver after being transplanted into the liver and assimilated into the liver tissue for long term survival and effective expression of foreign genes Possible ways of gene therapy.