The efficacy and safety of gene transfer into the porcine liver in vivo by HVJ (Sendai virus) liposome
The efficacy and safety of gene transfer into the porcine liver in vivo by HVJ (Sendai virus) liposome
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DOI：
10.1097/01.tp.0000184447.88283.f3
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发表时间：
2005-12-15
期刊：
影响因子：
6.2
通讯作者：
Kanematsu, T
中科院分区：
文献类型：
--
作者：
Kawashita, Y;Fujioka, H;Kanematsu, T
背景使用病毒载体的基因转移系统是有效的;然而，大多数病毒载体也倾向于引起免疫反应，从而在临床上引起一系列副作用。HVJ-脂质体载体是由脂质体和灭活仙台病毒（日本血友病病毒[HVJ]）组成的混合载体，据报道其免疫原性较低，也可以重复给药。我们在猪模型中检查了该载体用于肝脏基因治疗的有用性。将编码β-半乳糖苷酶和荧光素酶的基因用作报告基因。在完全阻断肝脏血管的情况下，向猪的门静脉中注射负载报告基因的-HV]-脂质体。然后通过β-半乳糖苷酶染色、荧光素酶测定和LacZ mRNA的RT-PCR评估转染效率。进行生化和组织学分析以评估基因转移后的组织毒性。转染后第7天肝脏中荧光素酶基因表达达到最高水平。在整个观察期内，在转染后28天内继续检测到，而所有猪保持健康。根据β-半乳糖苷酶染色，肝细胞中的转染效率为15%。肝外转基因表达在肺和肾中有少量表达，但在脾和卵巢中没有表达。这些数据首次表明，HVJ-脂质体载体的使用是一种安全可行的猪肝脏定向基因转移方式，因此可能适用于临床基因治疗试验。
Background. Gene transfer systems using viral vectors are efficient; however, most viral vectors also tend to evoke immunologic reactions, thereby clinically causing serial side effects. HVJ-liposome vector is a hybrid vector consisting of liposome and an inactivated Sendai virus (Hemmagglutinating Virus of Japan [HVJ]), which has been reported to be less immunogenic and can also be repeatedly administered. We examined the usefulness of this vector for hepatic gene therapy in a pig model.Methods. Genes encoding beta-galactosidase and luciferase were used as reporter genes. The pigs were injected with the reporter gene loaded-HV]-liposome into the portal vein under total vascular exclusion of the liver. The transfection efficiencies were then assessed by beta-galactosidase staining, a luciferase assay, and RT-PCR for LacZ mRNA. Biochemical and histologic analyses were performed to evaluate tissue toxicity after gene transfer.Results. The luciferase gene expression in the liver reached its highest level at 7 days after transfection. It continued to be detected up to 28 days after transfection, while all pigs remained health), throughout the observation period. The transfection efficiency was 15% in the hepatocytes according to beta-galactosidase staining. Extrahepatic transgene expression was slightly observed in the lung and kidney, but not in the spleen or ovary.Conclusions. These data suggest for the first time that the use of the HVJ-liposome vector is a safe and feasible modality for liver-directed gene transfer in pigs, and it might therefore be suitable for clinical gene therapy trials.