小鼠后肢缺血模型中新型“假毒粒”型重组血清型1型腺相关病毒载体的安全性研究  

The safety assessment of gene transfer mediated by the new recombinant adeno-associated virus se- rotype 1 pseudotyped vector in mouse ischemic lamb model

作　　者：鄢华[1] 吴小兵[2] 郭艳红[1] 高炜[1] 

机构地区：[1]北京大学第三医院心内科,100083 [2]中国疾病预防控制中心病毒病所国家重点实验室

出　　处：《中华实验外科杂志》2013年第7期1497-1499,共3页Chinese Journal of Experimental Surgery

基　　金：国家高技术研究发展计划资助项目（2001AA217111、2002AA223324）

摘　　要：目的探讨骨骼肌注射法导入新型“假毒粒”型重组血清1型腺相关病毒（rAAVl）载体的安全性。方法构建小鼠后肢缺血模型，术后10d每只小鼠股部缺血骨骼肌内注射3×10^11vg的rAAV1-LacZ、rAAV1-VEGF165载体，检测载体介导外源基因表达的高峰时间和持续时间；观察小鼠血循环、各主要脏器中有无外源基因表达。结果载体转染后1月外源基因表达至高峰，表达持续时间超过9个月。rAAV1-VEGF165载体转染后1月酶联免疫吸附试验（ELISA）法检测小鼠血浆样品未发现VEGF螂表达。rAAVI—LacZ载体转染后1个月小鼠各主要脏器病理检测均未发现外源LacZ基因表达。结论骨骼肌注射法导入新型“假毒粒”型rAAVl载体安全，rAAV1-VEGF165载体是治疗外周动脉疾病的1种安全载体。Objective To investigate the safety of the new recombinant adeno-associated virus se- rotype 1 （rAAV1） pseudotyped vector medicating gene transfer into mouse ischemic skeletal muscles. Methods Ten days following ischemia induction in mouse hind limbs, 3 ×10^11 vg per mouse rAAV1-LacZ or rAAV1-VEGF165 vectors were injected in ischemic thigh muscles. Gene transfer efficiency and gene ex- pression duration were observed. Heterologous gene expression was tested in mouse plasma samples and mouse important organs. Results Gene expression achieved the highest level at 1 month after gene trans- fer and lasted more than 9 months. At 1 month after gene transfer, no VEGF165 protein was found in mouse plasma by enzyme linked immunosorbent assay （ELISA）. No heterologous LacZ gene expression was de- tectable in mouse organs. Conclusion Vector injection into the skeletal muscle is a safe gene transfer method for the new rAAV1 pseudotyped vector. The rAAV1-VEGF165 vector may be a superior promising vector mediating gene therapy for peripheral vascular diseases.

关 键 词：腺相关病毒 血清型 载体 基因治疗 

分 类 号：R-332[医药卫生] R5