Expression of the hepatitis C virus genome in rat liver after cationic liposome‐mediated in vivo gene transfer
Expression of the hepatitis C virus genome in rat liver after cationic liposome‐mediated in vivo gene transfer
复制标题
阳离子脂质体介导的体内基因转移后丙型肝炎病毒基因组在大鼠肝脏中的表达
作者：
T. Takehara;N. Hayashi;Y. Miyamoto;Masato Yamamoto;E. Mita;H. Fusamoto;T. Kamada
由于缺乏丙型肝炎病毒（HCV）感染的小动物模型，阻碍了阐明HCV的发病机制。本研究的目的是通过阳离子脂质体介导的体内基因转移建立HCV表达动物模型。为了检查这种策略的可行性，将pActLacZ（一种由β-肌动蛋白启动子驱动的LacZ基因组成的表达载体，与脂质体复合）逆行注射到成年大鼠的胆总管中。X-Gal组织化学染色清楚地显示LacZ基因在肝细胞中表达，但在胆管上皮细胞中不表达。在1：4的DNA与脂质体比率下观察到最大表达。基于这一观察结果，评价了pAGS 3 M091，一种含有全长HCV互补DNA（cDNA）的表达载体，前接β-肌动蛋白启动子。体内胆管内给予与脂质体复合的pAGS 3 M091后2天，逆转录的肝脏RNA的聚合酶链反应（PCR）扩增证实了来自pAGS 3 M091的HCV转录物的5′和3′部分。免疫组化分析显示HCV核心蛋白在少量肝细胞内散在分布于肝小叶内。我们的结论是，全长HCV基因组成功地表达在成年大鼠肝脏通过阳离子脂质体介导的体内基因转移。该模型为进一步研究HCV在体内的免疫病理学作用提供了一个新的模型。
The lack of a small animal model of hepatitis C virus (HCV) infection has impeded elucidation of the pathogenesis of HCV. The aim of this study was to develop an HCV‐expressing animal model by means of cationic liposome‐mediated in vivo gene transfer. To examine the feasibility of this strategy, pActLacZ, an expression vector composed of the LacZ gene driven by the β‐actin promoter, complexed with lipofectin, was injected retrogradely into the common bile ducts of adult rats. X‐Gal histochemical staining clearly showed that the LacZ gene was expressed in hepatocytes, but not in biliary epithelial cells. Maximal expression was observed at a DNA to lipofectin ratio of 1:4. Based on this observation, pAGS3M091, an expression vector containing the full length of HCV complementary DNA (cDNA) preceded by the β‐actin promoter, was evaluated. Two days after in vivo intrabiliary administration of pAGS3M091 complexed with lipofectin, polymerase chain reaction (PCR) amplification of reverse‐transcribed liver RNA demonstrated the 5′ and 3′ portions of HCV transcripts derived from pAGS3M091. Immunohistochemical analysis showed the HCV core protein in a small number of hepatocytes scattered in the hepatic lobules. We conclude that the full‐length HCV genome was successfully expressed in adult rat liver by means of cationic liposome‐mediated in vivo gene transfer. This model will be useful for detemining the immunopathological role of HCV in vivo.
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影响因子：
56.9
作者：
E. Nabel;G. Plautz;G. Nabel
通讯作者：
E. Nabel;G. Plautz;G. Nabel
使用含有磷酸烯醇丙酮酸羧激酶基因启动子的逆转录病毒在动物中进行肝基因转移。
DOI：
--
发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hatzoglou,M;Lamers,W;Bosch,F;Wynshaw-Boris,A;Clapp,DW;Hanson,RW
通讯作者：
Hanson,RW
将体内基因直接转移到完整狗的冠状动脉和外周脉管系统中。
影响因子：
37.8
作者：
Lim,CS;Chapman,GD;Gammon,RS;Muhlestein,JB;Bauman,RP;Stack,RS;Swain,JL
通讯作者：
Swain,JL
逆转录病毒介导的外源基因体内局部递送至大鼠肝细胞：需要部分肝切除才能成功表达外源基因。
影响因子：
3.8
作者：
Moscioni,AD;Rozga,J;Neuzil,DF;Overell,RW;Holt,JT;Demetriou,AA
通讯作者：
Demetriou,AA
兔模型中的经皮动脉基因转移。
DOI：
10.1172/jci115970
发表时间：
1992
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Leclerc,G;Gal,D;Takeshita,S;Nikol,S;Weir,L;Isner,JM
通讯作者：
Isner,JM