Adenovirus-mediated gene transfer in the transplant setting. Part III. Variables affecting gene transfer in liver grafts.
Adenovirus-mediated gene transfer in the transplant setting. Part III. Variables affecting gene transfer in liver grafts.
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移植环境中腺病毒介导的基因转移。
DOI：
10.1097/00007890-199503150-00004
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发表时间：
1995
期刊：
影响因子：
6.2
通讯作者：
Shaked,A
中科院分区：
文献类型：
--
作者：
Drazan,KE;Wu,L;Shen,XD;Bullington,D;Jurim,O;Busuttil,RW;Shaked,A
我们建立了一个利用腺病毒载体将基因转移到肝脏移植物的有效系统。本研究的目的是检查在冷保存过程中影响基因转移到大鼠肝移植物的变量。我们的研究结果表明，基因转移效率与载体与肝细胞的比例（感染的多重性[MOI]）和暴露于载体的时间长短直接相关。在10∶1和50∶1的MOIs下，肝细胞转导率分别为25 ~ 30%和100%。然而，较高的MOI与显著的死亡率相关。延长低温保存/暴露时间导致转导率增加（MOI为10:1时为50%）。类似的基因转移效率观察到，当载体稀释在乳酸林格氏或威斯康星大学的解决方案。重组蛋白的产生在再灌注后12小时内明显，并在48小时内达到峰值。这些结果表明，在腺病毒载体体外转导肝移植物后，基因转移和表达的可预测模式。这些数据对于指导移植器官内重组蛋白的理想水平至关重要。
We have established a system of efficient gene transfer to liver grafts using adenovirus vectors. The purpose of this study was to examine variables affecting gene transfer to rat liver grafts during cold preservation. Our results demonstrate that gene transfer efficiency was directly correlated with the ratio of vector to hepatic cells (multiplicity of infection [MOI]) and the length of exposure to the vector. At MOIs of 10: 1 and 50: 1, the hepatic cell transduction rate was 25–30% and 100%, respectively. However, higher MOI was associated with significant mortality. Prolonging the cold preservation/exposure time resulted in an increased transduction rate (50% at MOI of 10: 1). Similar gene transfer efficiencies were observed when the vector was diluted in lactated Ringer's or University of Wisconsin solution. Recombinant protein production was evident within 12 hr after reperfusion, and increased to a peak level within 48 hr. These results suggest a predictable pattern of gene transfer and expression after ex vivo transduction of liver grafts with adenovirus vectors. These data are essential in directing desirable levels of recombinant protein within the transplanted organ.