Drazan K E, Wu L, Shen X D, Bullington D, Jurim O, Busuttil R W, Shaked A
Department of Surgery, University of California at Los Angeles.
Transplantation. 1995 Mar 15;59(5):670-3. doi: 10.1097/00007890-199503150-00004.
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.
我们已经建立了一种使用腺病毒载体向肝移植供体高效转移基因的系统。本研究的目的是检测在冷保存期间影响大鼠肝移植供体基因转移的变量。我们的结果表明，基因转移效率与载体与肝细胞的比例（感染复数[MOI]）以及暴露于载体的时间直接相关。在MOI为10:1和50:1时，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。延长冷保存/暴露时间会导致转导率增加（在MOI为10:1时为50%）。当载体在乳酸林格氏液或威斯康星大学溶液中稀释时，观察到相似的基因转移效率。再灌注后12小时内即可明显检测到重组蛋白的产生，并在48小时内增加至峰值水平。这些结果表明，用腺病毒载体对肝移植供体进行体外转导后，基因转移和表达具有可预测的模式。这些数据对于指导移植器官内重组蛋白达到理想水平至关重要。
