Boasquevisque C H, Mora B N, Boglione M, Ritter J K, Scheule R K, Yew N, Debruyne L, Qin L, Bromberg J S, Patterson G A
Division of Cardiothoracic Surgery, Department of Surgery, and Department of Pathology, Washington University School of Medicine, St. Louis, MO, USA.
J Thorac Cardiovasc Surg. 1999 Jan;117(1):8-14; discussion 14-5. doi: 10.1016/s0022-5223(99)70463-0.
We compared the efficacy of in vivo and ex vivo liposome transfection in rat lung transplantation.
(1) Chloramphenicol acetyltransferase group: Fischer rats underwent isogeneic transplantation (n = 4 per group). Recipients were put to death on postoperative day 2 for chloramphenicol acetyltransferase activity. Ex vivo setting: Grafts received cDNA complexed or not with liposomes and were transplanted after 1.5 or 10 hours at 10 degreesC. In vivo setting: Donors were intravenously injected with cDNA complexed or not with liposomes. Lungs were harvested after 1.5 or 10 hours, preserved at 10 degreesC, and transplanted. (2) Transforming growth factor-beta1 group: Brown-Norway rats served as donors and Fischer rats as recipients. All grafts were preserved for 3 hours at 10 degreesC. On postoperative day 5, arterial oxygenation and histologic rejection scores were assessed. Ex vivo setting: Grafts received transforming growth factor-beta1 sense (n = 8) or antisense (n = 7) complexed with liposomes or cDNA alone (n = 5). In vivo setting: Donors were intravenously injected with liposome:transforming growth factor-beta1 sense cDNA (n = 7). Exposure time was 3 hours.
(1) Chloramphenicol acetyltransferase-transfection was superior in the ex vivo group but was not statistically different for longer exposure times. (2) Transforming growth factor-beta1-arterial oxygenation was superior in the ex vivo liposome:sense group. cDNA alone was inefficient. Rejection scores were not statistically different between ex vivo and in vivo liposome:sense groups but were better when the ex vivo liposome:sense group was compared with the cDNA alone or the antisense groups.
(1) With current liposome technology, the ex vivo route is superior to the in vivo approach; (2) cDNA alone does not provide transgene expression at levels to produce a functional effect.
我们比较了体内和体外脂质体转染在大鼠肺移植中的效果。
（1）氯霉素乙酰转移酶组：Fischer大鼠接受同基因移植（每组n = 4）。术后第2天处死受体，检测氯霉素乙酰转移酶活性。体外设置：移植物接受与脂质体复合或未复合的cDNA，并在10℃下1.5或10小时后移植。体内设置：供体静脉注射与脂质体复合或未复合的cDNA。1.5或10小时后收获肺，在10℃下保存，然后移植。（2）转化生长因子-β1组：以Brown-Norway大鼠为供体，Fischer大鼠为受体。所有移植物在10℃下保存3小时。术后第5天，评估动脉氧合和组织学排斥评分。体外设置：移植物接受与脂质体复合的转化生长因子-β1正义链（n = 8）或反义链（n = 7）或单独的cDNA（n = 5）。体内设置：供体静脉注射脂质体：转化生长因子-β1正义链cDNA（n = 7）。暴露时间为3小时。
（1）氯霉素乙酰转移酶转染在体外组中更优，但暴露时间较长时无统计学差异。（2）转化生长因子-β1动脉氧合在体外脂质体：正义链组中更优。单独的cDNA效率低下。体外和体内脂质体：正义链组之间的排斥评分无统计学差异，但与单独的cDNA或反义链组相比，体外脂质体：正义链组的评分更好。
（1）使用当前的脂质体技术，体外途径优于体内途径；（2）单独的cDNA不能产生具有功能效应水平的转基因表达。
