D'Ovidio F, Yano M, Ritter J H, Mohanakumar T, Patterson G A
Department of Surgery, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Ann Thorac Surg. 1999 Sep;68(3):1008-13. doi: 10.1016/s0003-4975(99)00784-5.
We investigated endobronchial transfection of CAT and TGF-beta1 cDNA selectively delivered to the lung graft with or without liposomes.
Phase I: F344 rats received 130 microg of naked plasmid pCF1-CAT or complexed to liposome GL67 via left main bronchus instillation. Rats were awakened (pCF1-CAT, n = 4; GL67:pCF1-CAT, n = 4) or served as donors in an isogenic transplant (pCF1-CAT, n = 5; GL67:pCF1-CAT, n = 5). ELISA was performed on lungs, hearts, and livers on POD 2. Phase II: BN lungs received TGF-beta1 sense (n = 6); antisense (n = 5); GL67:TGF-beta1 sense (n = 10); or saline solution (n = 10). F344 recipients were sacrificed on POD 5. The arterial pO2 and rejection were assessed. RT-PCR for murine TGF-beta1 was performed.
Phase I: CAT expression was 519+/-287 pg and 63+/-68 with pCF1-CAT and 104+/-67 and 37+/-45 with GL67:pCF1-CAT, respectively, in the non-transplant and in the transplant setting. No protein was detected in the hearts, livers, and in the native lung of the recipients. Phase II: RT-PCR confirmed murine TGF-beta1 transfection. pO2 was 362.7+/-110.2 (mean mm Hg +/- SD) for sense TGF-beta1; 146.88+/-85.5 for antisense; 241.5+/-181.5 for GL67-TGF-beta1 sense; and 88.4+/-38.7 for saline. TGF-beta1 sense versus all other groups, p<0.05, GL67-TGF-beta1 sense versus saline, p = 0.01. Rejection was significantly lower for TGF-beta1 sense versus saline, p = 0.04.
Endobronchial administration of naked plasmid achieves selective transfection of lung grafts. Using this strategy, TGF-beta1 reduces early lung allograft rejection.
我们研究了通过支气管内转染将氯霉素乙酰转移酶（CAT）和转化生长因子-β1（TGF-β1）的互补脱氧核糖核酸（cDNA）选择性递送至肺移植物，转染过程中使用或不使用脂质体。
第一阶段：F344大鼠通过左主支气管滴注接受130微克裸质粒pCF1-CAT或与脂质体GL67复合的pCF1-CAT。大鼠苏醒（pCF1-CAT组，n = 4；GL67:pCF1-CAT组，n = 4）或作为同基因移植的供体（pCF1-CAT组，n = 5；GL67:pCF1-CAT组，n = 5）。在术后第2天对肺、心脏和肝脏进行酶联免疫吸附测定（ELISA）。第二阶段：BN大鼠的肺接受TGF-β1正义链（n = 6）；反义链（n = 5）；GL67:TGF-β1正义链（n = 10）；或生理盐水（n = 10）。F344受体在术后第5天处死。评估动脉血氧分压（pO2）和排斥反应。进行小鼠TGF-β1的逆转录聚合酶链反应（RT-PCR）。
第一阶段：在非移植和移植情况下，pCF1-CAT组肺组织中CAT表达分别为519±287皮克和63±68皮克，GL67:pCF1-CAT组分别为104±67皮克和37±45皮克。在受体的心脏、肝脏和原生肺中未检测到蛋白质。第二阶段：RT-PCR证实了小鼠TGF-β1的转染。TGF-β1正义链组pO2为362.7±110.2（平均毫米汞柱±标准差）；反义链组为146.88±85.5；GL67-TGF-β1正义链组为241.5±181.5；生理盐水组为88.4±38.7。TGF-β1正义链组与所有其他组相比，p<0.05；GL67-TGF-β1正义链组与生理盐水组相比，p = 0.01。TGF-β1正义链组与生理盐水组相比，排斥反应明显更低，p = 0.04。
支气管内给予裸质粒可实现肺移植物的选择性转染。采用这种策略，TGF-β1可降低早期肺移植排斥反应。
