Gene therapy in lung transplantation. Attenuation of ischemia/reperfusion injury and acute rejection by NF-kB decoy transfection

本研究旨在确定在收获期间将核因子-kB诱骗基因导入同种异体移植物是否能减少一氧化氮(NO)的产生，从而改善急性肺排斥反应。左肺移植使用棕色挪威(RT1N)和刘易斯(RT1I)大鼠对。用含日本血凝素(Hvj)脂质体-核因子-kB诱骗寡核苷酸复合体(n=6)的PBS液冲洗移植物，在4℃保存60min，以杂乱诱骗为对照(n=5)和单纯PBS液(n=5)。移植后第5天，不使用免疫抑制剂，测定移植肾呼出的NO、气体交换和组织学排斥反应评分。与对照组相比，NF-kB诱骗转染组呼出的NO水平显著降低(445±162vs.1305±123 ppb，p&lt；0.02)，PaO2(197±28vs.60±18 mmHg，p&lt；0.02)和排斥评分(1.6±0.4vs.2.5±0.4)明显改善。ODNS诱骗转染法抑制同种异体肺移植中核因子-kB的活化减轻急性排斥反应中肺损伤。这一结果暗示了异体非依赖途径的一个可能的治疗靶点，结合免疫抑制在肺移植的临床环境中似乎是可行的。

This study are to determine whether NF-kB decoy transfection into the allograft during harvest reduce nitric oxide (NO) production and ameliorate acute lung rejection. Left lung transplantation was performed using pairs of Brown Norway(RT1n) and Lewis (RT1I) rats. All allografts were flushed with PBS solution 20 ml containing hemaglutinating virus of Japan (HVJ) liposome-ODNs complex of NF-kB decoy (n=6) and preserved for 60 minutes at 4℃, Scrambled decoy was regarded as control (n=5) as well as simple PBS solution (n=5). Five days after transplantation without use of immunosuppressants, exhaled NO, gas exchange and histological rejection score of the graft were determined. Exhaled NO level significantly reduced in group with NF-kB decoy transfection compared with control (445±162 vs. 1305±123 ppb, p<0.02) as well as improvement in PaO2 (197±28 vs. 60±18 mmHg, p<0.02) and rejection score (1.6±0.4 vs. 2.5±0.4). Inhibition of NF-kB activation in the allograft by ODNs decoy transfection into the donor lung ameliorated lung injury in acute allograft rejection. This result implies a possible therapeutic target for allo-independent pathway, which seemed to be feasible in conjunction with immuinosuppression in the clinical setting of lung transplantation.