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

本研究旨在探讨在移植过程中转染NF-κ B诱饵基因是否能减少一氧化氮（NO）的产生，减轻急性肺排斥反应。使用成对的Brown Norway（RT 1n）和刘易斯（RT 1I）大鼠进行左肺移植。所有移植物均用含有日本血凝病毒（HVJ）脂质体-NF-κ B诱饵ODNs复合物的PBS溶液20 ml冲洗（n = 6），4 ℃保存60 min。移植后5天，不使用免疫抑制剂，呼出NO，气体交换和移植物的组织学排斥反应评分。与对照组相比，NF-kB诱饵转染组呼出的NO水平显著降低（445 ± 162 vs.1305 ± 123 ppb，p <0.02），PaO2（197 ± 28 vs.60 ± 18 mmHg，p <0.02）和排斥评分（1.6 ± 0.4 vs.2.5 ± 0.4）也有所改善。通过ODNs诱骗转染供体肺来抑制移植物中NF-κ B的活化，减轻了急性移植排斥反应中的肺损伤。这一结果表明，一个可能的治疗靶点，异基因非依赖性途径，这似乎是可行的，在肺移植的临床设置与免疫抑制。

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.