Division of Thoracic Surgery, Department of Surgery, Mannheim University Medical Center, Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68135 Mannheim, Germany.
Eur J Cardiothorac Surg. 2010 Apr;37(4):859-63. doi: 10.1016/j.ejcts.2009.10.029. Epub 2009 Nov 30.
Alleviation of oxidative stress via targeted delivery of catalase to the pulmonary endothelium by conjugation of angiotensin-converting-enzyme (ACE) monoclonal antibodies attenuates lung injury in an in vivo model of warm lung ischaemia and reperfusion. This study evaluates treatment of lung allografts with conjugates of anti-ACE antibody with catalase (9B9-CAT) in the setting of hypothermic preservation and reports the effect on ischaemia/reperfusion injury in this model.
Rats were injected 1h prior to lung harvesting with mouse immunoglobulin G (IgG) (negative controls), catalase only (CAT) or anti-ACE mAb 9B9 conjugated with catalase (9B9-CAT). Lungs were flushed with low-potassium dextran (LPD) solution, excised and stored at 4 degrees C for 4 and 8h. Grafts were isolated and directly reperfused at 37 degrees C for up to 180 min. Peak inspiratory pressure (PIP), pulmonary arterial pressure (PAP) and lung weight were measured during reperfusion. Anti-oxidative capacity and catalase activity were measured in frozen lung tissue and inflammatory parameters were detected during reperfusion in perfusate solution.
Cold ischaemia of 8h significantly increased lung weight gain, PIP and PAP in non-immune mouse IgG and CAT-treated lungs than in 9B9-CAT-treated lungs (p<0.005). Significantly higher catalase activity and anti-oxidative status were found in the lung tissue of animals conditioned with 9B9-CAT after 4 and 8h of cold storage than in animals treated with catalase (CAT) alone or in animals treated with non-immune mouse IgG (p<0.01).
These results validate immunotargeting by anti-ACE mAb conjugated with catalase as a prospective and specific strategy to augment anti-oxidative defence of the pulmonary endothelium during lung transplantation. Vascular immunotargeting of anti-oxidative enzymes could limit reactive oxygen species mediated ischaemia-reperfusion (I/R) injury of the lung and has the potential to become a promising modality for extension of the viability of banked transplantation tissue.
通过将血管紧张素转换酶（ACE）单克隆抗体与过氧化氢酶偶联，靶向递送至肺内皮细胞，减轻氧化应激，可减轻体内温肺缺血再灌注模型中的肺损伤。本研究评估了在低温保存条件下用抗 ACE 抗体与过氧化氢酶偶联物（9B9-CAT）处理肺移植物，并报告了该方法对该模型中缺血/再灌注损伤的影响。
在肺采集前 1 小时，向大鼠注射小鼠免疫球蛋白 G（IgG）（阴性对照）、过氧化氢酶（CAT）或与过氧化氢酶偶联的抗 ACE mAb 9B9（9B9-CAT）。用低钾右旋糖酐（LPD）溶液冲洗肺，切除并在 4°C 下保存 4 和 8 小时。将移植物分离并在 37°C 下直接再灌注长达 180 分钟。在再灌注过程中测量吸气峰压（PIP）、肺动脉压（PAP）和肺重量。在冷冻肺组织中测量抗氧化能力和过氧化氢酶活性，并在再灌注过程中检测灌流液中的炎症参数。
8 小时的冷缺血显著增加了非免疫小鼠 IgG 和 CAT 处理的肺中的肺重量增加、PIP 和 PAP，而 9B9-CAT 处理的肺则没有（p<0.005）。在冷储存 4 和 8 小时后，用 9B9-CAT 处理的动物的肺组织中发现了更高的过氧化氢酶活性和抗氧化状态，而单独用 CAT 处理的动物或用非免疫小鼠 IgG 处理的动物则没有（p<0.01）。
这些结果验证了用抗 ACE mAb 与过氧化氢酶偶联的免疫靶向作为一种有前途的、特异性的策略，可增强肺移植过程中肺内皮细胞的抗氧化防御。血管免疫靶向抗氧化酶可限制活性氧介导的肺缺血再灌注（I/R）损伤，并有潜力成为延长储存移植组织活力的有前途的方法。
