O043 Evaluation of minimal factor H therapy administered to kidneys during ex vivo normothermic perfusion as a treatment to improve ischaemia reperfusion injury
O043 Evaluation of minimal factor H therapy administered to kidneys during ex vivo normothermic perfusion as a treatment to improve ischaemia reperfusion injury
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O043 离体常温灌注期间对肾脏进行最小因子 H 治疗作为改善缺血再灌注损伤的治疗的评估
DOI：
10.1093/bjs/znad101.043
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发表时间：
2023
影响因子：
9.6
通讯作者：
Connelly C
中科院分区：
文献类型：
--
作者：
Connelly C
补体激活是缺血再灌注损伤的一个重要机制，尤其是旁路途径。旁路途径的主要调节因子是H因子。我们假设，同型二聚体的小因子H（HDM-FH; PMID：29588430）可能会保护移植肾补体介导的损伤时，在常温灌注机perfusion.MethodsKidneys期间，从女性白色长白猪检索优化协议。每对中的一个肾脏随机接受5 mg HDM-FH（108 μg/mL）。在37°C下用自体血液灌注肾脏6小时。使用ELISA和免疫荧光测量HDM-FH结合。通过定量组织中的Bb沉积来测量补体激活。结果HDM-FH在灌流过程中与灌流液的结合量约为4 mg，在尿中的结合量小于10%，提示HDM-FH已达到饱和状态。使用免疫荧光证实肾脏内的HDM-FH结合。HDM-FH定位于肾小球，随着灌注时间的推移沉积增加。接受HDM-FH的肾脏中旁路途径激活减少，如Bb沉积减少所示。纤维蛋白原沉积也减少，并发现与C3在肾小球共定位。HDM-FH抑制补体活性在血清中的剂量依赖manow.ConclusionInfusion与HDM-FH之前，模拟肾移植条件减少补体激活和其他继发性的负面结果的器官。因此，用HDM-FH进行器官灌注极有可能有助于延长移植后的移植物存活，这将在未来的研究中进行评估。
IntroductionComplement activation is a key mechanism of Ischaemia reperfusion injury, with the alternative pathway driving damage in particular. The main regulator of the alternative pathway is factor H. We hypothesised that homodimeric mini-factor H (HDM-FH; PMID:29588430) may protect transplanted kidneys from complement mediated damage when administered during normothermic perfusion machine perfusion.MethodsKidneys were retrieved from female white landrace pigs following an optimised protocol. One kidney from each pair was randomised to receive 5mg of HDM-FH (∼8μg/mL). Kidneys were perfused at 37°C with autologous blood for 6 hours. HDM-FH binding was measured using ELISA and immunofluorescence. Complement activation was measured by quantifying Bb deposition in tissue. The ability of HDM-FH to inhibit complement in serum was measured using haemolytic assays.Results4mgs of HDM-FH bound from perfusate during perfusion, with <10% lost in urine suggesting saturation was achieved. HDM-FH binding within the kidneys was confirmed using immunofluorescence. HDM-FH localised to glomeruli with deposition increasing over the time of the perfusion. Alternative pathway activation was reduced in kidneys receiving HDM-FH as demonstrated by reduced Bb deposition. Fibrinogen deposition was also reduced and found to colocalise with C3 in glomeruli. HDM-FH inhibits complement activity in serum in a dose-dependent manor.ConclusionInfusion with HDM-FH prior to simulated kidney transplant conditions reduced complement activation and other secondary negative outcomes to the organ. Therefore, organ perfusion with HDM-FH is highly likely to help prolong graft survival after transplant and this will be assessed in future studies.