The Ex-Vivo Lab, Division of Cell Matrix and Regenerative Medicine, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, School of Biological Sciences, The University of Manchester, Manchester, United Kingdom.
The Transplant Centre, Manchester University Hospitals NHS Foundation Trust, Manchester, United Kingdom.
Front Immunol. 2020 Jul 28;11:1621. doi: 10.3389/fimmu.2020.01621. eCollection 2020.
Many donor organs contain significant leukocyte reservoirs which upon transplantation activate recipient leukocytes to initiate acute rejection. We aimed to assess whether non-ischemic heart preservation via perfusion promotes immunodepletion and alters the inflammatory status of the donor organ prior to transplantation. Isolated porcine hearts underwent hypothermic, cardioplegic perfusion for 8 h. Leukocyte populations were quantified in left ventricle samples by flow cytometry. Cell-free DNA, cytokines, and chemokines were quantified in the perfusate. Tissue integrity was profiled by targeted proteomics and a histological assessment was performed. Heterotopic transplants comparing hypothermic preservation and static cold storage were utilized to assess graft infiltration as a solid clinical endpoint. perfusion significantly immunodepleted myocardial tissue. The perfusate displayed a selective, pro-inflammatory cytokine/chemokine pattern dominated by IFN-γ. The tissue molecular profile was improved following perfusion by diminished expression of nine pro-apoptotic and six ischemia-associated proteins. Histologically, no evidence of tissue damage was observed and cardiac troponin I was low throughout perfusion. Cell-free DNA was detected, the source of which may be necrotic/apoptotic leukocytes. Post-transplant graft infiltration was markedly reduced in terms of both leucocyte distribution and intensity of foci. These findings demonstrate that perfusion significantly reduced donor heart immunogenicity via loss of resident leukocytes. Despite the pro-inflammatory cytokine pattern observed, a pro-survival and reduced ischemia-related profile was observed, indicating an improvement in graft viability by perfusion. Diminished graft infiltration was observed in perfused hearts compared with those preserved by static cold storage following 48 h of transplantation.
许多供体器官含有大量白细胞储备，这些白细胞在移植后会激活受体白细胞，引发急性排斥反应。我们旨在评估通过灌注进行非缺血性心脏保存是否可以促进免疫耗竭，并改变供体器官在移植前的炎症状态。 离体猪心在低温、心脏停搏液下灌注 8 小时。通过流式细胞术对左心室样本中的白细胞群进行定量。在灌流液中定量细胞游离 DNA、细胞因子和趋化因子。通过靶向蛋白质组学分析组织完整性，并进行组织学评估。比较低温保存和静态冷藏的异位移植用于评估移植物浸润作为实体临床终点。 灌注显著地免疫耗竭了心肌组织。灌流液显示出一种选择性的、促炎细胞因子/趋化因子模式，主要由 IFN-γ 主导。灌注后，组织分子谱通过减少表达 9 种促凋亡和 6 种与缺血相关的蛋白质得到改善。组织学上，没有观察到组织损伤的证据，整个灌注过程中心肌钙蛋白 I 水平较低。检测到细胞游离 DNA，其来源可能是坏死/凋亡的白细胞。与白细胞分布和焦点强度相比，移植后移植物浸润明显减少。 这些发现表明，通过丢失驻留白细胞，灌注显著降低了供体心脏的免疫原性。尽管观察到促炎细胞因子模式，但观察到促生存和减少与缺血相关的谱，表明灌注可改善移植物活力。与静态冷藏保存的心脏相比，灌注保存的心脏在移植后 48 小时的移植物浸润明显减少。
