Department of Internal Medicine, Nephrology and Transplantation, Erasmus Medical Center Transplant Institute, University Medical Center Rotterdam, Rotterdam, the Netherlands.
Division of Hepato-pancreatobiliary and Transplant Surgery, Department of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, Rotterdam, the Netherlands.
Transplantation. 2024 Jul 1;108(7):1551-1557. doi: 10.1097/TP.0000000000004936. Epub 2024 Apr 1.
Machine perfusion is the preferred preservation method for deceased donor kidneys. Perfusate fluid, which contains a complex mixture of components, offers potential insight into the organ's viability and function. This study explored immune cell release, particularly tissue-resident lymphocytes (TRLs), during donor kidney machine perfusion and its correlation with injury markers.
Perfusate samples from hypothermic machine perfusion (HMP; n = 26) and normothermic machine perfusion (NMP; n = 16) of human donor kidneys were analyzed for TRLs using flow cytometry. Residency was defined by expressions of CD69, CD103, and CD49as. TRL release was quantified exclusively in NMP. Additionally, levels of cell-free DNA, neutrophil gelatinase-associated lipocalin, and soluble E-cadherin (sE-cadherin) were measured in NMP supernatants with quantitative polymerase chain reaction and enzyme-linked immunosorbent assay.
Both HMP and NMP samples contained a heterogeneous population of TRLs, including CD4 + tissue-resident memory T cells, CD8 + tissue-resident memory T cells, tissue-resident natural killer cells, tissue-resident natural killer T cells, and helper-like innate lymphoid cells. Median TRL proportions among total CD45 + lymphocytes were 0.89% (NMP) and 0.84% (HMP). TRL quantities in NMP did not correlate with donor characteristics, perfusion parameters, posttransplant outcomes, or cell-free DNA and neutrophil gelatinase-associated lipocalin concentrations. However, CD103 + TRL release positively correlated with the release of sE-cadherin, the ligand for the CD103 integrin.
Human donor kidneys release TRLs during both HMP and NMP. The release of CD103 + TRLs was associated with the loss of their ligand sE-cadherin but not with general transplant injury biomarkers.
机器灌注是保存已故供体肾脏的首选方法。灌注液含有复杂的成分混合物，为了解器官的活力和功能提供了潜在的线索。本研究探讨了供体肾脏机器灌注过程中免疫细胞（特别是组织驻留淋巴细胞 [TRL]）的释放及其与损伤标志物的相关性。
使用流式细胞术分析低温机器灌注（HMP；n=26）和常温机器灌注（NMP；n=16）供体肾脏的灌注液样本中的 TRL。通过 CD69、CD103 和 CD49as 的表达来定义驻留。仅在 NMP 中定量 TRL 释放。此外，通过定量聚合酶链反应和酶联免疫吸附试验测量 NMP 上清液中的细胞游离 DNA、中性粒细胞明胶酶相关脂质运载蛋白和可溶性 E-钙黏蛋白（sE-cadherin）水平。
HMP 和 NMP 样本均含有异质性 TRL 群体，包括 CD4+组织驻留记忆 T 细胞、CD8+组织驻留记忆 T 细胞、组织驻留自然杀伤细胞、组织驻留自然杀伤 T 细胞和辅助样固有淋巴细胞。在总 CD45+淋巴细胞中，TRL 的中位数比例为 0.89%（NMP）和 0.84%（HMP）。NMP 中的 TRL 数量与供体特征、灌注参数、移植后结果或细胞游离 DNA 和中性粒细胞明胶酶相关脂质运载蛋白浓度无关。然而，CD103+TRL 的释放与 sE-cadherin 的释放呈正相关，sE-cadherin 是 CD103 整合素的配体。
人类供体肾脏在 HMP 和 NMP 期间释放 TRL。CD103+TRL 的释放与它们的配体 sE-cadherin 的丢失有关，但与一般移植损伤生物标志物无关。
