为实施临床ABO血型不相容肝细胞移植而进行的人分离肝细胞血型抗原表达研究。
Blood Group Antigen Expression in Isolated Human Liver Cells in Preparation for Implementing Clinical ABO-Incompatible Hepatocyte Transplantation.
作者信息
Tokodai Kazuaki, Kumagai-Braesch Makiko, Karadagi Ahmad, Johansson Helene, Ågren Nils, Jorns Carl, Ericzon Bo-Göran, Ellis Ewa
机构信息
Department of Clinical Science, Intervention and Technology (CLINTEC), Division of Transplantation Surgery, Karolinska Institute and Karolinska University Hospital Huddinge, Stockholm, SE-141 86, Sweden.
出版信息
J Clin Exp Hepatol. 2020 Mar-Apr;10(2):106-113. doi: 10.1016/j.jceh.2019.07.001. Epub 2019 Jul 11.
BACKGROUND
ABO blood group antigens in the liver are expressed mainly on endothelial cells or biliary epithelial cells but not on hepatocytes. This suggests that ABO-incompatible hepatocyte transplantation (ABOi-HTx) is theoretically feasible. However, the effects of stress on ABO blood group antigen expression caused by isolation and intraportal infusion require thorough investigation before ABOi-HTx can be implemented in clinical settings.
METHODS
Human hepatocytes were isolated from liver tissue obtained from liver resection or deceased donor livers. The expression of blood group antigens on cryopreserved human liver tissues and isolated hepatocyte smear specimens were examined by immunofluorescent staining. The effect of proinflammatory cytokines on blood group antigen expression of hepatocytes was evaluated by flow cytometry. Instant blood-mediated inflammatory reaction after hepatocyte incubation with ABO-incompatible whole blood was examined using the tubing loop model.
RESULTS
Blood group antigens were mainly expressed on vessels in the portal area. In hepatocyte smear specimens, isolated hepatocytes did not express blood group antigens. In contrast, a subset of cells in the smear specimens of nonparenchymal liver cells stained positive. In the flow cytometry analysis, isolated hepatocytes were negative for blood group antigens, even after 4-h incubation with cytokines. Platelet counts and complement activation were not significantly different in ABO-identical versus ABO-incompatible settings in the tubing loop model.
CONCLUSION
Our study showed that blood group antigens were not expressed on hepatocytes, even after isolation procedures or subsequent incubation with cytokines. This finding is an important step toward removing the restriction of ABO matching in hepatocyte transplantation. Our results suggest that ABOi-HTx is a feasible therapeutic option, especially in patients who require urgent treatment with freshly isolated hepatocytes, such as those with acute liver failure.
背景
肝脏中的ABO血型抗原主要在内皮细胞或胆管上皮细胞上表达，而不在肝细胞上表达。这表明ABO血型不相容的肝细胞移植（ABOi-HTx）在理论上是可行的。然而，在临床环境中实施ABOi-HTx之前，需要对分离和门静脉输注引起的应激对ABO血型抗原表达的影响进行深入研究。
方法
从肝切除或已故供体肝脏获得的肝组织中分离人肝细胞。通过免疫荧光染色检查冷冻保存的人肝组织和分离的肝细胞涂片标本上血型抗原的表达。通过流式细胞术评估促炎细胞因子对肝细胞血型抗原表达的影响。使用管路环模型检查肝细胞与ABO血型不相容的全血孵育后的即时血液介导的炎症反应。
结果
血型抗原主要在门区血管上表达。在肝细胞涂片标本中，分离的肝细胞不表达血型抗原。相比之下，非实质肝细胞涂片标本中的一部分细胞染色呈阳性。在流式细胞术分析中，即使与细胞因子孵育4小时后，分离的肝细胞血型抗原仍为阴性。在管路环模型中，ABO血型相同与ABO血型不相容的情况下，血小板计数和补体激活没有显著差异。
结论
我们的研究表明，即使经过分离程序或随后与细胞因子孵育，肝细胞上也不表达血型抗原。这一发现是消除肝细胞移植中ABO血型匹配限制的重要一步。我们的结果表明，ABOi-HTx是一种可行的治疗选择，特别是对于那些需要用新鲜分离的肝细胞进行紧急治疗的患者，如急性肝衰竭患者。
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