van de Kerkhove Maarten-Paul, Germans Menno R, Deurholt Tanja, Hoekstra Ruurdtje, Joziasse David H, van Wijk Albert C W A, van Gulik Thomas M, Chamuleau Robert A F M, Roos Anja
Dept. of Surgery (Surgical Laboratory), Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands.
J Hepatol. 2005 Apr;42(4):541-7. doi: 10.1016/j.jhep.2004.11.041. Epub 2005 Jan 22.
BACKGROUND/AIMS: To bridge acute liver failure (ALF) patients to orthotopic liver transplantation, several bioartificial liver (BAL) systems have been developed. The bio-component of most BAL systems consists mainly of porcine hepatocytes. Plasma or blood of ALF patients is perfused through the BAL thereby contacting porcine hepatocytes. Xenogeneic BAL systems may suffer from hyperacute rejection similar to whole-organ xenotransplants. Hyperacute rejection is mediated by antibodies directed against Galalpha(1-3)Gal, a carbohydrate structure present on most mammalian cells. Galalpha(1-3)Gal is produced by the enzyme alpha1,3-galactosyltansferase (alphaGal-T). Conflicting data have been published concerning Galalpha(1-3)Gal expression on hepatocytes in intact porcine liver. We investigated whether isolated porcine hepatocytes express Galalpha(1-3)Gal.
Immunofluorescence, flow cytometry, RT-PCR and enzyme activity assays were performed on freshly isolated and cultured porcine hepatocytes and liver biopsies. Anti-Galalpha(1-3)Gal antibodies were measured in plasma from patients treated with BAL by ELISA.
Isolated porcine hepatocytes express (alphaGal-T) at low levels and Galalpha(1-3)Gal is present in low quantities on these cells, in contrast to hepatocytes in situ. Furthermore, IgG and IgM anti-Galalpha(1-3)Gal are depleted from the plasma of ALF patients during BAL treatment.
Isolation and culture of porcine hepatocytes induce Galalpha(1-3)Gal expression, which may elicit immunological responses potentially compromising BAL functionality.
背景/目的：为了将急性肝衰竭（ALF）患者与原位肝移植相衔接，已开发出多种生物人工肝（BAL）系统。大多数BAL系统的生物成分主要由猪肝细胞组成。ALF患者的血浆或血液通过BAL进行灌注，从而与猪肝细胞接触。异种BAL系统可能会遭受与全器官异种移植类似的超急性排斥反应。超急性排斥反应由针对Galα(1-3)Gal的抗体介导，Galα(1-3)Gal是大多数哺乳动物细胞上存在的一种碳水化合物结构。Galα(1-3)Gal由α1,3-半乳糖基转移酶（αGal-T）产生。关于完整猪肝中肝细胞上Galα(1-3)Gal的表达，已发表了相互矛盾的数据。我们研究了分离的猪肝细胞是否表达Galα(1-3)Gal。
对新鲜分离和培养的猪肝细胞及肝活检组织进行免疫荧光、流式细胞术、逆转录-聚合酶链反应（RT-PCR）和酶活性测定。通过酶联免疫吸附测定（ELISA）检测接受BAL治疗患者血浆中的抗Galα(1-3)Gal抗体。
与原位肝细胞相比，分离的猪肝细胞αGal-T表达水平较低，且这些细胞上Galα(1-3)Gal的含量也较少。此外，在BAL治疗期间，ALF患者血浆中的IgG和IgM抗Galα(1-3)Gal被清除。
猪肝细胞的分离和培养诱导了Galα(1-3)Gal的表达，这可能引发免疫反应，潜在地损害BAL的功能。
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