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Removal of Alpha-Gal Epitopes from Porcine Aortic Valve and Pericardium using Recombinant Human Alpha Galactosidase A

机译:使用重组人α半乳糖苷酶A去除猪主动脉瓣和心包中的Al-Gal表位

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摘要

It has been reported that the immune response due to α-Gal epitopes is an important factor in tissue valve failure. The elimination of the interaction between the natural anti-Gal antibodies and α-gal epitopes on the xenografts is a prerequisite to the success of xenografts in humans. Previously, we reported that the green coffee bean α-galactosidase could remove all α-Gal epitopes from cell surface of porcine aortic valve and pericardial tissue, but it has limitations on cost effectiveness. In this study we wanted to know whether the recently produced recombinant human α-galactosidase A has the same effective enzymatic activity as green coffee bean α-galactosidase in removing α-Gal epitopes from the same tissues. After treating fresh porcine aortic valve and pericardial tissue with recombinant α-galactosidase A, each sample was stained with Griffonia simplicifolia type I isolectin B4 indirect immunoperoxidase avidin-biotin technique. We then examined whether the α-Gal epitopes were reduced or abolished in each consecutive concentration of recombinant α-galactosidase A by comparing the degree of the Griffonia simplicifolia isolectin B4 staining. As a result, the recombinant α-galactosidase A could remove cell surface α-Gals on porcine aortic valve and pericardial tissue as effectively as green coffee bean α-galactosidase.
机译:据报道,由于α-Gal表位引起的免疫应答是组织瓣膜衰竭的重要因素。消除天然抗-Gal抗体与异种移植物上的α-gal表位之间的相互作用是人类异种移植物成功的先决条件。以前,我们曾报道绿色咖啡豆α-半乳糖苷酶可以去除猪主动脉瓣和心包组织细胞表面的所有α-Gal表位,但其成本效益受到限制。在这项研究中,我们想知道最近生产的重组人α-半乳糖苷酶A在去除相同组织中的α-Gal表位方面是否具有与生咖啡豆α-半乳糖苷酶相同的有效酶活性。用重组α-半乳糖苷酶A处理新鲜的猪主动脉瓣和心包组织后,每个样品均用Griffonia simplicifolia I型异凝集素B4间接免疫过氧化物酶亲和素-生物素技术染色。然后,我们通过比较辛普利叶单胞菌isolectin B4染色的程度,检查了重组α-半乳糖苷酶A的每个连续浓度中α-Gal表位是否被减少或消除。结果,重组α-半乳糖苷酶A可以像生咖啡豆α-半乳糖苷酶一样有效地去除猪主动脉瓣和心包组织上的细胞表面α-Gal。

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