免疫组织化学法特异性检测动物组织中的 Neu5Gc。

Specific Detection of Neu5Gc in Animal Tissues by Immunohistochemistry.

机构信息

Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, 69978, Israel.

出版信息

Methods Mol Biol. 2020;2110:59-72. doi: 10.1007/978-1-0716-0255-3_4.

DOI:10.1007/978-1-0716-0255-3_4

PMID:32002901

Abstract

One of the major obstacles in xenotransplantation is tissue expression of the non-human mammalian carbohydrate N-glycolylneuraminic acid (Neu5Gc). This 9-carbon backbone acidic sugar is the hydroxylated form of N-acetylneuraminic acid (Neu5Ac), and both constitute the two most common sialic acid types in mammals. Loss of CMP-Neu5Ac hydroxylase encoding gene in humans dictates the immunogenic nature of Neu5Gc-containing xenografts. Here we describe an immunohistochemistry method for the detection of Neu5Gc in mammalian-derived tissues using affinity-purified chicken anti-Neu5Gc IgY. Specificity is further demonstrated by competitive inhibition with free Neu5Gc or Neu5Gc-glycoproteins, but not with Neu5Ac or Neu5Ac-glycoproteins. This method can be used to evaluate potential Neu5Gc-immunogenicity of xenografts.

摘要

异种移植的主要障碍之一是组织表达非人类哺乳动物碳水化合物 N-羟乙酰神经氨酸 (Neu5Gc)。这种 9 碳骨架酸性糖是 N-乙酰神经氨酸 (Neu5Ac) 的羟化形式，两者都是哺乳动物中最常见的两种唾液酸类型。人类 CMP-Neu5Ac 羟化酶编码基因的缺失决定了含有 Neu5Gc 的异种移植物的免疫原性。在这里，我们描述了一种使用亲和纯化的鸡抗 Neu5Gc IgY 检测哺乳动物来源组织中 Neu5Gc 的免疫组织化学方法。通过与游离 Neu5Gc 或 Neu5Gc-糖蛋白的竞争性抑制进一步证明了特异性，但不能与 Neu5Ac 或 Neu5Ac-糖蛋白。该方法可用于评估异种移植物潜在的 Neu5Gc 免疫原性。