Reduction of the major xenoantigen on glycosphingolipids of swine endothelial cells by various glycosyltransferases.-猫眼课题宝

通过各种糖基转移酶减少猪内皮细胞鞘糖脂上的主要异种抗原。

DOI：

发表时间：

2000

期刊：

Glycobiology

影响因子：

4.3

通讯作者：

中科院分区：

生物学3区

文献类型：

作者：

M. Koma;Shuji Miyagawa;Koichi Honke;Yoshitaka Ikeda;S. Koyota;S. Miyoshi;Hikaru Matsuda;Shuichi Tsuji;R. Shirakura;Naoyuki Taniguchi

使用转染的猪内皮细胞（SEC）系检查各种糖基转移酶对鞘糖脂的影响。亲本SEC对正常人血清（NHS）和与Gal α 1 - 3 Gal β 1 - 4 GlcNAc-R结合的Griffonia simplicifolia IB（4）（GSIB 4）凝集素的反应性（α-半乳糖基表位），通过用D-PdR处理减少了约20（D-苏型-1-苯基-2-癸酰基氨基-3-吗啉代-1-丙醇），表明SEC所含的鞘糖脂具有相当大量的α-半乳糖基表位。两种不同类型的糖基转移酶，N-乙酰葡糖胺基转移酶III（GnT-III），以及α 2，6-唾液酸转移酶（ST6Gal I）、α 2，3-唾液酸转移酶（ST3Gal III）和α 1，2-岩藻糖基转移酶（α 1，2FT）的过表达显著抑制SEC的总抗原性。然而，在GnT-III转染子的情况下，对NHS和GSIB4凝集素的反应性的减少比其他转染子中的那些更温和。Western印迹分析表明，所有转染子中的糖蛋白对NHS和GSIB 4凝集素的反应性降低到大致相同的程度。因此，通过薄层色谱法分离这些转染子的中性鞘糖脂，然后用NHS和GSIB 4凝集素进行免疫染色。在GnT-III转染子中，鞘糖脂中α-半乳糖基表位的水平没有降低，但在ST6Gal I、ST3Gal III和α 1，2FT转染子中降低。这些数据表明，ST6Gal I、ST3Gal III和α 1，2FT减少糖蛋白和鞘糖脂中的α-半乳糖基表位，而GnT-III仅减少糖蛋白中的α-半乳糖基表位。

The effect of the various glycosyltransferases on glycosphingolipids was examined, using transfected swine endothelial cell (SEC) lines. The reactivity of parental SEC to normal human serum (NHS) and Griffonia simplicifolia IB(4) (GSIB4) lectin, which binds to the Gal alpha1-3 Gal beta 1-4 GlcNAc-R (alpha-galactosyl epitope), was reduced by approximately 20% by the treatment with D-PDMP (D-threo-1-phenyl-2-decan- oylamino-3-morpholino-1-propanol), suggesting that glycosphingolipids contained by SEC have a considerable amount of the alpha-galactosyl epitope. The overexpression of two different types of glycosyltransferase, N-acetylglucosaminyl transferase III (GnT-III), as well as alpha2, 6-sialyltransferase (ST6Gal I), alpha2,3-sialyltransferase (ST3Gal III), and alpha1,2-fucosyltransferase (alpha1,2FT), suppresses the total antigenicity of SEC significantly. However, the reduction in reactivities toward NHS and GSIB4 lectin in the case of GnT-III transfectants was milder than those in other transfectants. Western blot analysis indicated that the glycoproteins in all transfectants had diminished reactivity to NHS and GSIB4 lectin to approximately the same extent. Therefore, the neutral glycosphingolipids of these transfectants were separated by thin layer chromatography, followed by immunostaining with NHS and GSIB4 lectin. The levels of the alpha-galactosyl epitope in glycosphingolipids were not decreased in the GnT-III transfectants but were in the ST6Gal I, ST3Gal III, and alpha1,2FT transfectants. These data indicate that ST6Gal I, ST3Gal III, and alpha1,2FT reduced the alpha-galactosyl epitope in both glycoproteins and glycosphingolipids, while GnT-III reduced them only in glycoproteins.