Reduction of the major xenoantigen on glycosphingolipids of swine endothelial cells by various glycosyltransferases

Masaru Koma, Shuji Miyagawa, Koichi Honke, Yoshitaka Ikeda, Souichi Koyota, Shinichiro Miyoshi, Hikaru Matsuda, Shuichi Tsuji, Ryota Shirakura, Naoyuki Taniguchi

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24 Citations (Scopus)

Abstract

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 IB4 (GSIB4) lectin, which binds to the Gal α1-3 Galβ 1-4 GlcNAc-R (α-galactosyl epitope), was reduced by ~20% by the treatment with D-PDMP (D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol), suggesting that glycosphingolipids contained by SEC have a considerable amount of the α-galactosyl epitope. The overexpression of two different types of glycosyltransferase, N-acetylglucosaminyl transferase III (GnT-III), as well as α2,6-sialyltransferase (ST6Gal I), α2,3-sialyltransferase (ST3Gal III), and α1,2-fucosyltransferase (α1,2FT), 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 α-galactosyl epitope in glycosphingolipids were not decreased in the GnT-III transfectants but were in the ST6Gal I, ST3Gal III, and α1,2FT transfectants. These data indicate that ST6Gal I, ST3Gal III, and α1,2FT reduced the α-galactosyl epitope in both glycoproteins and glycosphingolipids, while GnT-III reduced them only in glycoproteins.

Original languageEnglish
Pages (from-to)745-751
Number of pages7
JournalGlycobiology
Volume10
Issue number7
DOIs
Publication statusPublished - Jan 1 2000

Keywords

  • Glycosphingolipids
  • Glycosyltransferase
  • Swine endothelial cell
  • The α-galactosyl epitope
  • Xenotransplantation

ASJC Scopus subject areas

  • Biochemistry

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    Koma, M., Miyagawa, S., Honke, K., Ikeda, Y., Koyota, S., Miyoshi, S., Matsuda, H., Tsuji, S., Shirakura, R., & Taniguchi, N. (2000). Reduction of the major xenoantigen on glycosphingolipids of swine endothelial cells by various glycosyltransferases. Glycobiology, 10(7), 745-751. https://doi.org/10.1093/glycob/10.7.745