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为了测定猪血管内皮细胞膜表面可能存在的碳水化合物抗原表位,5个人工合成的糖结合物(还原胺化法)和猪胃粘膜糖肽片断被用于中和人天然抗体.然后再用这类中和之后的人血清与猪血管内皮细胞行补体依赖的细胞毒反应(MTT法),并与牛血清白蛋白对比.有2个糖结合物能部分和人天然素体,它们是:(1)去唾液酸分枝双糖-牛血清白蛋白;(2)Le~a-Le~x钥孔虫戚血蓝素.这些糖结合物均含有Gal(β1,3)GlcNAc或Gal(β1,4)GlcNAc.猪胃粘膜糖肽片断具有较强地抑制人血清细胞毒性的作用,除含有Gal(β1,3)GlcNAc、Gal(β1,4)GlcNAc之外,它的主要结构为GlcNAc(χ1,4)Gal.用钥孔虫戚血蓝素作载体能加强糖结合物中和人天然抗体的作用.提示天然抗体认别成簇状的糖表位结构.
In order to determine possible carbohydrate epitopes on the surface of porcine vascular endothelial cells, five synthetic sugar conjugates (reductive amination) and porcine gastric mucosa glycopeptide fragments were used to neutralize human natural antibodies, Complementary and human serum and porcine vascular endothelial cells undergo a complement-dependent cytotoxic reaction (MTT assay) and are comparable to bovine serum albumin (BSA). Two glycoconjugates are available to partially and naturally-occurring human bodies: 1) Asialo-branched disaccharide - bovine serum albumin; (2) Le ~ a-Le ~ x keyhole limpet hemocyanin These carbohydrate conjugates all contain Gal (β1,3) GlcNAc or Gal , 4) GlcNAc. The porcine gastric mucosa glycopeptide fragment has a strong inhibitory effect on human serum cytotoxicity. Its main structure except Gal (β1,3) GlcNAc, Gal (β1,4) GlcNAc is GlcNAc χ1, 4) Gal. The use of keyhole limpet hemocyanin as a carrier enhances the neutralizing effect of the glycoconjugate on human natural antibodies, suggesting that natural antibodies recognize clustered sugar epitope structures.