This work was supported by People's Hospital of Zhejiang Province.
猪心移植是解决心脏移植供体少的有效方法.由于猪心血管内皮细胞表面抗原半乳糖-α-1,3-半乳糖(Gal-α-1,3-Gal),能与人体内预存的天然抗体结合产生超急性排斥反应(HAR),而无法应用于临床.为了解决这一难题, 应用噬菌体展示技术,筛选出一个能与抗B型血单克隆抗体(mAB anti-B) 特异性结合的阳性噬菌体克隆,应用酶联免疫吸附测定(ELISA)和竞争性ELISA结果表明,所获得的阳性噬菌体克隆能特异性地与mAB anti-B结合,并且这种结合可被蜜二糖(具有Gal-α-1,6-Glc的结构)所竞争抑制.由此推测,筛选到的噬菌体阳性克隆很可能就结合在蜜二糖与mAB anti-B结合的位点.同时,进行了阳性噬菌体克隆的抑制猪红细胞凝集活性试验,试验结果表明,此阳性克隆不仅可抑制Gal-α-1,3-Gal与抗体的结合, 也可抑制Gal-α-1,3-Gal与西非单叶豆凝集素(GS-I-B4)的结合.此噬菌体阳性克隆测序后,得小肽序列为CCWLLRQPVRFVRSIRS.鉴于以上的结果,认为此小肽可以作为Gal-α-1,3-Gal的模拟肽,同时有望开发成抗猪器官异种移植超急性排斥反应的新药.
The obstacle for pig-to-human transplantation is hyperacute rejection (HAR) triggered by the interaction between human natural antibodies and the antigenic epitope Gal-α-1,3-Gal. The α-Gal structure is considered to be the major xenoantigenic epitopes present on porcine tissues. A phage-displayed peptide library is used to identify a 17-amino-acid peptide CCWLLRQPVRFVRSIRS that binds to the mAb anti-B (anti-B monoclonal antibody which binds the carbohydrate Gal α-1,3 Gal).The melibiose competes with the binding of mAB anti-B to the peptide, suggesting that they may bind the same site. Using a pig RBC agglutination assay, it is shown that this peptide can inhibit the agglutination of pig RBCs by human serum or GS-I-B4.
张晖,詹金彪,许林海,严志焜,王克夷.用噬菌体展示筛选Gal-α-1,3-Gal的模拟肽[J].生物化学与生物物理进展,2003,30(3):461-465
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