Characterization of a novel glycolipid with a difucosylated H-antigen in human blood group O erythrocytes with monoclonal antibody HMMC-1 and its detection in human uterine cervical carcinoma tissues
Characterization of a novel glycolipid with a difucosylated H-antigen in human blood group O erythrocytes with monoclonal antibody HMMC-1 and its detection in human uterine cervical carcinoma tissues
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用单克隆抗体 HMMC-1 表征人 O 型红细胞中具有二岩藻糖基化 H 抗原的新型糖脂及其在人宫颈癌组织中的检测
DOI：
10.1007/s10719-019-09873-3
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Iwamori M
中科院分区：
文献类型：
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作者：
Tanaka K;Suzuki A;Aoki D;Iwamori M
用人子宫内膜癌细胞株免疫跨染色体小鼠所制备的人源化单抗HMMC-1，通过共转染O-糖基转移酶和抑制抗体与合成寡糖的结合，与L O-糖链核心上的H抗原发生反应。然而，对不同ABO血型的人红细胞抗鞘糖脂抗体的直接结合分析表明，该抗体能够选择性地与O型血中的极性糖脂结合，而不能与A、B和AB型红细胞中的极性糖脂结合。出乎意料的是，典型的单糖化H-糖脂IV2Fucα-nLC4Cer和VI2Fucα-nLc6Cer是A和B-糖脂的前体，不仅存在于O型血，而且存在于A、B和AB-红细胞中，不是HMMC-1抗体的抗原。该抗原占O型红细胞糖脂总量的0.001%以下，用常规硅胶柱层析纯化。通过全甲基化、GC-MS和ESI-TOFMS的结构测定，证明该糖脂是一种新的糖脂，其末端为Fucα1–2Galβ1-4GlcNAcβ1–3Gal(2-1αFuc)β1-4GlcNAcβ1–3Galβ1-4GlcNAcβ1–3Galβ1-4Glcβ1–1’Cer，-1抗体的表位，其末端为HMMC-1抗体的表位。在29例宫颈癌患者的组织中，有5例检测到HMMC-1糖脂，无论他们的ABO血型如何。
Humanized monoclonal antibody HMMC-1 established by immunizing transchromosomal mice with a human uterine endometrial cancer cell line has been found to react with the H-antigen carried on core l O-glycans through cotransfection of glycosyltransferases for O-glycans and inhibition of antibody-binding with synthetic oligosaccharides. However, direct binding analysis of an antibody against glycosphingolipids from human erythrocytes with different ABO blood groups revealed that it was able to bind selectively with polar glycolipids in blood group O, but not blood group A, B and AB erythrocytes. Unexpectedly, typical monofucosylated H-glycolipids, IV2Fucα-nLc4Cer and VI2Fucα-nLc6Cer, which are the precursors for A and B-glycolipids, and were present not only in blood group O, but also A, B and AB-erythrocytes, were not the antigens for the HMMC-1 antibody. The antigen comprised less than 0.001% of the total glycolipids in blood group O-erythrocytes, and was purified by conventional silica gel column chromatography. Structural determination by permethylation, GC-MS, and ESI-TOFMS demonstrated that the structure was a novel glycolipid with a difucosylated H-antigen, Fucα1–2Galβ1-4GlcNAcβ1–3Gal(2-1αFuc)β1-4GlcNAcβ1–3Galβ1-4GlcNAcβ1–3Galβ1-4Glcβ1–1’Cer, VI2,VIII2(Fucα)2-nLc8Cer, whose terminal difucosylated structure was the epitope of the HMMC-1 antibody. The HMMC-1 glycolipid was detected in five out of 29 tissues from patients suffering from uterine cervical carcinomas, irrespective of their ABO-blood groups.