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

Kyoko Tanaka, Akemi Suzuki, Daisuke Aoki, Masao Iwamori

Research output: Contribution to journalArticle

Abstract

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, IV 2 Fucα-nLc 4 Cer and VI 2 Fucα-nLc 6 Cer, 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, VI 2 ,VIII 2 (Fucα) 2 -nLc 8 Cer, 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.

Original languageEnglish
JournalGlycoconjugate Journal
DOIs
Publication statusPublished - 2019 Jan 1

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Glycolipids
Blood Group Antigens
Erythrocytes
Tissue
Carcinoma
Antibodies
Polysaccharides
Antigens
Glycosphingolipids
Glycosyltransferases
Uterine Neoplasms
Column chromatography
Silica Gel
Endometrial Neoplasms
Oligosaccharides
Gel Chromatography
HMMC-1 monoclonal antibody
H antigen
Epitopes
Cells

Keywords

  • Antigenic determinant
  • Carbohydrate structure
  • Glycosphingolipids
  • Monoclonal antibody HMMC-1
  • Uterine cervical carcinoma

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

@article{717ba62d99d24a9fb5a3493e860e8fde,
title = "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",
abstract = "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, IV 2 Fucα-nLc 4 Cer and VI 2 Fucα-nLc 6 Cer, 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, VI 2 ,VIII 2 (Fucα) 2 -nLc 8 Cer, 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.",
keywords = "Antigenic determinant, Carbohydrate structure, Glycosphingolipids, Monoclonal antibody HMMC-1, Uterine cervical carcinoma",
author = "Kyoko Tanaka and Akemi Suzuki and Daisuke Aoki and Masao Iwamori",
year = "2019",
month = "1",
day = "1",
doi = "10.1007/s10719-019-09873-3",
language = "English",
journal = "Glycoconjugate Journal",
issn = "0282-0080",
publisher = "Springer Netherlands",

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TY - JOUR

T1 - 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

AU - Tanaka, Kyoko

AU - Suzuki, Akemi

AU - Aoki, Daisuke

AU - Iwamori, Masao

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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, IV 2 Fucα-nLc 4 Cer and VI 2 Fucα-nLc 6 Cer, 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, VI 2 ,VIII 2 (Fucα) 2 -nLc 8 Cer, 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.

AB - 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, IV 2 Fucα-nLc 4 Cer and VI 2 Fucα-nLc 6 Cer, 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, VI 2 ,VIII 2 (Fucα) 2 -nLc 8 Cer, 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.

KW - Antigenic determinant

KW - Carbohydrate structure

KW - Glycosphingolipids

KW - Monoclonal antibody HMMC-1

KW - Uterine cervical carcinoma

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