Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1984 May 1;219(3):865–874. doi: 10.1042/bj2190865

The reactivities of human erythrocyte autoantibodies anti-Pr2, anti-Gd, Fl and Sa with gangliosides in a chromatogram binding assay.

K Uemura, D Roelcke, Y Nagai, T Feizi
PMCID: PMC1153555  PMID: 6204642

Abstract

The thin layer chromatogram binding assay was used to study the reaction of several natural-monoclonal autoantibodies which recognize sialic acid-dependent antigens of human erythrocytes. Immunostaining of gangliosides derived from human and bovine erythrocytes was achieved with four autoantibodies designated anti-Pr2, anti-Gd, Sa and Fl, each of which has a different haemagglutination pattern with untreated and proteinase-treated erythrocytes and with cells of I and i antigen types. From the chromatogram binding patterns of anti-Pr2 with gangliosides of the neolacto and the ganglio series, it is deduced that this antibody reacts best with N-acetylneuraminic acid when it is alpha 2-3- or alpha 2-6-linked to a terminal Gal(beta 1-4)Glc/GlcNAc GlcNAc sequence and to a lesser extent when it is alpha 2-3-linked to a terminal Gal(beta 1-3)GalNAc sequence or to an internal galactose and when it is alpha 2-8-linked to another, internal N-acetylneuraminic acid residue. The other three antibodies differ from anti-Pr2 in their lack of reaction with glycolipids of the ganglio series. They react with the NeuAc(alpha 2-3)Gal(beta 1-4)Glc/GlcNAc sequence as found in GM3 and in glycolipids of the neolacto series, but show a preference for the latter, longer sequences. Thus all four antibodies react with sialylated oligosaccharides containing i type (linear) and I type (branched) neolacto backbones. Fl antibody differs from the other three in its stronger reaction with branched neolacto sequences in accordance with its stronger agglutination of erythrocytes of I rather than i type. The four antibodies show a specificity for N-acetyl- rather than N-glycolyl-neuraminic acid.

Full text

PDF
865

Images in this article

868Fig. 1.
on p.868
869Fig. 2.
on p.869
869Fig. 3.
on p.869
870Fig. 4.
on p.870

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chien J. L., Li S. C., Laine R. A., Li Y. T. Characterization of gangliosides from bovine erythrocyte membranes. J Biol Chem. 1978 Jun 10;253(11):4031–4035. [PubMed] [Google Scholar]
  2. DODGE J. T., MITCHELL C., HANAHAN D. J. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963 Jan;100:119–130. doi: 10.1016/0003-9861(63)90042-0. [DOI] [PubMed] [Google Scholar]
  3. Ebert W., Fey J., Gärtner C., Geisen H. P., Rautenberg U., Roelcke D., Weicker H. Isolation and partial characterization of the Pr autoantigen determinants. Mol Immunol. 1979 Jun;16(6):413–419. doi: 10.1016/0161-5890(79)90109-3. [DOI] [PubMed] [Google Scholar]
  4. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  5. Feizi T., Childs R. A., Hakomori S. I., Powell M. E. Blood-group-Ii-active gangliosides of human erythrocyte membranes. Biochem J. 1978 Jul 1;173(1):245–254. doi: 10.1042/bj1730245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feizi T., Childs R. A., Watanabe K., Hakomori S. I. Three types of blood group I specificity among monoclonal anti-I autoantibodies revealed by analogues of a branched erythrocyte glycolipid. J Exp Med. 1979 Apr 1;149(4):975–980. doi: 10.1084/jem.149.4.975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Feizi T. The antigens Ii, SSEA-1 and ABH are in interrelated system of carbohydrate differentiation antigens expressed on glycosphingolipids and glycoproteins. Adv Exp Med Biol. 1982;152:167–177. [PubMed] [Google Scholar]
  8. Feizi T. The blood group Ii system: a carbohydrate antigen system defined by naturally monoclonal or oligoclonal autoantibodies of man. Immunol Commun. 1981;10(2):127–156. doi: 10.3109/08820138109050693. [DOI] [PubMed] [Google Scholar]
  9. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hakomori S. Glycosphingolipids in cellular interaction, differentiation, and oncogenesis. Annu Rev Biochem. 1981;50:733–764. doi: 10.1146/annurev.bi.50.070181.003505. [DOI] [PubMed] [Google Scholar]
  11. Kannagi R., Roelcke D., Peterson K. A., Okada Y., Levery S. B., Hakomori S. Characterization of an epitope (determinant) structure in a developmentally regulated glycolipid antigen defined by a cold agglutinin Fl, recognition of alpha-sialosyl and alpha-L-fucosyl groups in a branched structure. Carbohydr Res. 1983 Aug 16;120:143–157. doi: 10.1016/0008-6215(83)88013-6. [DOI] [PubMed] [Google Scholar]
  12. Kawamura N., Taketomi T. A new procedure for the isolation of brain gangliosides, and determination of their long chain base compositions. J Biochem. 1977 May;81(5):1217–1225. [PubMed] [Google Scholar]
  13. Kundu S. K., Marcus D. M., Roelcke D. Glycosphingolipid receptors for anti-Gd and anti-p cold agglutinins. Immunol Lett. 1982 May;4(5):263–267. doi: 10.1016/0165-2478(82)90049-9. [DOI] [PubMed] [Google Scholar]
  14. Kunishita T., Uemura K., Okano A., Taketomi T. Isolation of basic protein-acidic lipid complex from myelin. Jpn J Exp Med. 1979 Dec;49(6):391–396. [PubMed] [Google Scholar]
  15. Ledeen R. W., Yu R. K., Eng L. F. Gangliosides of human myelin: sialosylgalactosylceramide (G7) as a major component. J Neurochem. 1973 Oct;21(4):829–839. doi: 10.1111/j.1471-4159.1973.tb07527.x. [DOI] [PubMed] [Google Scholar]
  16. Macher B. A., Lee W. M., Westrick M. A. Glycosphingolipids of normal and leukemic human leukocytes. Mol Cell Biochem. 1982 Sep 3;47(2):81–95. doi: 10.1007/BF00234409. [DOI] [PubMed] [Google Scholar]
  17. Magnani J. L., Brockhaus M., Smith D. F., Ginsburg V., Blaszczyk M., Mitchell K. F., Steplewski Z., Koprowski H. A monosialoganglioside is a monoclonal antibody-defined antigen of colon carcinoma. Science. 1981 Apr 3;212(4490):55–56. doi: 10.1126/science.7209516. [DOI] [PubMed] [Google Scholar]
  18. Momoi T., Ando S., Magai Y. High resolution preparative column chromatographic system for gangliosides using DEAE-Sephadex and a new porus silica, Iatrobeads. Biochim Biophys Acta. 1976 Sep 27;441(3):488–497. [PubMed] [Google Scholar]
  19. Niemann H., Watanabe K., Hakomori S. Blood group i and I activities of "lacto-N-norhexaosylceramide" and its analogues: the structural requirements for i-specificities. Biochem Biophys Res Commun. 1978 Apr 28;81(4):1286–1293. doi: 10.1016/0006-291x(78)91275-5. [DOI] [PubMed] [Google Scholar]
  20. Roelcke D. A further cold agglutinin, F1, recognizing a N-acetylneuraminic acid-determined antigen. Vox Sang. 1981;41(2):98–101. doi: 10.1111/j.1423-0410.1981.tb01021.x. [DOI] [PubMed] [Google Scholar]
  21. Roelcke D., Brossmer R., Riesen W. Inhibition of human anti-Gd cold agglutinins by sialyllactose. Scand J Immunol. 1978;8(3):179–185. doi: 10.1111/j.1365-3083.1978.tb00508.x. [DOI] [PubMed] [Google Scholar]
  22. Roelcke D. Cold agglutination. Antibodies and antigens. Clin Immunol Immunopathol. 1974 Jan;2(2):266–280. doi: 10.1016/0090-1229(74)90044-0. [DOI] [PubMed] [Google Scholar]
  23. Roelcke D., Ebert W., Geisen H. P. Anti-Pr3: serological and immunochemical identification of a new anti-Pr subspecificity. Vox Sang. 1976;30(2):122–133. doi: 10.1111/j.1423-0410.1976.tb02802.x. [DOI] [PubMed] [Google Scholar]
  24. Roelcke D., Pruzanski W., Ebert W., Römer W., Fischer E., Lenhard V., Rauterberg E. A new human monoclonal cold agglutinin Sa recognizing terminal N-acetylneuraminyl groups on the cell surface. Blood. 1980 Apr;55(4):677–681. [PubMed] [Google Scholar]
  25. Roelcke D., Riesen W., Geisen H. P., Ebert W. Serological identification of the new cold agglutinin specificity anti-Gd. Vox Sang. 1977;33(5):304–306. doi: 10.1111/j.1423-0410.1977.tb04480.x. [DOI] [PubMed] [Google Scholar]
  26. Roelcke D. Serological studies on the Pr 1 -Pr 2 antigens using dog erythrocytes. Differentiation of Pr 2 from Pr 1 and detection of a Pr 1 heterogeneity: Pr 1h -Pr 1d . Vox Sang. 1973 Apr;24(4):354–361. doi: 10.1111/j.1423-0410.1973.tb02653.x. [DOI] [PubMed] [Google Scholar]
  27. Roelcke D. Specificity of IgA cold agglutinins: anti-Pr 1 . Eur J Immunol. 1973 Apr;3(4):206–212. doi: 10.1002/eji.1830030406. [DOI] [PubMed] [Google Scholar]
  28. Roelcke D. The Lud cold agglutinin: a further antibody recognizing N-acetylneuraminic acid-determined antigens not fully expressed at birth. Vox Sang. 1981 Nov-Dec;41(5-6):316–318. doi: 10.1111/j.1423-0410.1981.tb01056.x. [DOI] [PubMed] [Google Scholar]
  29. Römer W., Seelig H. P., Lenhard V., Roelcke D. The distribution of I/i, Pr and Gd antigens in mammalian tissues. Invest Cell Pathol. 1979 Jul-Sep;2(3):157–162. [PubMed] [Google Scholar]
  30. SVENNERHOLM L. CHROMATOGRAPHIC SEPARATION OF HUMAN BRAIN GANGLIOSIDES. J Neurochem. 1963 Sep;10:613–623. doi: 10.1111/j.1471-4159.1963.tb08933.x. [DOI] [PubMed] [Google Scholar]
  31. Siddiqui B., Hakomori S. A ceramide tetrasaccharide of human erythrocyte membrane reacting with anti-type XIV pneumococcal polysaccharide antiserum. Biochim Biophys Acta. 1973 Dec 13;330(2):147–155. doi: 10.1016/0005-2736(73)90219-8. [DOI] [PubMed] [Google Scholar]
  32. Uemura K., Childs R. A., Hanfland P., Feizi T. A multiplicity of erythrocyte glycolipids of the neolacto series revealed by immuno-thin-layer chromatography with monoclonal anti-I and anti-i antibodies. Biosci Rep. 1983 Jun;3(6):577–588. doi: 10.1007/BF01120703. [DOI] [PubMed] [Google Scholar]
  33. Uemura K., Yuzawa M., Taketomi T. Characterization of major glycolipids in bovine erythrocyte membrane. J Biochem. 1978 Feb;83(2):463–471. doi: 10.1093/oxfordjournals.jbchem.a131933. [DOI] [PubMed] [Google Scholar]
  34. Watanabe K., Hakomori S. I., Childs R. A., Feizi T. Characterization of a blood group I-active ganglioside. Structural requirements for I and i specificities. J Biol Chem. 1979 May 10;254(9):3221–3228. [PubMed] [Google Scholar]
  35. Watanabe K., Powell M. E., Hakomori S. I. Isolation and characterization of gangliosides with a new sialosyl linkage and core structures. II. Gangliosides of human erythrocyte membranes. J Biol Chem. 1979 Sep 10;254(17):8223–8229. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES