Summary
Endo-β-galactosidase from Escherichia freundii cleaves polylactosaminyl structures as follows: R-GlcNAcβ1-3Galβ1-4GlcNAcβ1-R′ + H2O → R-GlcNAcβ1–3Gal + GlcNAcβ1-R′. By staining with Griffonia simplicifolia agglutinin-II following the enzyme digestion, the distribution of R-GlcNAcβ1–3Galβ1–4GlcNAc can be demonstrated in tissue sections. This carbohydrate chain is one of the backbone structures carrying the blood-group-related antigens and, thus, localization of this structure may provide detailed information about the distribution of variants with different backbone structures. Various formalin-fixed, paraffin-embedded tissue sections were stained by Griffonia simplicifolia agglutinin-II with or without prior enzyme digestion and the reactivity of the agglutinin imparted by enzyme digestion was studied in the following tissues and cells: pancreatic acinar cells, gastric surface mucosae, duct cells and mucous cells of salivary glands and tracheal glands, surface epithelium of trachea, goblet cells of large intestine, columnar epithelium of uterine cervical glands, distal and collecting tubules of kidney, certain cells of anterior lobe and colloid of middle lobe of pituitary glands, epithelial reticular cells and Hassall's corpuscles of thymus and Kupffer cells of liver. In gastric surface mucosae, the reactivity of the agglutinin appeared in non-secretor individuals but not in the secretor individuals, and in mucous cells of salivary and tracheal glands the reactivity appeared in Le(a - b -) non-secretor individuals but not in Le(a + b -) non-secretor or secretor individuals. In pancreatic acinar cells and duct cells of salivary glands from fetuses and newborn infants, prior fucosidase digestion markedly enhanced the Griffonia simplicifolia agglutinin-II reactivity elicited by endo-β-galactosidase digestion. Prior fucosidase digestion was also a prerequisite for revealing the reactivity of this agglutinin by endo-β-galactosidase digestion in gastric surface mucosae from secretor individuals. β-Galactosidase digestion disclosed reactivity of this agglutinin in pancreatic acinar cells and duct cells of salivary glands even after the removal of endo-β-galactosidase-labile lactosamine structures by sequential digestion with endo-β-galactosidase and β-N-acetylhexosaminidase. These results demonstrate that the procedures developed in this study provide a useful means for detecting different types of lactosamine structures which carry blood-group antigens in humans tissues.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Carlsson, S. R. & Fukuda, M. (1990) The polylactosaminoglycans of human lysosomal membrane glycoproteins lamp-1 and lamp-2. J. Biol Chem. 265, 20 488–95.
Chen, H. T. & Kabat, E. A. (1985) Imunochemical studies on blood groups. The combining site specificities of mouse monoclonal hybridoma anti-A and anti-B. J. Biol. Chem. 260, 13 208–17.
Clausen, H. & Hakomori, S.-H. (1989) ABH and related histo-blood group antigens; immunochemical differences in carrier isotypes and distribution. Vox Sang. 56, 1–20.
Clausen, H., Levery, S. B., Dabelsteen, E. & Hakomori, S. (1987) Blood group ABH antigens: a new series of blood group A-associated structures (genetic regulation and tissue distribution). Transplant. Proc. 19, 4408–12.
Ebisu, S., Iyer, S. & Goldstein, I. J. (1978) Equilibrium dialysis and carbohydrate-binding studies on the 2-acetimido-2- deoxy-d-glucopyranosyl-binding lectin from Bandeiraea simplicifolia seeds. Carbohydr. Res. 61, 129–38.
Feizi, T. (1981) The blood group Ii System: a carbohydrate antigen system defined by natural monoclonal or oligoclonal autoantibodies of man. Immunolog. Commun. 10, 127–56.
Feizi, T. (1985) Demonstration by monoclonal antibodies that carbohydrate structures of glycoproteins and glycolipids are onco-developmental antigens. Nature 314, 53–7.
Fukuda, M. (1985) Cell surface glycoconjugates as oncodifrerentiation markers in hematopoietic cells. Biochim. Biophys. Acta 780, 119–50.
Fukuda, M. N. & Matsumura, G. (1976) Endo-β-galactosidase of Escherichia freundii. Purification and endoglycosidic action on keratan sulfates, oligosaccharides, and blood group active glycoprotein. J. Biol. Chem. 251, 6218–25.
Fukuda, M. N., Watanabe, K. & Hakomori, S. (1978) Release of oligosaccharides from various glycosphingolipids by endo-β-galactosidase. J. Biol. Chem. 253, 814–19.
Fukuda, M., Dell, A., Oates, J. E. & Fukuda, M. N. (1984a) Structure of branched lactosaminoglycan, the carbohydrate moiety of band 3 isolated from adult human erythrocytes. J. Biol. Chem. 259, 8260–73.
Fukuda, M., Spooncer, E., Oates, J. E., Dell, A. & Klock, J. C. (1984b) Structure of sialylated fucosyl lactosaminoglycan isolated from human granulocytes. J. Biol. Chem. 259, 10 925–35.
Fukuda, M., Carlsson, S. R., Klock, J. C. & Dell, A. (1986) Structures of O -linked oligosaccharides isolated from normal granulocytes, chronic myelogenous leukemia cells, and acute myelogenous leukemia cells. J. Biol. Chem. 261, 12 796–806.
Gane, P., Vallayoudom, J., Mollicone, R., Breimer, M. E., Samuelsson, B. E., Rouger, P., Gerard, G., LePendu, J. & Oriol, R. (1987) Heterogeneity of anti-A and anti-B monoclonal reagents. Agglutination of some weak ABH erythrocyte variants and recognition of synthetic oligosaccharide and tissue antigens. Vox Sang. 53, 117–25.
Griffin, N. R. & Wells, M. (1993) Semiquantitative immunohistochemical studies of blood group antigens A, B, H, Lea, Leb, structures and Ii backbone chains in the normal human cervix and in cervical adenocarcinoma. Histochem. J. 25, 228–41.
Hakomori, S. (1989) Aberrant glycosylation in tumors and tumor-associated carbohydrate antigens. Adv. Cancer Res. 52, 257–331.
Hirohashi, S., Ino, Y., Kodama, T. & Shimosato, Y. (1984) Distribution of blood group antigens A, B, H, and I (Ma) in mucous-producing adenocarcinoma of human lung. J. Natl Canc. Inst. 72, 1299–305.
Hubbard, S. C. (1987) Differential effects of oncogenic transformation on N-linked oligosaccharide processing at individual glycosylation sites of viral glycoproteins. J. Biol. Chem. 262, 16 403–11.
Ito, N. & Hirota, T. (1992) Histochemical and cytochemical localization of blood group antigens. Progr. Histochem. Cytochem. 25, 1–85.
Ito, N., Nishi, K., Nakajima, M., Matsuda, Y., Ishitani, A., Mizumoto, J. & Hirota, T. (1986) Localization of blood group antigens in human pancreas with lectin-horseradish peroxidase conjugates. Acta Histochem. Cytochem. 19, 205–18.
Ito, N., Nishi, K., Nakajima, M., Ishitani, A., Okamura, Y., Matsuda, Y. & Hirota, T. (1987) Histochemical reactivity of soybean agglutinin with blood group antigens and their precursor substances in acinar cells of human pancreas. J. Histochem. Cytochem. 35, 881–90.
Ito, N., Nishi, K., Nakajima, M., Okamura, Y. & Hirota, T. (1988) Effects of α-l-fucosidase digestion on lectin staining in human pancreas. J. Histochem. Cytochem. 36, 503–9.
Ito, N., Nishi, K., Nakajima, M., Okamura, Y. & Hirota, T. (1989a) Histochemical analysis of the chemical structure of blood group-related carbohydrate chains in serous cells of human submandibular glands using lectin staining and glycosidase digestion. J. Histochem. Cytochem. 37, 1115–24.
Ito, N., Nishi, K., Nakajima, M., Okamura, Y. & Hirota, T. (1989b) Histochemical demonstration of O-glycosidically linked, type 3 based ABH antigens in human pancreas using lectin staining and glycosidase digestion procedures. Histochemistry 92, 307–12.
Kannagi, R., Nudelman, E., Levery, S. B. & Hakomori, S. (1982) A series of human erythrocyte glycosphingolipids reacting to the monoclonal antibody directed to a developmentally regulated antigen, SSEA-1. J. Biol. Chem. 257, 14 865–74.
Kapadia, A., Feizi, T., Jewell, D., Keeling, J. & Sloven, G. (1981) Immunocytochemical studies of blood group A, H, I, and i antigens in gastric mucosae of infants with normal gastric histology and of patients with gastric carcinoma and chronic benign peptic ulceration. J. Clin. Pathol. 34, 320–37.
Lee, N., Wang, W.-C. & Fukuda, M. (1990) Granulocytic differentiation of HL-60 cells is associated with increase of poly-N-acetyllactosamine in Asn-linked oligosaccharides attached to human lysosomal membrane glycoproteins. J. Biol. Chem. 265, 20 476–87.
Maley, F., Trimble, R. B., Tarentino, A. L. & Plummer, Jr. T. H. (1989) Characterization of glycoproteins and their associated oligosaccharides through the use of endoglycosidases. Anal. Biochem. 180, 195–204.
Murata, H., Takahashi, N. & Yamada, K. (I984) Effects of digestion with N-oligosaccharide glucopeptidase upon certain lectin-peroxidase-diaminobenzidine reactions of glycoproteins in mammalian and avain tissues. Histochemistry 81, 401–7.
Oriol, R. (1993) Molecular basis of major blood group antigens. In Blood Cell Biochemistry, vol. 6 (edited by Cartron, J. P. & Rouger, P.) New York: Plenum Press (in press).
Oriol, R., LePendu, J. & Mollicone, R. (1986) Genetics of ABO, H. Lewis, X and related antigens. Vox Sang. 51, 161–71.
Oriol, R., Samuelsson, B. E. & Messeter, L. (1990) ABO antibodies-serological behavior and immuno-chemical characterization. J. Immunogenet. 17, 279–99.
Pennington, J. E., Rastan, S., Roelcke, D. & Feizi, T. (1985) Saccharide structures of the mouse embryo during the first eight days of development. J. Embryol. Exp. Morphol. 90, 335–61.
Saito, O., Wang, W.-C., Lotan, R. & Fukuda, M. (1992) Differential glycosylation and cell surface expression of lysosomal membrane glycoproteins in sublines of a human colon cancer exhibiting distinct metastatic potentials. J. Biol. Chem. 267, 5700–11.
Scudder, P., Hanfland, P., Uemura, K. & Feizi, T. (1984) Endo-β-galactosidases of Bacteroides fragilis and Escherichia freundii hydrolyze linear but not branched oligosaccharide domains of glycolipids of the neolacto series. J. Biol. Chem. 259, 6585–92.
Spooncer, E., Fukuda, M., Klock, J. C., Oates, J. E. & Dell, A. (1984) Isolation and characterization of polyfucosylated lactosaminoglycan from human granulocytes. J. Biol. Chem. 259, 4792–801.
Tsay, G. C., Dawson, G. & Li, Y.-T. (1975) Structure of the glycopeptide storage material in GM1 gangliosidosis. Sequence determination with specific endo-and exoglycosidases. Biochim. Biophys. Acta 385, 305–11.
Wang, W.-C., Lee, N., Aoki, D., Fukuda, M. N. & Fukuda, M. (1991) The poly-N-acetyllactosamines attached to lysosomal membrane glycoproteins are increased by the prolonged association with the Golgi complex. J. Biol. Chem. 266, 23 185–90.
Watkins, W. M. (1980) Biochemistry and genetics of the ABO, Lewis, and P blood group systems. In Advances in Human Genetics (edited by Harris, H. & Hirschorn, K.), pp. 1–116. New York: Plenum Press.
Wiederschain, G. Y. & Rosenfeld, R. L. (1971) The forms of -l-fucosidase from pig kidney and their action on natural oligosaccharides. Biochem. Biophys. Res. Commun. 44, 1008–14.
Yamada, K., Shimizu, S. & Takahashi, N. (1983) Histochemical demonstration of asparagine-linked oligosaccharides in glycoproteins of human placenta and umbilical cord tissues by means of almond glycopeptidase digestion. Histochem. J. 15, 1239–50.
Yamashita, K., Ohkura, T., Tachibana, Y., Takasaki, S. & Kobata, A. (1984) Comparative study of the oligosaccharides released from baby hamster kidney cells and their polyoma transformant by hydrazinolysis. J. Biol. Chem. 259, 10 834–40.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ito, N., Kawahara, S., Hirano, Y. et al. Localization of blood-group-related linear poly-N-acetyllactosamine structure in different human tissues by Griffonia simplicifolia agglutinin-II staining following endo-β-galactosidase digestion. Histochem J 26, 327–336 (1994). https://doi.org/10.1007/BF00157766
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00157766