Lymph node macrophages, but not spleen macrophages, express high levels of unmasked sialoadhesin: implication for the adhesive properties of macrophages in vivo.
Sialoadhesin is a macrophage-restricted adhesion molecule that recognizes N-acetylneuraminylalpha2-3galactose structure. We prepared a multivalent neoglycoprotein probe carrying this oligosaccharide and characterized the binding activity of sialoadhesin on native rat macrophages. Macrophages from mesenteric and axillar lymph nodes exhibited 36-fold higher activity than those from the spleen. The K(d) values of the probe binding to macrophages of the two organs were indistinguishable (1-2 nM), whereas the B(max) value of lymph node macrophages was markedly higher than that of splenic macrophages. Western blot analysis revealed that the quantity of sialoadhesin present in lymph node macrophages was 25-fold higher than in splenic macrophages. High cell surface expression of sialoadhesin on lymph node macrophages was also shown by flow cytometry. To examine the "masking" of sialoadhesin by endogenous sialoglycoconjugates, we treated macrophages with sialidase before measuring the probe binding. After sialidase treatment, the binding activity of splenic macrophages increased fourfold, whereas that of lymph node macrophages did not increase. In conclusion, we have identified macrophages expressing high levels of unmasked sialoadhesin in lymph nodes. The unmasked forms on these macrophages are available for sialoadhesin-dependent adhesive functions, unlike the masked forms on the majority of splenic macrophages.
Nakamura
Kyoko
K
Glyco-chain Functions Laboratory, Supra-biomolecular System Group, Frontier Research System, Institute of Physical and Chemical Research (RIKEN), Saitama, 351-0198, Japan.
Yamaji
Toshiyuki
T
Crocker
Paul R
PR
Suzuki
Akemi
A
Hashimoto
Yasuhiro
Y
D000818:Animals; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000235:genetics; D004912:Erythrocytes / Q000502:physiology; D007700:Kinetics; D008198:Lymph Nodes; D008264:Macrophages / Q000502:physiology*; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry; D009928:Organ Specificity; D051381:Rats; D017208:Rats, Wistar; D011971:Receptors, Immunologic / Q000378:metabolism*; D012756:Sheep; D063267:Sialic Acid Binding Ig-like Lectin 1; D013154:Spleen
D015815:Cell Adhesion Molecules; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011971:Receptors, Immunologic; D063267:Sialic Acid Binding Ig-like Lectin 1