Identification of CD66a and CD66b as the major galectin-3 receptor candidates in human neutrophils.
The mammalian lectin galectin-3 is a potent stimulus of human neutrophils, provided that the receptor(s) for the lectin has been mobilized to the cell surface before activation. We have recently shown that the receptors for galectin-3 are stored in intracellular mobilizable granules. Here we show supportive evidence for this in that DMSO-differentiated (neutrophil-like) HL-60 cells, which lack gelatinase and specific granules, are nonresponsive when exposed to galectin-3. Neutrophil granules were subsequently used for isolation of galectin-3 receptors by affinity chromatography. Proteins eluted from a galectin-3-Sepharose column by lactose were analyzed on SDS-polyacrylamide gels and showed two major bands of 100 and 160 kDa and a minor band of 120 kDa. By immunoblotting, these proteins were shown to correspond to CD66a (160 kDa), CD66b (100 kDa), and lysosome-associated membrane glycoprotein-1 and -2 (Lamp-1 and -2; 120 kDa). The unresponsive HL-60 cells lacked the CD66 Ags but contained the Lamps, implying that neutrophil CD66a and/or CD66b may be the functional galectin-3 receptors. This conclusion was supported by the subcellular localization of the CD66 proteins to the gelatinase and specific granules in resting neutrophils.
Feuk-Lagerstedt
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The Phagocyte Research Laboratory, Department of Medical Microbiology and Immunology, University of Göteborg, Sweden.
Jordan
E T
ET
Leffler
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H
Dahlgren
C
C
Karlsson
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A
D015703:Antigens, CD / Q000096:biosynthesis / Q000097:blood* / Q000378:metabolism; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000097:blood* / Q000378:metabolism / Q000494:pharmacology; D002352:Carrier Proteins / Q000097:blood; D015815:Cell Adhesion Molecules; D002454:Cell Differentiation / Q000187:drug effects; D003594:Cytoplasmic Granules / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D037502:Galectin 3; D018093:Gelatinases / Q000097:blood; D018922:HL-60 Cells; D006801:Humans; D051907:Lysosomal Membrane Proteins; D016177:Macrophage-1 Antigen / Q000096:biosynthesis; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D018375:Neutrophil Activation / Q000276:immunology; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000096:biosynthesis / Q000097:blood* / Q000378:metabolism; D013347:Subcellular Fractions / Q000276:immunology / Q000378:metabolism
D015703:Antigens, CD; D000943:Antigens, Differentiation; C069847:CD66 antigens; D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D037502:Galectin 3; D051907:Lysosomal Membrane Proteins; D016177:Macrophage-1 Antigen; D008562:Membrane Glycoproteins; D011975:Receptors, Mitogen; D018093:Gelatinases; D004121:Dimethyl Sulfoxide