Comparison of E-selectin-binding glycoprotein ligands on human lymphocytes, neutrophils, and bovine gamma delta T cells.
We compared E-selectin-binding cell surface ligands on bovine gamma delta T cells and human leukocytes using an E-selectin/Ig chimera. The chimera worked well in flow cytometric studies and showed that bovine gamma delta T cells were the only lymphocyte population in newborn animals that bound E-selectin chimera. Furthermore, the chimera blocked gamma delta T cell rolling on E-selectin. Chimera reacted with four potential glycoprotein ligands of 180, 200, 250, and 300 kDa in Western blot analysis of gamma delta T cell detergent lysates, and it specifically precipitated at least two of these E-selectin ligands (200 and 250 kDa) from lysates of cell surface biotinylated gamma delta T cells. Preclearing bovine gamma delta lysates of GD3.5 Ag and workshop cluster 1 did not abrogate E-selectin ligand precipitation, suggesting that these surface markers do not represent E-selectin ligands. Human neutrophils possessed three E-selectin-binding ligands of approximately 80 to 90, 130, and 230 kDa, while human lymphocytes variably possessed three ligands of 120, approximately 220 to 240, and 260 kDa. Cross-precipitation experiments confirmed the results of others that neutrophil L-selectin serves as the 80 to 90-kDa E-selectin ligand. The human lymphocyte approximately 220 to 240-kDa and 260-kDa ligands may be analogous to the bovine gamma delta T cell molecules, whereas the 120-kDa is unique to human cells. The identities of the human and bovine lymphocyte E-selectin ligands are unknown. Finally, E-selectin ligand-1 apparently may have a minimal role, if any, in lymphocyte/E-selectin interactions, since a polyclonal anti-E-selectin ligand-1 serum stained a minimal number of cutaneous lymphocyte-associated Ag-positive human lymphocytes and bovine gamma delta T cells.
Jones
W M
WM
Department of Veterinary Molecular Biology, Montana State University, Bozeman 59717, USA.
Watts
G M
GM
Robinson
M K
MK
Vestweber
D
D
Jutila
M A
MA
D000818:Animals; D000954:Antigens, Surface / Q000097:blood; D015153:Blotting, Western; D002417:Cattle; D015815:Cell Adhesion Molecules / Q000302:isolation & purification / Q000378:metabolism*; D019040:E-Selectin / Q000097:blood* / Q000302:isolation & purification; D018984:Epitopes, T-Lymphocyte / Q000097:blood; D005434:Flow Cytometry; D006023:Glycoproteins / Q000097:blood* / Q000302:isolation & purification; D006801:Humans; D019041:L-Selectin / Q000502:physiology; D008024:Ligands; D008562:Membrane Glycoproteins / Q000502:physiology; D009504:Neutrophils / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding / Q000276:immunology; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000097:blood*; D017468:Receptors, Fibroblast Growth Factor / Q000502:physiology; D016201:Receptors, Lymphocyte Homing / Q000378:metabolism*; D012795:Sialoglycoproteins; D016176:T-Lymphocyte Subsets / Q000378:metabolism*
D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D018984:Epitopes, T-Lymphocyte; D006023:Glycoproteins; D008024:Ligands; D008562:Membrane Glycoproteins; D016692:Receptors, Antigen, T-Cell, gamma-delta; D017468:Receptors, Fibroblast Growth Factor; D016201:Receptors, Lymphocyte Homing; D012795:Sialoglycoproteins; C086711:cysteine-rich fibroblast growth factor receptor; D019041:L-Selectin