A cloned CD15s-negative variant of HL60 cells is deficient in expression of FUT7 and does not adhere to cytokine-stimulated endothelial cells.
The initial steps of leukocyte adhesion depend on selectin/ligand interactions. Surface ligands on leukocytes are often modified by addition of the sialyl Lewis x (CD15s) determinant. Biosynthesis of CD15s is dependent upon alpha(2,3)sialyltransferases and alpha(1,3)fucosyltransferases. We report the isolation of an HL60 cell line variant, HL60A2, that no longer expresses CD15s. HL60A2 cells do not adhere to cytokine-stimulated endothelial cells. Enzymatic assays reveal that this cell line has normal alpha(2,3)sialyltransferase activity but is deficient in the alpha(1,3)fucosyltransferase responsible for biosynthesis of CD15s (FUT7). The fucosyltransferase that constructs the non-sialylated antigen, Lewis x (CD15), is expressed at high levels (FUT4). Transcript analyses show that FUT7 and FUT4 are inversely expressed in HL60 and variant cell lines. HL60A2 cells provide a tool to study the regulation of selectin ligands and corresponding human fucosyltransferase genes.
The Lineberger Comprehensive Cancer Center and the Department of Pediatrics, University of North Carolina, Chapel Hill 27599-7220, USA. bwwmd@med.unc.edu
B W
BW
Weston
K M
KM
Hiller
J P
JP
Mayben
G
G
Manousos
C M
CM
Nelson
M B
MB
Klein
J L
JL
Goodman
D000818:Animals; D002448:Cell Adhesion; D002999:Clone Cells / Q000201:enzymology; D016207:Cytokines / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D019084:Fluorescent Antibody Technique, Indirect; D005647:Fucosyltransferases / Q000096:biosynthesis*; D015971:Gene Expression Regulation, Enzymologic; D018922:HL-60 Cells; D006801:Humans; D007983:Lewis Blood Group Antigens; D016256:Lewis X Antigen / Q000032:analysis*; D051379:Mice; D009844:Oligosaccharides / Q000032:analysis*; D000080506:Sialyl Lewis X Antigen
D016207:Cytokines; D007983:Lewis Blood Group Antigens; D016256:Lewis X Antigen; D009844:Oligosaccharides; D000080506:Sialyl Lewis X Antigen; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase