Characterization of the E-selectin ligand on NK cells.
In this study we demonstrate that human CD56+CD16+/CD3- NK cells adhere to the E-selectin expressed by stimulated HUVEC in a sialidase- and Ca(2+)-dependent manner, and express a silylated Lex adhesion structure. We have characterized this sLe(x) epitope on NK cell in detail and show here that the sLe(x) on NK cells was not recognized by the CSLEX1 Ab, but was readily identified by two anti-di-sLe(x) Abs, KM-93 and FH-6. Furthermore, cleaving sialic acid with a sialidase treatment revealed a pool of Le(x) epitopes on the NK cells surface, providing further proof that NK cells express sLe(x) epitopes. Extensive protease treatments did not cleave the sLe(x) epitope from NK cells, which suggests that it could be linked to a lipid backbone. This di-sLe(x) was able to mediate adhesion to E-selectin, suggesting that it represents an essential part or is closely related to a selectin ligand on NK cells. We were also able to show that NK cells possess several alpha 2,3 sialyltransferases and alpha 1,3 or alpha 1,3/4 fucosyltransferases. These enzymes are crucial in the synthesis of sLe(x) epitopes on cell surfaces. Taken together, we provide evidence that NK cells have a di-sLe(x) oligosaccharide capable of adhesion to E-selectin, and NK cells have the machinery (i.e., relevant transferases) to generate these sialylated Lewis oligosaccharides.
Pinola
M
M
Department of Pathology, University of Helsinki, Finland.
Renkonen
R
R
Majuri
M L
ML
Tiisala
S
S
Saksela
E
E
D002118:Calcium / Q000378:metabolism; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000378:metabolism; D005647:Fucosyltransferases / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D007983:Lewis Blood Group Antigens / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D008024:Ligands; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D017452:Receptors, IgG / Q000032:analysis; D012799:Sialyltransferases / Q000378:metabolism
D015815:Cell Adhesion Molecules; D019040:E-Selectin; D007983:Lewis Blood Group Antigens; D008024:Ligands; D012333:RNA, Messenger; D017452:Receptors, IgG; D005647:Fucosyltransferases; D012799:Sialyltransferases; D009439:Neuraminidase; D002118:Calcium