N-linked carbohydrate on human leukocyte antigen-C and recognition by natural killer cell inhibitory receptors.
The possible role of carbohydrate in the interaction of HLA-C with a human inhibitory natural Killer cell Immunoglobulin-like Receptor with two Ig domains, KIR2DL1, was investigated. Transfectants of 721.221 (a class I MHC-negative human B cell line) expressing only HLA-Cw4 or -Cw6 or their respective non-glycosylated mutants (N86Q, S88A) were made. The binding of a KIR2DL1-Ig fusion protein to the non-glycosylated mutant HLA-Cw4- or -Cw6-expressing cells was markedly decreased compared to the wild type-expressing cells. The ability to induce an inhibitory signal in the NK tumor line YTS transfected with KIR2DL1 was also impaired in the nonglycosylated mutant expressing cells. Furthermore, in a second functional assay, mutant HLA-Cw4 and -Cw6 molecules had impaired ability to induce signal transduction in BW cells expressing a KIR2DL1-CD3 zeta chain chimeric protein. Thus, the deletion of the N-linked glycosylation signal in HLA-Cw4 and -Cw6 greatly reduced recognition by KIR2DL1. Alternative interpretations of the data are discussed.
Baba
E
E
Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
Erskine
R
R
Boyson
J E
JE
Cohen
G B
GB
Davis
D M
DM
Malik
P
P
Mandelboim
O
O
Reyburn
H T
HT
Strominger
J L
JL
D019943:Amino Acid Substitution / Q000235:genetics / Q000276:immunology; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D001216:Asparagine / Q000235:genetics; D019556:COS Cells; D002236:Carbohydrate Conformation / Q000187:drug effects; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000037:antagonists & inhibitors; D002461:Cell Line, Transformed; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic / Q000187:drug effects / Q000235:genetics; D005973:Glutamine / Q000235:genetics; D006031:Glycosylation / Q000187:drug effects; D015236:HLA-C Antigens / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics; D007694:Killer Cells, Natural / Q000276:immunology / Q000378:metabolism*; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D051920:NK Cell Lectin-Like Receptor Subfamily D; D011485:Protein Binding / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D011971:Receptors, Immunologic / Q000235:genetics / Q000378:metabolism*; D054340:Receptors, KIR; D054342:Receptors, KIR2DL1; D055607:Receptors, Natural Killer Cell; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D017026:Swainsonine / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D002241:Carbohydrates; D015236:HLA-C Antigens; C039966:HLA-C*04 antigen; C528994:HLA-C*06 antigen; D007136:Immunoglobulins; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D051920:NK Cell Lectin-Like Receptor Subfamily D; D011971:Receptors, Immunologic; D054340:Receptors, KIR; D054342:Receptors, KIR2DL1; D055607:Receptors, Natural Killer Cell; D011993:Recombinant Fusion Proteins; D005973:Glutamine; D001216:Asparagine; D017026:Swainsonine