Influence of asparagine-linked oligosaccharides on tumor cell recognition in the mixed lymphocyte reaction.
The ability of an Ia+ B cell lymphoma, AKTB-1b, to stimulate thymocytes in the allogeneic mixed lymphocyte reaction is dependent on its prior treatment with either swainsonine or deoxynojirimycin, two inhibitors of the processing of asparagine-linked oligosaccharides. In the absence of drug treatment, the tumor cells fail to stimulate thymocytes, whereas pretreatment of the tumor cells with either drug results in a five- to 10-fold increase in their ability to induce thymocyte proliferation. Drug-treated AKTB-1b stimulates thymocytes at levels comparable to those obtained with allogeneic splenocytes. In contrast, the untreated lymphoma does stimulate splenic lymphocytes, and pretreatment with either inhibitor only marginally increases the response. Genetic studies demonstrate that the thymocyte response is still H-2 locus restricted and can be blocked by monoclonal antibodies against two tumor cell major histocompatibility antigens, H-2K and I-A. Drug treatment does not change cell surface I-A expression, and H-2K levels are apparently decreased one-third by deoxynojirimycin but are not affected by swainsonine. To verify that the drug protocol used was capable of altering the glycoconjugates of membrane-associated proteins, the endo-beta-N-acetylglucosaminidase H (endo H)-sensitivity of immunopurified H-2K and I-A was analyzed by SDS-PAGE. These studies demonstrated that swainsonine treatment does result in cell surface expression of glycoconjugates with altered oligosaccharide moieties. Likewise, deoxynojirimycin treatment results in the cell surface expression of an I-A alpha polypeptide with altered oligosaccharide chains while only marginally affecting H-2K and not affecting the I-A beta chain. An intracellular form of the I-A beta chain sensitive to endo H digestion in the presence of deoxynojirimycin is not detectable at the cell surface. Neuraminidase-digested AKTB-1b are also capable of stimulating allogeneic thymocytes. These studies demonstrate that changes in the glycosylation state of the tumor cell can markedly influence its recognition by allogeneic lymphocytes, and further, that different T cell populations differ in their response to such changes.
Powell
L D
LD
Bause
E
E
Legler
G
G
Molyneux
R J
RJ
Hart
G W
GW
D017485:1-Deoxynojirimycin; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D001216:Asparagine; D002236:Carbohydrate Conformation; D002460:Cell Line; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D006183:H-2 Antigens / Q000235:genetics; D008213:Lymphocyte Activation / Q000187:drug effects; D007959:Lymphocyte Culture Test, Mixed; D008223:Lymphoma / Q000276:immunology / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D009363:Neoplasm Proteins / Q000096:biosynthesis; D009439:Neuraminidase / Q000494:pharmacology; D009844:Oligosaccharides / Q000378:metabolism; D013154:Spleen / Q000166:cytology; D017026:Swainsonine; D013601:T-Lymphocytes / Q000276:immunology; D013950:Thymus Gland / Q000166:cytology
D000470:Alkaloids; D000954:Antigens, Surface; D006183:H-2 Antigens; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D009844:Oligosaccharides; D017485:1-Deoxynojirimycin; D001216:Asparagine; D009439:Neuraminidase; D005944:Glucosamine; D017026:Swainsonine