Concanavalin A-stimulated expression of gangliosides with GalNAc beta 1-4(NeuAc alpha 2-3)Gal beta structure in murine thymocytes.
We analysed the glycolipids of mouse thymocytes before and after Concanavalin A (Con A) or recombinant interleukin-2 (rIL-2) stimulation by TLC-immunostaining with carbohydrate-specific antiglycolipid antibodies. The thymocytes were cultured in serum-free medium in the presence of 500 ng ml-1 Con A, 10 U ml-1 rIL-2 or Con A plus rIL-2 for 6, 12, 24, 48, and 72 h, and were found to start proliferating 24 h after cultivation in the presence of Con A or Con A plus rIL-2, the maximum levels being reached at 72 h and 48 h, respectively, in a thymidine uptake experiment. The concentrations of II3Neu-Gg4Cer, Gg4Cer and IV3GalNAc alpha-Gb4Cer after 48 h Con A stimulation were found to be at almost the original levels. Conversely, II3Neu-Gg3Cer, which was not detected in the thymocytes at the start, began to appear after 48 h stimulation with Con A and Con A plus rIL-2, and IV3Neu-Gg5Cer in the cells 48 h after stimulation with Con A and Con A plus rIL-2 has increased to 41 and 44 times higher than in the original cells, respectively, as judged on TLC-immunostaining with monoclonal antibody YHD-06, which detects the GalNAc beta 1-4(NeuAc or NeuGc alpha 2-3)Gal beta-structure. These results indicate that the increased synthesis of both gangliosides, in other words, the activation of N-acetylgalactosaminyltransferase, is associated with the mitogen-induced proliferation.(ABSTRACT TRUNCATED AT 250 WORDS)
Horikawa
K
K
Department of Biochemistry, Faculty of Medicine, University of Tokyo, Japan.
Yamasaki
M
M
Iwamori
M
M
Nakakuma
H
H
Takatsuki
K
K
Nagai
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002455:Cell Division; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology*; D000939:Epitopes / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D005732:Gangliosides / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D007376:Interleukin-2 / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011994:Recombinant Proteins / Q000494:pharmacology; D013950:Thymus Gland / Q000166:cytology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005732:Gangliosides; D007376:Interleukin-2; D011994:Recombinant Proteins; D003208:Concanavalin A