Structural characterization of gangliosides from murine T lymphocytes.
Mouse spleen cells were prepared from CBA/J mice, and T lymphocytes were selectively stimulated with the T cell mitogen concanavalin A and further propagated in the presence of the T cell growth factor interleukin-2. The T cells were metabolically labeled with D-[1-14C]galactose and D[1-14C]glucosamine, and the gangliosides were extracted and purified by DEAE-Sepharose column chromatography. Carbohydrate backbone structures of the asialogangliosides, prepared by mild acid hydrolysis, were determined by high-performance liquid chromatography, treatment with exoglycosidases and immunostaining. Monosialylated gangliosides were isolated by gradient elution from DEAE-Sepharose and further separated by preparative high-performance thin-layer chromatography in two solvent systems. Isolated fractions were characterized by preparation of asialogangliosides by mild acid hydrolysis, the action of Vibrio cholerae neuraminidase, and fast-atombombardment mass spectrometry. The following structures were identified: IVNeuAc-GgOse4Cer; IVNeuGc-GgOse4Cer; IVNeuAc-GgOse5Cer; and IVNeu-Gc-GgOse5Cer. The latter two gangliosides were not detected on B lymphoblasts and may be T-cell-specific structures. All gangliosides were heterogeneous in their ceramide moieties, being substituted with C16:0, C24:0, and C24:1 fatty acids. A preliminary study of several other mouse strains showed no strain-specific genetic variations in the T cell gangliosides. The possible role of these gangliosides is discussed.
D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer / Q000379:methods; D003208:Concanavalin A / Q000494:pharmacology; D005260:Female; D005732:Gangliosides / Q000097:blood*; D006028:Glycosphingolipids / Q000097:blood; D006868:Hydrolysis; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008808:Mice, Inbred CBA; D013601:T-Lymphocytes / Q000032:analysis* / Q000187:drug effects
D005732:Gangliosides; D006028:Glycosphingolipids; D003208:Concanavalin A