title: Differentially regulated expression of 9-O-acetyl GD3 (CD60b) and 7-O-acetyl-GD3 (CD60c) during differentiation and maturation of human T and B lymphocytes.
abstract: GD3 (CD60a) and its 9-O-acetylated variant (CD60b) are intracellular regulators of apoptosis in T lymphocytes. Surface expressed 9-O-acetyl- and 7-O-acetyl-GD3 (CD60b and CD60c) may have a functional impact on activated T and B cells. In order to investigate the balance between surface and intracellular expression and synthesis and degradation of these glycosphingolipids in human lymphocytes of various differentiation stages, we analyzed (i) expression of GD3 molecules on native T and B cells and thymocytes by flow cytometry and (ii) activity and regulation of possible key enzymes for CD60a,b,c synthesis and degradation at the transcriptional level. Both, surface and cytoplasmic expression of CD60a and CD60c was highest in tonsillar T cells. In thymocytes, CD60c outweighs the other CD60 variants and was mainly found in the cytoplasm. All lymphocyte preparations contained sialate O-acetyltransferase activity producing 7-O-acetyl-GD3. Sialidase activity was highest in peripheral blood lymphocytes followed by thymocytes and tonsillar T and B cells. Transcription of GD3 synthase (ST8SiaI), the key enzyme for GD3 synthesis, was highest in tonsillar T cells, whereas transcriptional levels of sialidase NEU3 and O-acetylesterase H-Lse were lowest in activated T cells. This balance between enzymes of sialic acid metabolism may explain the strong overall staining intensity for all GD3 forms in T cells. Both CASD1, presumably encoding a sialic acid-specific O-acetyltransferase, and H-Lse showed highest transcription in peripheral B lymphocytes corresponding to the low expression of CD60b and c in these cells. Our data point to regulatory functions of these anabolic and catabolic key enzymes for the expression of GD3 and its O-acetylated variants in lymphocytes at a given differentiation stage.
authors.affiliation: German Cancer Research Center, D015 Translational Immunology, Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Wipfler
authors.forename: G Vinayaga
authors.initials: GV
authors.lastname: Srinivasan
authors.forename: Haneen
authors.initials: H
authors.lastname: Sadick
authors.forename: Bernhard
authors.initials: B
authors.lastname: Kniep
authors.forename: Sigrid
authors.initials: S
authors.lastname: Arming
authors.forename: Martina
authors.initials: M
authors.lastname: Willhauck-Fleckenstein
authors.forename: Reinhard
authors.initials: R
authors.lastname: Vlasak
authors.forename: Roland
authors.initials: R
authors.lastname: Schauer
authors.forename: Reinhard
authors.initials: R
authors.lastname: Schwartz-Albiez
mesh_terms: D000107:Acetylation; D000115:Acetylesterase / Q000235:genetics / Q000378:metabolism; D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis / Q000235:genetics / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D002454:Cell Differentiation / Q000276:immunology; D003600:Cytosol / Q000378:metabolism; D005434:Flow Cytometry; D005732:Gangliosides / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000276:immunology*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D009439:Neuraminidase / Q000235:genetics / Q000378:metabolism; D014066:Palatine Tonsil / Q000166:cytology / Q000276:immunology; D012799:Sialyltransferases / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology; D014158:Transcription, Genetic
chemical_list: D000115:Acetylesterase; D000123:Acetyltransferases; D005732:Gangliosides; D008565:Membrane Proteins; D009439:Neuraminidase; D012799:Sialyltransferases; C097213:7-O-acetyl-GD3 ganglioside; C049132:9-O-acetyl-GD3 ganglioside; C561597:CASD1 protein, human; C043662:alpha-N-acetylneuraminate alpha-2,8-sialyltransferase