title: Expression of gangliosides GM3 (NeuAc) and GM3 (NeuGc) in myelomas and hybridomas of mouse, rat, and human origin.
abstract: In this study gangliosides from various myelomas and hybridomas of mouse, rat, and human origin were characterized by thin-layer and high-performance liquid chromatography, immunological methods (overlay technique) and fast atom bombardment mass spectrometry. Exclusively GM3 substituted with C24:1- and C16:0-fatty acid, was found in all B cell-derived cell lines. C18 sphingosine was the single long chain base in each GM3 ceramide portion. The mouse myeloma (NS-1) and all hybridomas, obtained by fusion of mouse, rat, or human B lymphocytes with murine myelomas, showed high GM3 (NeuGc) content (> 75%) and low GM3 (NeuAc) expression. Absolute amounts of GM3 ranged from 0.2 up to 0.8 mg x 10(-9) cells. Normally, human cells do not express NeuGc, and an Epstein-Barr virus-transformed human B lymphocyte line analyzed in this study retained this sialylation status, expressing exclusively GM3 (NeuAc) (100%). The fusion of human B lymphocytes with mouse myelomas led to high GM3 (NeuGc) expression (average about 85%) in all mouse/human heterohybridomas examined. Our results indicate the chromosomal gene "transfer" and/or the activation of enzymes involved in NeuGc-biosynthesis due to the somatic cell fusion process, which might explain the mouse dominance in the manifestation of the NeuGc-phenotype in hybridomas of human origin.
authors.affiliation: Institut für Zellkulturtechnik der Universität, Bielefeld, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Müthing
authors.forename: H
authors.initials: H
authors.lastname: Steuer
authors.forename: J
authors.initials: J
authors.lastname: Peter-Katalinić
authors.forename: U
authors.initials: U
authors.lastname: Marx
authors.forename: U
authors.initials: U
authors.lastname: Bethke
authors.forename: U
authors.initials: U
authors.lastname: Neumann
authors.forename: J
authors.initials: J
authors.lastname: Lehmann
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D002855:Chromatography, Thin Layer; D005679:G(M3) Ganglioside / Q000032:analysis*; D006801:Humans; D006825:Hybridomas / Q000737:chemistry*; D051379:Mice; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000378:metabolism*; D019158:N-Acetylneuraminic Acid; D051381:Rats; D012794:Sialic Acids / Q000032:analysis; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured
chemical_list: D005679:G(M3) Ganglioside; D012794:Sialic Acids; D019158:N-Acetylneuraminic Acid