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Establishment of cells exhibiting mutated glycolipid synthesis from mouse thymus by immortalization with SV-40 virus

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Abstract

Immortalization with simian virus-40 and cloning of immortalized cells from mouse thymus were performed to establish cell lines for characterization of the mode of glycolipid expression in the thymic cells. Among the 25 cell lines obtained, three lines with different morphologies were established, that is, epithelial (IMTH-E), fibroblastic (IMTH-F), and asterisk-like (IMTH-I) cells, and their glycolipids, together with those in the thymus, were determined systematically. The major glycolipids in mouse thymus were the globo- and ganglio-series, both of which, were co-expressed in the three cell lines established. However, the mode of modification of the globo- and ganglio-series was distinct for each cell line. As to the globo-series, the structures with the longest carbohydrate chain for IMTH-E, -F, and -I cells were Gb3Cer, Gb4Cer, and Forssman antigen, respectively, having stepwise shorter carbohydrates at the nonreducing termini. Although the acidic glycolipids in IMTH-E cells comprised GM3 and GM2, and their sulfated isomers, IMTH-F and -I cells expressed GMlb and GDlc for the α-pathway, and up to GDI a for the a-pathway of ganglio-series glycolipids. GMlb-GalNAc present in the thymus was not detected in IMTH-F and -I cells, probably due to the lower synthetic activity for the metabolic intermediate Gg4Cer. The results indicate that the immortalization technique is useful for obtaining individual cells having unique glycolipid profiles for analysis of the functional significance and metabolism of glycolipids in the thymus.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Science and Technology, Kinki University, 3-4-1 Kowakae, Higashiosaka, Osaka, 577-8502, Japan

    Masao Iwamori & Yuriko Iwamori

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  1. Masao Iwamori
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  2. Yuriko Iwamori
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Correspondence to Masao Iwamori.

Additional information

Abbreviations: Glycolipids are abbreviated according to the recommendations of the IUPAC-IUBMB Commission on Biochemical Nomenclature. Eur J Biochem 79, 11-21 (1977). The ganglioside nomenclature of Svennerholm [1] is employed throughout, except that GMlb, GMlb-GalNAc, GMlb-GaINAc-Gal, GDlc, and GDI α denote IV3 NeuAcα-Gg4Cer, IV4GalNAcβ, IV3NeuAcα-Gg4Cer, IV4(Galβ-3GalNAcβ),IV3NeuAcα-Gg4Cer, IV3NeuAcα2-Gg4Cer, and III6 NeuAcα, IV3NeuAcα-Gg4Cer, respectively. BSA, bovine serum albumin; PBS, phosphate-buffered saline; FCS, fetal calf serum; SV, simian virus; FABMS, fast atom bombardment mass spectrometry; NeuAc, N-acetylneuraminic acid; NeuGc, N-glycolylneuraminic acid; Hex, hexose: HexNAc, N-acetylhexosamine.

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Iwamori, M., Iwamori, Y. Establishment of cells exhibiting mutated glycolipid synthesis from mouse thymus by immortalization with SV-40 virus. Glycoconj J 22, 417–425 (2005). https://doi.org/10.1007/s10719-005-4086-8

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  • DOI: https://doi.org/10.1007/s10719-005-4086-8

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