Ana Marui (a), Anita Markoti (b), Nataa Kovai (a) and Johannes Müthing (c)
Carbohydrate Research, 2004, 339: 77-86
Abstract
The expression of gangliosides and neutral glycosphingolipids (GSLs) in the lymph nodes of mice lacking the gene for the tumour necrosis factor- receptor p55 (TNFR1) has been investigated. GSL expression in the tissues of mice homozygous (TNFR1-/-) or heterozygous (TNFR1+/-) for the gene deletion was analysed by flow cytometry and high-performance thin-layer chromatography (HPTLC) followed by immunostaining with specific antibodies. HPTLC immunostaining revealed that lymph nodes from TNFR1-/- mice had reduced expression of ganglioside GM1b and GalNAc-GM1b, neolacto-series gangliosides, as well as the globo- (Gb3, Gb4 and Gb5) and ganglio-series (Gg3 and Gg4) neutral GSLs. Flow cytometry of freshly isolated lymph node cells showed no significant differences in GSL expression, except for the GalNAc-GM1b ganglioside, which was less abundant on T lymphocytes from TNFR1-/- lymph nodes. In TNFR1-/- mice, GalNAc-GM1b+/CD4+ T cells were twofold less abundant (3.8% vs 7.6% in the control mice), whereas GalNAc-GM1b+/CD8+ T cells were fourfold less abundant (5.0% vs 20.2% in the control mice). This study provides in vivo evidence that TNF signalling via the TNFR1 is important for the activation of GM1b-type ganglioside biosynthetic pathway in CD8 T lymphocytes, suggesting its possible role in the effector T lymphocyte function.
Graphical Abstract
Expression of gangliosides and neutral glycosphingolipids was investigated in lymph nodes of mice with TNF receptor 1 gene knockout, using HPTLC immunooverlay and flow cytometry. Results show that TNF receptor 1 is important for the expression of GalNAc-GM1b, especially on effector CD8+ T lymphocytes.
Author Keywords: Glycosphingolipids; Gangliosides; Lymph nodes; TNF receptor 1; Knockout mice; HPTLC immunostaining; Flow cytometry Abbreviations: GSL(s), glycosphingolipid(s); HPTLC, high-performance thin-layer chromatography; Neu5Ac, N-acetylneuraminic acid; Neu5Gc, N-glycolylneuraminic acid; PBS, phosphate buffered saline; TNF, tumour necrosis factor; TNFR1, TNF receptor p55; TNFR1-/-, mice homozygous for the TNFR1 gene knockout; TNFR1+/-, mice heterozygous for the TNFR1 gene knockout. Glycosphingolipid nomenclature according to the IUPAC-IUB recommendations: lactosylceramide, Lc2 or LacCer, Gal4Glc1Cer; Globotriaosylceramide or Gb3, Gal4Gal4Glc1Cer; Globotetraosylceramide or Gb4, GalNAc3Gal4Gal4Glc1Cer; Globopentaosylceramide or Gb5, GalNAc3GalNAc3Gal4Gal4Glc1Cer; Gangliotriaosylceramide or Gg3, GalNAc4Gal4Glc1Cer; Gangliotetraosylceramide or Gg4, Gal3GalNAc4Gal4Glc1Cer; Gangliopentaosylceramide or Gg5, GalNAc4Gal3GalNAc4Gal4Glc1Cer; Lacto-N-neotetraosylceramide or nLc4, Gal4GlcNAc3Gal4Glc1Cer; Lacto-N-neohexaosylceramide or nLc6, Gal4GlcNAc3Gal4GlcNAc3Gal4Glc1Cer; GM3, II3Neu5Ac-Lc2; GM2,II3Neu5Ac-Gg3; GM1 or GM1a, II3Neu5Ac-Gg4; GM1b, IV3Neu5Ac-Gg4; GalNAc-GM1b, IV3Neu5Ac-Gg5; GD1, IV3Neu5Ac,III6Neu5Ac-Gg4; GD1a, IV3Neu5Ac, II3Neu5Ac-Gg4; GD1c, IV3(Neu5Ac)2-Gg4 (only Neu5Ac-substituted gangliosides are presented in this list of abbreviations)
Corresponding author. Tel.: +385-1-459-0222.
(a) Institute for Brain Research and Department of Anatomy, Zagreb University School of Medicine, alata 3, HR-10000, Zagreb, Croatia
(b) Department of Biochemistry, Split University School of Medicine, oltanska 2, HR-21000, Split, Croatia
(c) Institute for Medical Physics and Biophysics, University of Münster, Robert-Koch-Str. 31, D-48149, Münster, Germany | 
