galactocerebroside and Glioma

Expression of galactocerebroside (GalC) was detected in human glioma cell line (U-87 MG). Exposure of cells to antibody against GalC and fluoresceinated second antibody showed intense fluorescence on the plasma membrane. Possible involvement of GalC in receptor-mediated transmembrane signaling was explored in this cell line. Antibodies raised against GalC were used as ligands. Binding of anti-GalC to these cells caused a transient increase in intracellular free calcium ([Ca2+]i). The response was observed both in the presence and absence of extracellular calcium demonstrating that the rise in [Ca2+]i induced by anti-GalC was due to an influx of Ca2+ through plasma membrane as well as the release of Ca2+ from intracellular pools. Ca2+ influx was blocked by verapamil, indicating that influx is mediated by voltage-sensitive channels. Our results suggest that GalC can play a role in transmembrane signaling by modulation of voltage-sensitive Ca2+ channels.

Topics: Calcium; Calcium Channels; Galactosylceramides; Glioma; Humans; Immunohistochemistry; Signal Transduction; Tumor Cells, Cultured

Human immunodeficiency virus (HIV-1) infection in the human brain leads to characteristic neuropathological changes, which may result indirectly from interactions of the envelope glycoprotein gp120 with neurons and/or glial cells. We therefore investigated the binding of recombinant gp120 (rgp120) to human neural cells and its effect on intracellular signalling. Here we present evidence that rgp120, besides binding to galactocerebroside or galactosyl-sulfatide, specifically binds to a protein receptor of a relative molecular mass of approximately 180,000 Da (180 kDa) present on the CD4-negative glioma cells D-54, but not on Molt4 T lymphocytes. Binding of rgp120 to this receptor rapidly induced a tyrosine-specific protein kinase activity leading to tyrosine phosphorylation of 130- and 115-kDa proteins. The concentration of intracellular calcium was not affected by rgp120 in these cells. Our data suggest a novel signal transducing HIV-1 gp120 receptor on CD4-negative glial cells, which may contribute to the neuropathological changes observed in HIV-1-infected brains.

Topics: Brain; Calcium; CD4 Antigens; CD4-Positive T-Lymphocytes; Cell Communication; Cell Membrane; Cross-Linking Reagents; Enzyme Activation; Galactosylceramides; Glioma; HIV Envelope Protein gp120; Humans; Phosphorylation; Protein-Tyrosine Kinases; Recombinant Proteins; Signal Transduction; Sulfoglycosphingolipids; Tumor Cells, Cultured