cytochalasin-d and Leukemia--T-Cell

Integrin-mediated encounters of T cells with extracellular cues lead these cells to adhere to a variety of substrates and acquire a spread phenotype needed for their tissue incursions. We studied the effects of galectin-8 (Gal-8), a beta-galactoside binding lectin, on Jurkat T cells. Immobilized Gal-8 bound alpha1beta1, alpha3beta1 and alpha5beta1 but not alpha2beta1 and alpha4beta1 and adhered these cells with similar kinetics to immobilized fibronectin (FN). Function-blocking experiments with monoclonal anti-integrin antibodies suggested that alpha5beta1 is the main mediator of cell adhesion to this lectin. Gal-8, but not FN, induced extensive cell spreading frequently leading to a polarized phenotype characterized by an asymmetric lamellipodial protrusion. These morphological changes involved actin cytoskeletal rearrangements controlled by PI3K, Rac-1 and ERK1/2 activity. Gal-8-induced Rac-1 activation and binding to alpha1 and alpha5 integrins have not been described in any other cellular system. Strikingly, Gal-8 was also a strong stimulus on Jurkat cells in suspension, triggering ERK1/2 activation that in most adherent cells is instead dependent on cell attachment. In addition, we found that patients with systemic lupus erythematosus (SLE), a prototypic autoimmune disorder, produce Gal-8 autoantibodies that impede both its binding to integrins and cell adhesion. These are the first function-blocking autoantibodies reported for a member of the galectin family. These results indicate that Gal-8 constitutes a novel extracellular stimulus for T cells, able to bind specific beta1 integrins and to trigger signaling pathways conducive to cell spreading. Gal-8 could modulate a wide range of T cell-driven immune processes that eventually become altered in autoimmune disorders.

Topics: Androstadienes; Antibodies, Monoclonal; Autoantibodies; Cell Adhesion; Cell Shape; Cell Surface Extensions; Cytochalasin D; Cytoskeleton; Fibronectins; Flavonoids; Galectins; Humans; Integrin beta1; Jurkat Cells; Leukemia, T-Cell; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Phosphoinositide-3 Kinase Inhibitors; Protein Binding; Protein Kinase Inhibitors; rac1 GTP-Binding Protein; Thiogalactosides; Transfection; Wortmannin