title: Live imaging of LFA-1-dependent T-cell motility and stop signals.
abstract: T-cell motility is critical for leukocyte trafficking both in normal host defense and in pathologic conditions including chronic inflammatory disease. Despite progress in understanding the mechanisms of T-cell polarity and motility, we have limited understanding of the mechanisms that contribute to antigen-induced T cell arrest. Here, we describe methods to analyze leukocyte function antigen-1-mediated T-cell motility and T-cell receptor-induced stop signals using in vitro assays on two-dimensional surfaces. Specifically, methods for live time-lapse imaging of T cell random migration and arrest on ICAM-1-coated surfaces are described. Additionally, we detail methods for live imaging of T-cell motility within 3D substrates to analyze T cell-antigen-presenting cell (APC) interactions and APC-mediated stop signals.
authors: Department of Medical Microbiology and Immunology, University of Wisconsin, Madison, WI, USA.
authors: Andrew J
authors: AJ
authors: Wiemer
authors: Sarah
authors: S
authors: Wernimont
authors: Anna
authors: A
authors: Huttenlocher
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D000941:Antigens / Q000276:immunology; D002460:Cell Line; D002465:Cell Movement / Q000276:immunology*; D058948:Cell Tracking / Q000379:methods*; D002478:Cells, Cultured; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology / Q000378:metabolism; D016169:Lymphocyte Function-Associated Antigen-1 / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008853:Microscopy; D013601:T-Lymphocytes / Q000276:immunology*; D059008:Time-Lapse Imaging
chemical_list: D000941:Antigens; D016169:Lymphocyte Function-Associated Antigen-1; D018799:Intercellular Adhesion Molecule-1