Human T lymphocyte isolation, culture and analysis of migration in vitro.
The migration of T lymphocytes involves the adhesive interaction of cell surface integrins with ligands expressed on other cells or with extracellular matrix proteins. The precise spatiotemporal activation of integrins from a low affinity state to a high affinity state at the cell leading edge is important for T lymphocyte migration. Likewise, retraction of the cell trailing edge, or uropod, is a necessary step in maintaining persistent integrin-dependent T lymphocyte motility. Many therapeutic approaches to autoimmune or inflammatory diseases target integrins as a means to inhibit the excessive recruitment and migration of leukocytes. To study the molecular events that regulate human T lymphocyte migration, we have utilized an in vitro system to analyze cell migration on a two-dimensional substrate that mimics the environment that a T lymphocyte encounters during recruitment from the vasculature. T lymphocytes are first isolated from human donors and are then stimulated and cultured for seven to ten days. During the assay, T lymphocytes are allowed to adhere and migrate on a substrate coated with intercellular adhesion molecule-1 (ICAM-1), a ligand for integrin LFA-1, and stromal cell-derived factor-1 (SDF-1). Our data show that T lymphocytes exhibit a migratory velocity of approximately 15 microm/min. T lymphocyte migration can be inhibited by integrin blockade or by inhibitors of the cellular actomyosin machinery that regulates cell migration.
Lefort
Craig T
CT
Center for Vaccine Biology and Immunology, University of Rochester, USA.
Kim
Minsoo
M
D054441:Cell Migration Assays, Leukocyte / Q000379:methods*; D002465:Cell Movement / Q000502:physiology*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000097:blood; D016169:Lymphocyte Function-Associated Antigen-1 / Q000097:blood; D013601:T-Lymphocytes / Q000166:cytology*
D016169:Lymphocyte Function-Associated Antigen-1; D018799:Intercellular Adhesion Molecule-1