During inflammation, leukocytes bind to the adhesion receptors ICAM-1 and VCAM-1 on the endothelial surface before undergoing transendothelial migration, also called diapedesis. ICAM-1 is also involved in transendothelial migration, independently of its role in adhesion, but the molecular basis of this function is poorly understood. Here we demonstrate that, following clustering, apical ICAM-1 translocated to caveolin-rich membrane domains close to the ends of actin stress fibres. In these F-actin-rich areas, ICAM-1 was internalized and transcytosed to the basal plasma membrane through caveolae. Human T-lymphocytes extended pseudopodia into endothelial cells in caveolin- and F-actin-enriched areas, induced local translocation of ICAM-1 and caveolin-1 to the endothelial basal membrane and transmigrated through transcellular passages formed by a ring of F-actin and caveolae. Reduction of caveolin-1 levels using RNA interference (RNAi) specifically decreased lymphocyte transcellular transmigration. We propose that the translocation of ICAM-1 to caveola- and F-actin-rich domains links the sequential steps of lymphocyte adhesion and transendothelial migration and facilitates lymphocyte migration through endothelial cells from capillaries into surrounding tissue.
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This work was supported by the Ludwig Institute for Cancer Research and European Community contracts QLG1-CT-99-01036 and FP6–502935. J. Millán was supported by a Marie Curie fellowship (no. HPMF-CT-2000-01061) and British Heart Foundation intermediate fellowship (no. FS/04/006). We are grateful to the named donors for the gifts of plasmids and antibodies listed in the methods section, to E. Cernuda Morollon for providing T-lymphoblasts, and to members of the Ridley laboratory for helpful discussions. We thank Olympus for generously providing instrumentation and support to the Yale CINEMA lab.
The authors declare no competing financial interests.
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