Journal of Cell Science, Vol 110, Issue 22 2807-2818, Copyright © 1997 by Company of Biologists

JOURNAL ARTICLES

Changes in the distribution of LFA-1, catenins, and F-actin during transendothelial migration of monocytes in culture

M Sandig, E Negrou and KA Rogers
Department of Anatomy and Cell Biology, The University of Western Ontario, London, Canada. msandig@julian.uwo.ca

To determine changes in the spatial and temporal distribution of cell-cell adhesion molecules during transendothelial migration of monocytes, we examined an in vitro model system of diapedesis using high resolution laser scanning confocal microscopy. Human arterial endothelial cells were cultured to confluence on coverslips coated with Matrigel and activated with IL-1beta before the addition of monocytic THP-1 cells. Seventy per cent of monocytes transmigrated through the endothelium within one hour. Diapedesis, but not adhesion and spreading, was inhibited 8-fold in co-cultures that contained endothelial cell conditioned medium, suggesting the release of an endothelial derived inhibitor. Double immunofluorescence labeling with antibodies to LFA-1, alpha- and beta-catenin, VE-cadherin and with Texas Red phalloidin, identified a circular transmigration passage in endothelial cell-cell contact regions. This passage was formed by an LFA-1-containing pseudopodium that penetrated between endothelial cells. Apical to the transmigration passage, monocytes remained round in shape, while underneath the endothelium, they spread along the Matrigel. The margins of the transmigration passage contained high levels of LFA-1 and F-actin, suggesting a major role of these molecules during the transmigration process itself. Endothelial adherens junctions, as judged by the presence of VE-cadherin and alpha-catenin adjacent to the passage, remained intact during diapedesis. The presence of catenins at heterotypic contact regions between monocytes and endothelial cells during diapedesis suggested cadherin-mediated interactions between the two cell types. These results reveal dynamic changes in the distribution of adhesion molecules and the actin cytoskeleton during monocyte transendothelial migration in culture.

This article has been cited by other articles:

				S. Lemaire, F. Van Bambeke, M.-P. Mingeot-Leclercq, and P. M. Tulkens Modulation of the Cellular Accumulation and Intracellular Activity of Daptomycin towards Phagocytized Staphylococcus aureus by the P-Glycoprotein (MDR1) Efflux Transporter in Human THP-1 Macrophages and Madin-Darby Canine Kidney Cells Antimicrob. Agents Chemother., August 1, 2007; 51(8): 2748 - 2757. [Abstract] [Full Text] [PDF]

				L. Yang, J. R. Kowalski, X. Zhan, S. M. Thomas, and F. W. Luscinskas Endothelial Cell Cortactin Phosphorylation by Src Contributes to Polymorphonuclear Leukocyte Transmigration In Vitro Circ. Res., February 17, 2006; 98(3): 394 - 402. [Abstract] [Full Text] [PDF]

				C. V. Carman and T. A. Springer A transmigratory cup in leukocyte diapedesis both through individual vascular endothelial cells and between them J. Cell Biol., October 25, 2004; 167(2): 377 - 388. [Abstract] [Full Text] [PDF]

				J. D. van Buul and P. L. Hordijk Signaling in Leukocyte Transendothelial Migration Arterioscler. Thromb. Vasc. Biol., May 1, 2004; 24(5): 824 - 833. [Abstract] [Full Text]

				C. V. Carman, C.-D. Jun, A. Salas, and T. A. Springer Endothelial Cells Proactively Form Microvilli-Like Membrane Projections upon Intercellular Adhesion Molecule 1 Engagement of Leukocyte LFA-1 J. Immunol., December 1, 2003; 171(11): 6135 - 6144. [Abstract] [Full Text] [PDF]

				J. A. Ronald, C. V. Ionescu, K. A. Rogers, and M. Sandig Differential regulation of transendothelial migration of THP-1 cells by ICAM-1/LFA-1 and VCAM-1/VLA-4 J. Leukoc. Biol., October 1, 2001; 70(4): 601 - 609. [Abstract] [Full Text] [PDF]

				W.-H. Su, H.-i. Chen, J.-p. Huang, and C. J. Jen Endothelial [Ca2+]i signaling during transmigration of polymorphonuclear leukocytes Blood, December 1, 2000; 96(12): 3816 - 3822. [Abstract] [Full Text] [PDF]

				A. A. Preobrazhensky, S. Dragan, T. Kawano, M. A. Gavrilin, I. V. Gulina, L. Chakravarty, and P. E. Kolattukudy Monocyte Chemotactic Protein-1 Receptor CCR2B Is a Glycoprotein That Has Tyrosine Sulfation in a Conserved Extracellular N-Terminal Region J. Immunol., November 1, 2000; 165(9): 5295 - 5303. [Abstract] [Full Text] [PDF]

				S. Noria, D. B. Cowan, A. I. Gotlieb, and B. L. Langille Transient and Steady-State Effects of Shear Stress on Endothelial Cell Adherens Junctions Circ. Res., September 17, 1999; 85(6): 504 - 514. [Abstract] [Full Text] [PDF]

				G Cepinskas, M Sandig, and P. Kvietys PAF-induced elastase-dependent neutrophil transendothelial migration is associated with the mobilization of elastase to the neutrophil surface and localization to the migrating front J. Cell Sci., January 6, 1999; 112(12): 1937 - 1945. [Abstract] [PDF]

				E. J. PETTIT Cytosolic Free Calcium and the Cytoskeleton in the Control of Leukocyte Chemotaxis Physiol Rev, October 1, 1998; 78(4): 949 - 967. [Abstract] [Full Text] [PDF]