Adhesion, transendothelial migration, and reverse transmigration of in vitro cultured dendritic cells

Giovanna D'Amico, Giancarlo Bianchi, Sergio Bernasconi, Laura Bersani, Lorenzo Piemonti, Silvano Sozzani, Alberto Mantovani, Paola Allavena

Research output: Contribution to journalArticlepeer-review


Dendritic cells (DC) are migratory cells which exhibit complex trafficking properties in vivo, involving interaction with vascular and lymphatic endothelium end extracellular matrix (ECM). The underlying mechanisms involved in these processes are still ill defined. In the present study we have investigated the ability of DC to interact in vitro with human vascular endothelial cells (EC) and ECM. DC were differentiated from monocytes by in vitro exposure to granulocyte-macrophage colony-stimulating factor and interleukin-13 for 7 days. In adhesion assays a considerable proportion of DC bound to resting EC monolayers: (17% ± 4%, mean ± SE of eight experiments). Adhesion to tumor necrosis factor (TNF)activated EC was increased to 29% ± 5% (n = 8). Binding to resting EC was strongly inhibited by anti-CD11a and CD11b, but not by CD11c monoclonal antibodies (MoAbs); on TNF-activated EC, anti-VLA-4 in concert with anti-CD18 inhibited adhesion by more than 70%. Binding to a natural ECM, derived from cultured EC, or to purified fibronectin was high: 52% ± 6% (n = 8) involved VLA-4 and VLA-5 integrins. In s transmigration assay, 10% ± 2% (n = 6) of input cells were able to cross the EC monolayer. Unlike adhesion, transendothelial migration was significantly reduced by anti-CD31 MoAb. The amount of DC transmigrated through a monolayer of EC was increased twofold to threefold by a defined set of C-C chemokines including RANTES, MIP1α, MIP5, and, to a lesser extent, by MIP1β and MCP-3. Most importantly, in view of the trafficking pattern of these cells, a significant proportion of DC (13% ± 4% of input cells seeded) was able to migrate across the endothelial basement membrane and, subsequently, across the endothelial barrier (reverse transmigration). The adhesion molecules and chemoattractants characterized herein are likely to underlie the complex trafficking of DC in vivo.

Original languageEnglish
Pages (from-to)207-214
Number of pages8
Issue number1
Publication statusPublished - Jul 1 1998

ASJC Scopus subject areas

  • Hematology


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