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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, and
Paola Allavena
From the Department of Immunology and Cell Biology, "Mario
Negri" Institute, Milan; and the Section of General Pathology,
University of Brescia, Brescia, Italy.
Dendritic cells (DC) are migratory cells which exhibit complex
trafficking properties in vivo, involving interaction with vascular and
lymphatic endothelium and 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 a 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, MIP1 , MIP5, and, to a lesser extent, by MIP1
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.
Blood, Vol. 92 No. 1 (July 1), 1998:
pp. 207-214
© 1998 by The American Society of Hematology.
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