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-CD47 Interactions Are Required for the Transmigration of Monocytes Across Cerebral Endothelium1
Departments of
* Molecular Cell Biology and
Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands
Monocyte infiltration into inflamed tissue requires their initial
arrest onto the endothelial cells (ECs), followed by firm adhesion and
subsequent transmigration. Although several pairs of adhesion molecules
have been shown to play a role in the initial adhesion of monocytes to
ECs, the mechanism of transendothelial migration is poorly defined. In
this study, we have investigated the role of signal-regulatory protein
(SIRP)
-CD47 interactions in monocyte transmigration across brain
ECs. CD47 expression was observed in vivo on cerebral endothelium of
both control animals and animals suffering from experimental allergic
encephalomyelitis. To investigate whether SIRP-CD47 interactions are
instrumental in the trafficking of monocytes across cerebral EC
monolayers, in vitro assays were conducted in which the migration of
monocytes, but not adhesion, was found to be effectively diminished by
blocking SIRP and CD47 on monocytes and ECs, respectively. In this
process, SIRP was found to interact solely with its counterligand
CD47 on ECs. Overexpression of the CD47 molecule on brain ECs
significantly enhanced monocytic transmigration, but did not affect
adhesion. SIRP-CD47-mediated transendothelial migration involved Gi
protein activity, a known signaling component of CD47. Finally,
cross-linking of CD47 on brain ECs induced cytoskeletal reorganization
of the endothelium, a process that was Gi protein independent. These
data provide the first evidence that the interaction of CD47 with its
monocytic counterligand SIRP is of importance in the final step of
monocyte trafficking into the brain, a critical event in the
development of neuroinflammatory diseases.
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