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J. Cell Biol.,
Volume 145, Number 2, April 19, 1999 403-412
* Blood-Brain Barrier Laboratory, Department of Pharmacology, University of Connecticut Health Center, Farmington,
Connecticut 06030; and The chemokines monocyte chemoattractant
protein-1 (MCP-1) and macrophage inflammatory protein-1 Department of Neurosurgery, Yale University School of Medicine, New Haven, Connecticut 06510
(MIP-1) aid in directing leukocytes to specific
locales within the brain and spinal cord during central
nervous system inflammation. However, it remains unclear how these chemokines exert their actions across a
vascular barrier, raising speculation that interaction
with endothelial cells might be required. Therefore, experiments were performed to determine whether binding domains for these chemokines exist along the outer
surface of brain microvessels, a feature that could potentially relay chemokine signals from brain to blood. Using a biotinylated chemokine binding assay with confocal microscopy and three-dimensional image reconstruction, spatially resolved binding sites for MCP-1
and MIP- around human brain microvessels were revealed for the first time. Binding of labeled MCP-1 and MIP-1 could be inhibited by unlabeled homologous
but not heterologous chemokine, and was independent
of the presence of heparan sulfate, laminin, or collagen
in the subendothelial matrix. This is the first evidence of
specific and separate binding domains for MCP-1 and
MIP-1 on the parenchymal surface of microvessels,
and highlights the prospect that specific interactions of
chemokines with microvascular elements influence the
extent and course of central nervous system inflammation.
;
binding;
microvessels
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