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Matthew D. Silverman, David O. Zamora, Yuzhen Pan, Paul V. Texeira, Stephen R. Planck, James T. Rosenbaum; Cell Adhesion Molecule Expression in Cultured Human Iris Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2001;42(12):2861-2866.
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© ARVO (1962-2015); The Authors (2016-present)
purpose. To develop a method to isolate human iris microvascular endothelial
cells (HIECs) for exploring their constitutive and inflammatory
agent-modulated expression of intercellular adhesion molecules (ICAM)-1
and -2, vascular cell adhesion molecule (VCAM)-1, and E-selectin.
methods. Endothelial cells from collagenase-digested irises were isolated on the
basis of their expression of platelet endothelial cell adhesion
molecule (PECAM)-1, using antibody-coupled magnetic beads. Cells were
characterized as endothelial based on morphologic criteria, their
expression of PECAM-1 and von Willebrand factor, their uptake of
acetylated low-density lipoprotein, and their ability to form
capillary-like networks on a synthetic basement membrane. Constitutive
and inflammatory agent–modulated expression of ICAM-1 and -2, VCAM-1,
and E-selectin was evaluated by the reverse transcription–polymerase
chain reaction, enzyme-linked immunocellular assays (ELICAs), Western
blot analysis, and functional studies of leukocyte adhesion to HIEC
results. HIECs constitutively expressed mRNA and protein for ICAM-1 and -2, but
only low to nondetectable levels of VCAM-1 or E-selectin. When
stimulated with endotoxin- or tumor necrosis factor (TNF)-α, ICAM-1,
VCAM-1, and E-selectin were potently and time- and dose-dependently
upregulated at both the message and protein levels. By contrast, ICAM-2
message and protein were slowly downregulated by inflammatory agents
over time, but nonetheless remained present and functional. Overall,
cytokine- or endotoxin-activation of HIECs resulted in enhanced
adhesiveness for leukocytes.
conclusions. ICAM-1, VCAM-1, and E-selectin have been previously implicated in
mediating anterior ocular inflammation. This is a report of the
selective isolation of HIECs, with a demonstration of differential
expression and regulation of these adhesion molecules in them. In
addition, this is the first demonstration of the regulated expression
of ICAM-2 in any ocular microvascular cells.
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