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Endothelial Cells Exposed to Anoxia/Reoxygenation Are Hyperadhesive to T‐lymphocytes: Kinetics and Molecular Mechanisms
Author(s) -
KOKURA SATOSHI,
WOLF ROBERT E.,
YOSHIKAWA TOSHIKAZU,
ICHIKAWA HIROSHI,
GRANGER D. NEIL,
AW TAK YEE
Publication year - 2000
Publication title -
microcirculation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.793
H-Index - 83
eISSN - 1549-8719
pISSN - 1073-9688
DOI - 10.1111/j.1549-8719.2000.tb00739.x
Subject(s) - kinetics , chemistry , hypoxia (environmental) , microbiology and biotechnology , biophysics , immunology , biology , oxygen , organic chemistry , physics , quantum mechanics
Objective : The objectives of this study were to 1) determine the time‐course of T‐lymphocyte adhesion to monolayers of human umbilical vein endothelial cell (HUVEC) that were exposed to 60 min of anoxia followed by 24 h of reoxygenation, and 2) define the mechanisms responsible for the hyperadhesivity of post‐anoxic HUVEC to human T‐lymphocytes. Methods : Human peripheral blood mononuclear leukocytes were isolated from heparinized peripheral blood. T‐lymphocytes were obtained by negative selection using a MACS column. HUVEC monolayers were exposed to anoxia/reoxygenation (A/R), and then reacted with 51 Cr ‐labeled T‐lymphocytes in adhesion assays. Results : A/R leads to an increased adhesion of T‐lymphocytes to HUVEC monolayers, with peak responses occurring at 8 h after reoxygenation. This adhesion response was largely attributed to the CD4 + T‐cell subset. The hyperadhesivity of A/R‐exposed HUVEC was inhibited by monoclonal antibodies directed against either LFA‐1, VLA‐4, ICAM‐1, or VCAM‐1, indicating a contribution of these adhesion molecules and their ligands. Moreover, T‐cell hyperadhesivity was attenuated by anti‐ IL‐8, consistent with a role for this chemokine in the adhesion response. Protein synthesis inhibitors (actinomycin D and cycloheximide) as well as chemical inhibitors of (and binding ds ‐oligonucleotides to) NFκB and AP‐1 significantly attenuated the A/R‐induced T‐lymphocyte adhesion responses. The kinetics of VCAM‐1 on post‐anoxic HUVEC correlated with the T‐lymphocyte adhesion response. Conclusions : A/R elicits a T‐lymphocyte‐endothelial cell adhesion response that involves transcription‐dependent surface expression of VCAM‐ 1.