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VASCULAR BIOLOGY

A proteasome inhibitor reduces concurrent, sequential, and long-term IL-1- and TNF--induced ECAM expression and adhesion

Department of Chemical Engineering, Ohio University, Athens, Ohio 45701

Submitted 17 March 2003 ; accepted in final form 2 June 2003

A promising approach for reducing aberrant leukocyte-endothelial adhesion during pathological inflammation is to inhibit endothelial cell adhesion molecule (ECAM) expression at the transcription level. Several compounds have been shown to decrease cytokine-induced upregulation of ECAMs primarily by modulating the activity of transcription factors [e.g., nuclear factor-B (NF-B)]. The majority of the in vitro studies have focused on the effect of transcription inhibitors on endothelial cells exposed to a single cytokine [primarily tumor necrosis factor- (TNF-)] for a relatively short period of time (primarily 4-6 h). However, in the in vivo setting, multiple cytokines [e.g., interleukin-1 (IL-1) and TNF-] may be present for extended periods of time. Thus we studied the effects of a transcription inhibitor, the proteasome inhibitor lactacystin, on ECAM expression and myeloid (HL60) cell adhesion to human umbilical vein endothelial cells (HUVEC) activated by concurrent, sequential, and long-term (24 h) treatment with IL-1 and TNF-. We show, for the first time, that lactacystin inhibits 1) 4-h concurrent IL-1- and TNF--induced expression of E-selectin, VCAM-1, ICAM-1, and HL60 cell adhesion to HUVEC; 2) 4-h TNF--induced expression of E-selectin, VCAM-1, and HL60 cell adhesion to HUVEC that have become desensitized to IL-1 activation; 3) 24-h TNF--induced expression of E-selectin and VCAM-1 but not ICAM-1; and 4) 24-h TNF--induced HL60 cell adhesion to HUVEC. Combined, our results demonstrate that a proteasome inhibitor can reduce concurrent, sequential, and long-term IL-1- and TNF--induced ECAM expression and myeloid cell adhesion.

endothelial cell adhesion molecules; inflammation; cytokines; proteasome inhibitor

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