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Vol. 58, Issue 1, 237-245, July 2000
Receptor-1 Coupled to
Tumor Necrosis Factor Receptor-Associated Factor 2 Stimulates
Intercellular Adhesion Molecule-1 Expression by Modulating a
Thapsigargin-Sensitive Pathway in Human Tracheal Smooth Muscle Cells
Pulmonary, Allergy and Critical Care Division, Department of
Medicine, University of Pennsylvania Medical Center, Philadelphia,
Pennsylvania
Tumor necrosis factor-
(TNF) stimulates the expression of
intercellular adhesion molecule-1 (ICAM-1) by activating the
transcription factor nuclear factor-
B (NF-B) in human airway
smooth muscle (ASM) cells. This study characterizes the receptor
involved as well as critical downstream signaling events mediating
cytokine-induced NF-B activation and ICAM-1 expression. TNF
stimulation for 1 to 4 h induced ICAM-1 expression in human ASM
cells. This rapid TNF-induced ICAM-1 expression enhanced
T-lymphocyte adhesion to ASM cells, which was inhibited by anti-ICAM-1
antibodies. Using immunostaining, we demonstrated that TNF receptors
TNFR1 and TNFR2 are expressed on native human tracheal smooth muscle.
Treatment of cells with htr-9, an antibody that specifically activates
TNFR1, also stimulated expression of ICAM-1 mRNA and protein. Utr-1, a
blocking antibody to TNFR2, did not affect TNF-mediated ICAM-1 expression. Both TNF and htr-9 increased luciferase activity in ASM
cells transfected with a NF-B reporter plasmid. Overexpression of a
dominant negative TNF receptor-associated factor 2 construct, lacking the NH2-terminal RING finger, completely abrogated
both TNF- and htr-9-mediated increases in NF-B reporter activity. Thapsigargin, an agent that depletes intracellular calcium stores, abrogated both cytokine-mediated NF-B-dependent ICAM-1 mRNA
transcription and protein expression but had no effect on IB
degradation. In addition, chelating cytosolic calcium
with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid
acetoxymethyl ester also inhibited cytokine TNF-induced ICAM-1
expression. These data suggest that TNFR1, through a TNF receptor-associated factor 2-NF-B signaling pathway, mediates TNF-induced expression of ICAM-1 on ASM cells by involving a thapsigargin-sensitive signaling pathway.
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