Differential regulation of vascular cell adhesion molecule-1 gene transcription by tumor necrosis factor alpha and interleukin-1 alpha in dermal microvascular endothelial cells

Abstract

As part of the inflammatory response, the localization of leukocytes depends to an important degree on cytokine-induced expression of vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells (EC). We have previously shown that VCAM-1 expression is induced on human umbilical vein EC (HUVEC) by both tumor necrosis factor alpha (TNF- alpha) and interleukin-1 alpha (IL-1 alpha), whereas on human dermal microvascular EC (HDMEC) only TNF alpha results in VCAM-1 expression. To explore molecular mechanisms responsible for these contrasting patterns of VCAM-1 induction in HUVEC versus HDMEC, we performed transcriptional activation studies with VCAM-1-based reporter constructs and in vitro binding assays using two adjacent NF-kappa B binding sequences of the VCAM-1 promoter as a DNA probe. Previous studies have established that these NF-kappa B motifs are required for cytokine-induced VCAM-1 transcription, and may further mediate cell- specific VCAM-1 gene activation by cytokines. The findings reported here demonstrate a significant HDMEC-specific attenuation of VCAM-1 gene transcription in response to IL-1 alpha, but not TNF alpha. An upstream VCAM-1 gene regulatory region distinct from the NF-kappa B sites appears to function as an IL-1 alpha-mediated transcriptional repressor within HDMEC. This repressor region conveys IL-1 alpha- dependent, but not TNF alpha-dependent, inhibition of transcription driven by a heterologous cytokine response element and promoter.

Human Microvascular Endothelial Cell Activation by IL-1 and TNF-α Stimulates the Adhesion and Transendothelial Migration of Circulating Human CD14+ Monocytes That Develop With RANKL Into Functional Osteoclasts

“…For example, VCAM‐1 has been reported by some to play a major role, ( 56 , 62 , 63 ) but by others to have little influence, ( 64 ) in monocyte adhesion to and TEM through activated endothelium. The apparent lack of a role for VCAM‐1 in our work also might reflect the use of activated HMVECs instead of HUVECs (which express relatively more VCAM‐1 than HMVECs), ( 50 , 51 , 65 ) the ability of IL‐1 to attenuate TNF‐α‐stimulated VCAM‐1 in endothelial cells, ( 66 ) potential interference in VLA‐4 binding to VCAM‐1 because of ICAM‐1 interactions with monocyte LFA‐1, ( 67 ) a more important role for VCAM‐1/VLA‐4 interactions in monocyte lateral migration than diapedesis, ( 68 ) or other reasons. PECAM‐1, which is expressed on hematopoietic and endothelial cells, promotes the postadhesion TEM of leukocytes through endothelium.…”

Transforming growth factor-alpha -induced transcriptional activation of the vascular permeability factor (VPF/VEGF) gene requires AP-2-dependent DNA binding and transactivation

“…A431 cells, HeLa cells (American Type Culture Collection, Rockville, MD) and HaCaT cells (generously provided by Dr Norbert E.Fusenig; Boukamp et al ., 1988) were cultured at 37°C and 5% CO 2 in Dulbecco's modified essential medium (Gibco Life Technologies Inc., Grand Island, NY), supplemented with 5% fetal bovine serum (Hyclone, Logan, UT), 2 mM glutamine (Gibco), 100 U/ml penicillin (Gibco), 100 μg/ml streptomycin (Gibco) and 250 μg/ml amphotericin B (Gibco). Human dermal microvascular endothelial cells (HDMEC) were isolated and cultured as previously described (Gille et al ., 1996).…”

Interferon Enhances Tumor Necrosis Factor–induced Vascular Cell Adhesion Molecule 1 (CD106) Expression in Human Endothelial Cells by an Interferon-related Factor 1–dependent Pathway

“…Nuclear protein extracts were prepared as previously described ( 21 ), normalized according their protein concentrations, and electrophoresed on SDS/8% polyacrylamide gels and blotted onto nitrocellulose in Tris buffer (25 mM) containing 192 mM glycine and 5% methanol at 100 V/15 mA for 1 h. Membranes were incubated with first-step antibodies (rabbit anti–IRF-1, 1:100; rabbit anti–NF-κB/p65, 1:2,000; rabbit anti– Sp-1, 1:500 (all Santa Cruz Biotechnology , Santa Cruz, CA) or control rabbit serum diluted in PBS containing 1% low-fat milk for 2 h at room temperature. After rinsing, blots were incubated with horseradish peroxidase–conjugated goat anti–rabbit secondary antibody (1:100,000; Bio-Rad Laboratories, Hercules, CA), visualized by chemiluminescence (ECL system; Amersham Corp. , Buckinghamshire, UK), and recorded on film.…”