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

Abstract

Tumor necrosis factor (TNF) and interleukin 1 are known to initiate endothelial vascular cell adhesion molecule (VCAM)-1 transcription primarily by activating nuclear factor (NF)-κB, which translocates to the nucleus. In addition to two NF-κB elements found within the minimal cytokine-inducible VCAM-1 promoter, an interferon-related factor (IRF) element (IRF-1) has been identified close to the transcription initiation site, suggesting that cytokines that induce IRF-1 might affect VCAM-1 expression levels. We therefore investigated the effects of interferons (IFNs), which strongly induce IRF-1, on VCAM-1 transcription and expression. We show that IFN-α and -γ enhance TNF-induced VCAM-1 mRNA transcription and protein expression in human endothelial cells. IFN enhancement of TNF-induced expression is also seen using chloramphenicol acetyl transferase reporter genes linked to the minimal cytokine inducible VCAM-1 promoter. Nuclear IRF-1 is the molecular basis of IFN enhancement, because (a) IFN plus TNF–treated cells displayed increased nuclear IRF-1 levels and increased IRF-1 binding to the VCAM-1 promoter, compared with cells treated with TNF alone; (b) kinetics of nuclear IRF-1 levels correlated with VCAM-1 mRNA levels; (c) transfection with an IRF-1 construct substituted for IFN treatment; and (d) transfection with an expression construct encoding IRF-2, a competitive inhibitor of IRF-1, reduced TNF-induced VCAM-1 expression. Our experiments show that IFN amplifies TNF-induced VCAM-1 expression at the transcriptional level by an IRF-1–dependent pathway.

Induction of Endothelial-Leukocyte Interaction by Interferon-γ Requires Coactivation of Nuclear Factor-κB

“…5 During the preparation of the present article, a similar synergistic interaction has been reported for IFN-␥ and TNF. 7 However, the present study demonstrates that low concentrations of IL-1 are unable to induce VCAM-1 gene transcription without the added effects of IFN-␥, which alone also cannot induce VCAM-1 expression.…”

“…Nuclei from 3 to 5ϫ10 7 endothelial cells were prepared 16 hours after stimulation with IL-1␣ (low and high concentrations) and IFN-␥, either alone or in combination, and in vitro transcription with [␣-32 P]UTP (800 Ci/mmol) was performed as described. 19 Linearized plasmids (1 g) were immobilized on nylon membranes, hybridized to radiolabeled transcripts (Ϸ5 to 8ϫ10 7 cpm/mL) at 45°C for 48 hours in hybridization buffer containing 50% formamide, 5ϫ SSC, 2.5ϫ Denhardt's solution, 25 mmol/L sodium phosphate buffer (pH 6.5), 0.1% SDS, and 250 g/mL salmon sperm DNA, and washed in 1ϫ SSC/0.1% SDS at 65°C before autoradiography.…”

“…5 Indeed, IRF-1 has been found to synergize with NF-B in transactivating cytokine-inducible genes, such as inducible type II NO synthase 6 and VCAM-1. 7 A major product of activated T lymphocytes, interferon (IFN)-␥, induces functional changes in the vascular endothelium, including the expression of major histocompatibility complex (MHC) class II antigens. [8][9][10] Activated T lymphocytes and MHC-II-positive vascular smooth muscle cells (SMCs) are colocalized within atherosclerotic lesions, 11,12 and recent evidence suggests that T lymphocytes contribute to the development and rupture of atherosclerotic plaques.…”

Dermal Microvascular Endothelial Cells Express the 180-kDa Macrophage Mannose Receptor In Situ and In Vitro

“…Total RNA was isolated and reverse transcribed as previously described (29). Primer sequences for the MR amplify cDNA only and were described by Lu et al (30).…”

Alpha-V-beta-3• integrin-mediated adenoviral transfer of interleukin-12 at the periphery of hepatic colon cancer metastases induces VCAM-1 expression and T-cell recruitment

“…The synergy of adoptive T-cell therapy and IL-12 gene transfer at the effector phase could be explained if some-how IL-12 could condition the liver metastasis for lymphocyte infiltration. It has been recently reported that VCAM-1 (CD106) and ICAM-1 (CD54) are upregulated in the vessels of murine ovarian cancers when recombinant IL-12 is given systemically and its expression is induced by cytokines such as TNF␣, IFN␥, and IL-1 (27)(28)(29). VCAM-1 is an endothelial counterreceptor for the VLA-4 integrin (30), which is expressed in a molecular conformation with high avidity for VCAM-1 on activated T-cells (31) and which mediates lymphocyte diapedesis (32).…”