NF-κB-dependent fractalkine induction in rat aortic endothelial cells stimulated by IL-1β, TNF-α, and LPS

Abstract

Fractalkine is an endothelial cell-derived CX3C chemokine that is chemotactic mainly to mononuclear cells. Fractalkine was induced in rat aortic endothelial cells (RAEC) by interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and lipopolysaccharide (LPS) transcriptionally and translationally. This induction correlated with increased NF-kappaB DNA binding activity as determined by gel mobility shift assay. Supershift assays revealed that the NF-kappaB subunits p50 and p65 were responsible for kappaB binding. Accordingly, we examined the role of NF-kappaB in fractalkine induction in RAEC through the use of an adenovirus-mediated mutant IkappaB as a specific inhibitor. Delivery of a dominant-negative form of IkappaBalpha in RAEC dramatically reduced the induction of fractalkine by these stimuli, suggesting a role for NF-kappaB activation in fractalkine induction. The inhibition of fractalkine expression by two potent NF-kappaB inhibitors, sulfasalazine and sanguinarine, further supported the central role of NF-kappaB in fractalkine transcription regulation and suggested a novel therapeutic target aimed at modulating leukocyte endothelial cell interaction.

Suppressor of cytokine signalling (SOCS)-3 protects beta cells against IL-1?-mediated toxicity through inhibition of multiple nuclear factor-?B-regulated proapoptotic pathways

“…Among these, nine genes, namely iNOS, ICAM, complement C3, Mob-1, MIP-1, CX3C, NF-κB-p105, IRF-1 and fibrinogen-γ are usually regulated in an NF-κB-dependent manner [3,18,35,36,37,38,39,40,41], suggesting that inhibition of IL-1β signalling by SOCS-3 affects the NF-κB pathway. Furthermore, all of these transcripts were previously shown to be independent of induced NO production [28].…”

Mononuclear Cell-Infiltrate Inhibition by Blocking Macrophage-Derived Chemokine Results in Attenuation of Developing Crescentic Glomerulonephritis

“…41 Three g of total RNA from each sample were used for RNase protection assay, following a previously described protocol. 45,46 …”

15‐Deoxy‐Δ12,14‐prostaglandin J2 inhibitsCX3CL1/fractalkine expression in human endothelial cells

“…WY14643 and ciglitazone were from Calbiochem (San Diego, CA). Primer oligo(dT) [12][13][14][15][16][17][18] and MMulv reverse transcriptase were from Gibco-BRL (Gainthersburg, MD). RNeasy total RNA isolation kit and Taq DNA polymerase were purchased from Qiagen (Hilden, Germany).…”

“…Single-strand cDNA for a PCR template was synthesized from 1 µ g of total RNA using a primer oligo(dT) [12][13][14][15][16][17][18] and M-Mulv reverse transcriptase under the conditions indicated by the manufacturer. Specific primers were designed from cDNA sequences for fractalkine, ICAM-1, VCAM-1, PPAR-α , PPAR-γ and glyceraldehyde-3-phosphate dehydrogenase (GAPDH).…”

“…13 Interferon-γ is also found to stimulate endothelial fractalkine expression. 14,15 Interferon-γ stimulates fractalkine expression in other types of cells such as bronchial epithelial cells 16 and brain astrocytes. 17 Fractalkine is incorporated into the cell surface membrane of activated endothelial cells and promotes strong adhesion of T lymphocytes and monocytes.…”