VEGF expression in human macrophages is NF-κB-dependent: studies using adenoviruses expressing the endogenous NF-κB inhibitor IκBα and a kinase-defective form of the IκB kinase 2

• DOI: 10.1242/jcs.00286
• Publication Date: 2003-01-21T20:45:47Z
• Authors: Serafim Kiriakidis; Evangelos Andreakos; Claudia Monaco; Brian Foxwell; Marc Feldmann; Ewa Paleolog
• Keywords: Inflammation; Lipopolysaccharides; Lymphokines; 0303 health sciences; MAP Kinase Signaling System; Macrophages; CD40 Ligand; Genetic Vectors; NF-kappa B; Down-Regulation; Endothelial Growth Factors; Antibodies; Adenoviridae; I-kappa B Kinase; 03 medical and health sciences; Gene Expression Regulation; NF-KappaB Inhibitor alpha; Mutation; Humans; Intercellular Signaling Peptides and Proteins; I-kappa B Proteins; Cells, Cultured

Vascular endothelial growth factor (VEGF) is the most endothelial cell-specific angiogenic factor characterised to date, and it is produced by a variety of cell types. In macrophages, VEGF has been shown to be upregulated by the inflammatory mediator lipopolysaccharide (LPS) and by engagement of CD40 by CD40 ligand (CD40L). Because LPS and CD40L activate nuclear factor-κB (NF-κB) in monocytes, we investigated in this study whether VEGF production in macrophages, when stimulated with either LPS or CD40L, is NF-κB-dependent. We used adenoviral constructs over-expressing either IκBα (AdvIκBα), the endogenous inhibitor of NF-κB, or a kinase-defective mutant of IKK-2 (AdvIKK-2dn), an upstream activator of IκBα, to infect normal human monocyte-derived macrophages. We observed that LPS-induced production of VEGF in human macrophages was almost completely inhibited (>90%) following adenoviral transfer of IκBα. In addition, we observed significant inhibition of the CD40L-induced VEGF production in macrophages following infection with AdvIκBα. Expression of IKK-2dn in macrophages decreased VEGF production in response to LPS or CD40L by approximately 50%, suggesting that in addition to IKK-2, other kinases might be involved in NF-κB activation. These results show for the first time that VEGF production in human macrophages is NF-κB dependent. NF-κB regulates many of the genes involved in immune and inflammatory responses, and our study adds the angiogenic cytokine VEGF to the list of NF-κB-dependent cytokines.