Downregulation by tumor necrosis factor-alpha of monocyte CCR2 expression and monocyte chemotactic protein-1-induced transendothelial migration is antagonized by oxidized low-density lipoprotein: a potential mechanism of monocyte retention in atherosclerotic lesions.

The subintimal infiltration with monocytes is crucially involved in the development of complex atherosclerotic plaques. Monocyte chemotactic protein-1 (MCP-1) and its receptor CCR2 are important for monocyte extravasation and formation of atherosclerotic lesions. However, mechanisms of monocyte persistence in atherosclerotic plaques remain to be elucidated. Flow cytometric analysis revealed that monocytoid Mono Mac 6 cells that had transmigrated endothelium towards a MCP-1 gradient expressed higher levels of CCR2 than the non-migratory fraction, while input cells were intermediate, suggesting that high CCR2 levels are essential for transendothelial chemotaxis. Pretreatment of Mono Mac 6 cells or isolated human blood monocytes with the inflammatory cytokine tumor necrosis factor (TNF)-alpha dose- and time-dependently reduced MCP-1-induced transendothelial chemotaxis, which was inhibited by the CCR2 receptor antagonist 9-76 analog. This was paralleled by a decrease in CCR2 surface protein and mRNA expression. as assessed by flow cytometry and reverse transcription-polymerase chain reaction, inferring that inhibition of monocyte transmigration was due to downregulation of CCR2 to levels insufficient for chemotaxis. In contrast, treatment of monocytes with oxidized low-density protein (oxLDL) containing oxidized lipids, such as cholesteryl linoleate 13-hydroxide. but not with LDL, increased CCR2 protein and mRNA expression. Notably, oxLDL counteracted the TNF-alpha-mediated downregulation of CCR2 and CCR2-dependent transendothelial chemotaxis. Macrophage-colony-stimulating factor hardly affected CCR2 expression and function, suggesting that differentiation was not responsible for effects on CCR2. In conclusion, TNF-alpha impairs MCP-1-induced transendothelial migration of monocytes by downregulating CCR2 which appears critical for migration. Exposure to oxLDL antagonized the effects of TNF-alpha, and may thus contribute to monocyte retention and perpetuation of a chronic inflammatory reaction in unstable atherosclerotic lesions.