High-density lipoprotein (HDL) interacts with various cells, particularly macrophages, in functional cell-HDL interactions. Here, we found that HDL protein quality and lipid quality play critical roles in HDL functions. HDL fractions from healthy volunteers (HDL Healthy) and patients with recurrent coronary atherosclerotic disease (HDL CAD) were prepared. To analyse functional HDL-macrophage interactions, macrophages were co-incubated with each HDL, and lipid mediator production was assessed by liquid chromatography/mass spectrometry-based metabololipidomics. HDL Healthy treatment attenuated the pro-inflammatory lipid mediator production, particularly that of leukotriene (LT) B 4, and this treatment enhanced lipoxin (LX) B 4 and resolvin (Rv) E2 production. HDL Healthy treatment enhanced the proteasome-mediated degradation of the LTB 4-producing enzyme 5-lipoxygenase (LO) in activated macrophages; however, HDL CAD did not show these anti-inflammatory effects. HDL Healthy was engulfed by macrophages via clathrin-mediated endocytosis, which was a critical step in 5-LO/LTB 4 regulation. We also found that HDL CAD showed higher levels of the LTB 4-producing enzymes and thus promoted LTB 4 production from HDL CAD. In addition, LTB 4 attenuated HDL endocytosis, HDL-mediated 5-LO degradation in macrophages, and HDL-derived augmentation of macrophage phagocytosis. These results indicated that local LTB 4 produced de novo from HDL CAD regulates HDL-macrophage functional interactions and plays critical roles in dysfunctional, inflammatory HDL characteristics.