Reactive oxygen species (ROS), as well reactive nitrogen species (RNS), are products of normal cellular metabolism; however, cells of the immune system produce both superoxide anion (O2 ?) and nitric oxide (NO) during the oxidative burst brought on during inflammatory processes [18]. In contrast, the lipid moiety of oxLDL-IC was detected in the endosomal compartment, whereas its apolipoprotein moiety advanced to the lysosomal compartment. Cells treated with oxLDL-IC prior to oxLDL exhibited co-localization of internalized lipid moieties from both oxLDL and oxLDL-IC in the endosomal compartment. This sequential treatment likely inhibited oxLDL lipid moieties from trafficking to the lysosomal compartment. In RAW 264.7 macrophages, oxLDL-IC but not oxLDL induced GFP-tagged heat shock protein 70 (HSP70) and HSP70B’, which co-localized with the lipid moiety of oxLDL-IC in the endosomal compartment. This suggests that HSP70 family members might prevent the degradation of the internalized lipid moiety of oxLDL-IC by delaying its advancement to the Amyloid b-peptide (42-1) (human) lysosome. The data also showed that mitochondrial membrane potential was decreased and generation of reactive oxygen and nitrogen species was increased in U937 cell treated with oxLDL compared to Rabbit Polyclonal to GLB1 oxLDL-IC. Conclusions/Significance Findings suggest that lipid and apolipoprotein moieties of oxLDL-IC traffic to separate cellular compartments, and that HSP70/70B’ might sequester the lipid moiety of oxLDL-IC in the endosomal compartment. This mechanism could ultimately influence macrophage function and survival. Furthermore, oxLDL-IC might regulate the intracellular trafficking of free oxLDL possibly through the induction of HSP70/70B’. Introduction An early event in atherosclerosis is the engorgement of macrophages with lipids. It is well established that activated macrophages become lipid-laden foam cells by taking up oxidatively modified low-density lipoprotein (oxLDL), leading to the accumulation of cholesteryl esters (CE) [1]. Circulating oxLDL elicits the production of auto-immune antibodies, predominantly of the pro-inflammatory IgG1 and IgG3 isotypes, resulting in the formation of oxLDL-containing immune complexes (oxLDL-IC) [2], [3], [4]. While both oxLDL and oxLDL-IC have been detected in human atherosclerotic plaques [5], oxLDL-IC are considerably more efficient than oxLDL in the induction of foam cell formation [6]. We and others have shown that human monocytic cells exposed to oxLDL have reduced cell survival compared to those exposed to oxLDL-IC [7], [8]. Furthermore macrophages exposed to oxLDL-IC result in the release of the pro-inflammatory and plaque destabilizing factors that promote lesion progression [9], [10], [11]. The internalization of lipids in macrophages occurs through mechanisms involving different cell surface receptors. The macrophage scavenger receptors are a family of proteins which include scavenger receptors class A (macrophage scavenger receptor I and II, MSR-I and MSR-II), and class B (SR-BI and CD36). Macrophage scavenger receptors from both classes bind modified LDL [12], [13], and mediate its delivery to lysosomes for processing and degradation [14]. In contrast, oxLDL-IC are predominantly internalized through the FC receptor I (FC RI) [15]. However, the temporal and spatial intracellular localization of lipid and apolipoprotein moieties of oxLDL-IC, and how trafficking of these moieties influences the formation, activation and survival of foam cells are still obscure. In a recent study we showed that in macrophages, internalized oxLDL-IC induces a member of the HSP70 family, heat shock protein 70B’ (HSP70B’), which co-localizes with the lipid moiety of oxLDL-IC [16]. In the current study we investigated the effect of HSP70 and HSP70B’ around the advancement of internalized moieties of oxLDL-IC to the lysosomal compartment. Based on experimental evidence and clinical studies, oxidative and nitrosative stresses have been shown to be induced by atherosclerosis risk factors and to contribute to the onset and development of atherosclerotic vascular damage [17]. Reactive oxygen species (ROS), as well reactive nitrogen species (RNS), are products of normal cellular metabolism; however, cells of the immune system produce both superoxide anion (O2 ?) and nitric oxide (NO) during the oxidative burst brought on during inflammatory processes [18]. The dynamic interactions between endogenous ROS/RNS and intracellular signaling pathways may play a key role in the activation of Amyloid b-peptide (42-1) (human) macrophages. It has been found that the generation of ROS and RNS does not completely deplete intracellular antioxidants, rather regulates the atherogenic process by modulating intracellular signaling pathways affecting inflammatory cell adhesion, migration, proliferation, and differentiation [19]. Nonetheless, overproduction of ROS/RNS or a deficiency of enzymatic or non-enzymatic antioxidants may cause biological damage to cellular lipids, Amyloid b-peptide (42-1) (human) protein, and DNA resulting in cell loss Amyloid b-peptide (42-1) (human) of life [20]. Mitochondria Amyloid b-peptide (42-1) (human) are both main.