Receptors involved in the oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero3-phosphorylcholine-mediated synthesis of interleukin-8 -: A role for toll-like receptor 4 and a glycosylphosphatidylinositol-anchored protein
Receptors involved in the oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero3-phosphorylcholine-mediated synthesis of interleukin-8 -: A role for toll-like receptor 4 and a glycosylphosphatidylinositol-anchored protein
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DOI：
10.1074/jbc.m300738200
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发表时间：
2003-08-08
影响因子：
4.8
通讯作者：
Berliner, JA
中科院分区：
文献类型：
--
作者：
Walton, KA;Hsieh, X;Berliner, JA
我们先前证明，氧化的1-棕榈酰-2-花生四烯酸-sn-甘油-3-磷酸胆碱（ox-PAPC），特别是组分脂质1-棕榈酰-2-（5，6-环氧异前列烷E-2）-sn-甘油-3-磷酸胆碱增加主动脉内皮细胞中白细胞介素-8（IL-8）的合成。目前研究的目的是表征介导IL-8转录增加的受体复合物。我们表明，清道夫受体A类，I型和II型，凝集素样ox-LDL受体-1，巨噬细胞受体与胶原结构，和CD 36是不负责的IL-8的增加。使用显性负结构和反义寡核苷酸，我们证明了Toll样受体4（TLR 4）作为ox-PAPC受体介导IL-8转录的作用。我们证明，糖基磷脂酰肌醇（GPI）锚定蛋白也是必要的，因为磷脂酰肌醇特异性磷脂酶C预处理抑制ox-PAPC的效果。CD 14是一种GPI锚定蛋白，与TLR 4一起介导脂多糖的作用，但似乎不介导ox-PAPC的作用，因为ox-PAPC诱导的IL-8转录既不被抗CD 14中和抗体阻断，也不被可溶性CD 14的加入或膜CD 14的过表达所增强。相反，抗TLR 4抗体免疫沉淀37 kDa的蛋白质，也结合ox-PAPC。在用于检测GPI锚定蛋白的气溶素覆盖物中发现了相同大小的蛋白。因此，这些研究表明，ox-PAPC可能最初结合到一个37 kDa的GPI锚定蛋白，与TLR 4相互作用，诱导IL-8的转录。
We demonstrated previously that oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (ox-PAPC) and, specifically, the component lipid 1-palmitoyl-2-(5,6-epoxyisoprostane E-2)-sn-glycero-3-phosphorylcholine increase interleukin-8 (IL-8) synthesis in aortic endothelial cells. The goal of the current studies was to characterize the receptor complex mediating the increased transcription of IL-8. We demonstrate that scavenger receptor class A, types I and II, lectin-like ox-LDL receptor-1, macrophage receptor with collagenous structure, and CD36 are not responsible for the increase in IL-8. Using dominant-negative constructs and antisense oligonucleotides, we demonstrate a role for Toll-like receptor 4 (TLR4) as the ox-PAPC receptor mediating IL-8 transcription. We demonstrate that a glycosylphosphatidylinositol (GPI)-anchored protein is also necessary because phosphatidylinositol-specific phospholipase C pretreatment inhibited the effect of ox-PAPC. CD14, a GPI-anchored protein that associates with TLR4 in mediating lipopolysaccharide action, did not appear to mediate ox-PAPC action because ox-PAPC-induced IL-8 transcription was not blocked by anti-CD14 neutralizing antibodies nor was it augmented by the addition of soluble CD14 or overexpression of membrane CD14. Instead, anti-TLR4 antibodies immunoprecipitated a 37-kDa protein that also bound ox-PAPC. A protein of this same size was found in aerolysin overlays used to detect GPI-anchored proteins. Therefore, these studies suggest that ox-PAPC may initially bind to a 37-kDa GPI-anchored protein, which interacts with TLR4 to induce IL-8 transcription.