Institute of General Pharmacology and Toxicology, pharmazentrum frankfurt/ZAFES, Hospital of the Goethe University, Frankfurt, Germany.
Institute of Clinical Pharmacology, pharmazentrum frankfurt, Hospital of the Goethe University, Frankfurt, Germany.
Mediators Inflamm. 2017;2017:5187368. doi: 10.1155/2017/5187368. Epub 2017 Dec 11.
A balanced sphingolipid rheostat is indispensable for dendritic cell function and survival and thus initiation of an immune response. Sphingolipid levels are dynamically maintained by the action of sphingolipid enzymes of which sphingosine kinases, S1P phosphatases (SGPP-1/2) and S1P lyase (SGPL-1), are pivotal in the balance of S1P and sphingosine levels. In this study, we present that SGPP-1 and SGPL-1 are regulated in inflammatory dendritic cells and contribute to S1P fate. TLR-dependent activation caused SGPL-1 protein downregulation with subsequent decrease of enzymatic activity by two-thirds. In parallel, confocal fluorescence microscopy revealed that endogenous SGPP-1 was expressed in nuclei of naive dendritic cells and was translocated into the cytoplasmatic compartment upon inflammatory stimulation resulting in dephosphorylation of S1P. Mass spectrometric determination showed that a part of the resulting sphingosine was released from the cell, increasing extracellular levels. Another route of diminishing intracellular S1P was possibly taken by its export via ATP-binding cassette transporter C1 which was upregulated in array analysis, while the S1P transporter, spinster homolog 2, was not relevant in dendritic cells. These investigations newly describe the sequential expression and localization of the endogenous S1P regulators SGPP-1 and SGPL-1 and highlight their contribution to the sphingolipid rheostat in inflammation.
平衡的神经酰胺变阻器对于树突状细胞的功能和存活以及免疫反应的启动是必不可少的。神经酰胺水平通过神经酰胺酶的作用动态维持,其中鞘氨醇激酶、S1P 磷酸酶 (SGPP-1/2) 和 S1P 裂解酶 (SGPL-1) 在 S1P 和鞘氨醇水平的平衡中起着关键作用。在这项研究中,我们提出 SGPP-1 和 SGPL-1 在炎症性树突状细胞中受到调节,并有助于 S1P 的命运。TLR 依赖性激活导致 SGPL-1 蛋白下调,随后酶活性降低三分之二。同时,共聚焦荧光显微镜显示内源性 SGPP-1 在幼稚树突状细胞的核中表达,并在炎症刺激下转移到细胞质区室,导致 S1P 去磷酸化。质谱测定表明,一部分产生的鞘氨醇从细胞中释放出来,增加了细胞外水平。另一种减少细胞内 S1P 的途径可能是通过其通过 ATP 结合盒转运蛋白 C1 输出,该蛋白在阵列分析中上调,而 S1P 转运蛋白 spinster 同源物 2 在树突状细胞中不相关。这些研究新描述了内源性 S1P 调节剂 SGPP-1 和 SGPL-1 的顺序表达和定位,并强调了它们在炎症中对神经酰胺变阻器的贡献。