Kanabar Varsha, Tedaldi Lauren, Jiang Jingqian, Nie Xiaodan, Panina Irina, Descroix Karine, Man Francis, Pitchford Simon C, Page Clive P, Wagner Gerd K
Sackler Institute of Pulmonary Pharmacology.
Institute of Pharmaceutical Science, King's College London, Franklin Wilkins Building, London SE1 9NH, UK.
Glycobiology. 2016 Oct;26(10):1059-1071. doi: 10.1093/glycob/cww053. Epub 2016 May 27.
P-selectin glycoprotein ligand-1 (PSGL-1, CD162) is a cell-surface glycoprotein that is expressed, either constitutively or inducibly, on all myeloid and lymphoid cell lineages. PSGL-1 is implicated in cell-cell interactions between platelets, leukocytes and endothelial cells, and a key mediator of inflammatory cell recruitment and transmigration into tissues. Here, we have investigated the effects of the β-1,4-galactosyltransferase inhibitor 5-(5-formylthien-2-yl) UDP-Gal (5-FT UDP-Gal, compound 1: ) and two close derivatives on the cell surface levels of PSGL-1 on human peripheral blood mononuclear cells (hPBMCs). PSGL-1 levels were studied both under basal conditions, and upon stimulation of hPBMCs with interleukin-1β (IL-1β). Between 1 and 24 hours after IL-1β stimulation, we observed initial PSGL-1 shedding, followed by an increase in PSGL-1 levels on the cell surface, with a maximal window between IL-1β-induced and basal levels after 72 h. All three inhibitors reduce PSGL-1 levels on IL-1β-stimulated cells in a concentration-dependent manner, but show no such effect in resting cells. Compound 1: also affects the cell surface levels of adhesion molecule CD11b in IL-1β-stimulated hPBMCs, but not of glycoproteins CD14 and CCR2. This activity profile may be linked to the inhibition of global Sialyl Lewis presentation on hPBMCs by compound 1: , which we have also observed. Although this mechanistic explanation remains hypothetical at present, our results show, for the first time, that small molecules can discriminate between IL-1β-induced and basal levels of cell surface PSGL-1. These findings open new avenues for intervention with PSGL-1 presentation on the cell surface of primed hPBMCs and may have implications for anti-inflammatory drug development.
P-选择素糖蛋白配体-1(PSGL-1,CD162)是一种细胞表面糖蛋白,在所有髓系和淋巴系细胞谱系中组成性或诱导性表达。PSGL-1参与血小板、白细胞和内皮细胞之间的细胞间相互作用,是炎症细胞募集和向组织迁移的关键介质。在此,我们研究了β-1,4-半乳糖基转移酶抑制剂5-(5-甲酰噻吩-2-基)UDP-半乳糖(5-FT UDP-半乳糖,化合物1:)及其两种紧密衍生物对人外周血单个核细胞(hPBMCs)表面PSGL-1水平的影响。在基础条件下以及用白细胞介素-1β(IL-1β)刺激hPBMCs后,均对PSGL-1水平进行了研究。在IL-1β刺激后1至24小时内,我们观察到PSGL-1最初脱落,随后细胞表面PSGL-1水平升高,在72小时后达到IL-1β诱导水平与基础水平之间的最大差值。所有三种抑制剂均以浓度依赖性方式降低IL-1β刺激细胞上的PSGL-1水平,但对静息细胞无此作用。化合物1:还影响IL-1β刺激的hPBMCs中黏附分子CD11b的细胞表面水平,但不影响糖蛋白CD14和CCR2的水平。这种活性谱可能与化合物1:对hPBMCs上整体唾液酸化路易斯表位的抑制有关,我们也观察到了这一点。尽管目前这种机制解释仍属假设,但我们的结果首次表明,小分子可以区分IL-1β诱导的和基础水平的细胞表面PSGL-1。这些发现为干预致敏hPBMCs细胞表面PSGL-1的表达开辟了新途径,可能对抗炎药物的开发具有重要意义。