Puegge Johanna, Wang Yongzhi, Roller Jonas, Zhang Su, Luo Lingtao, Vollmar Brigitte, Thorlacius Henrik
Section of Surgery, Department of Clinical Sciences, Malmx00F6;, Lund University, Malmx00F6;, Sweden.
Eur Surg Res. 2016;56(1-2):19-31. doi: 10.1159/000441778. Epub 2015 Nov 18.
Extracellular histones released during cell damage have the capacity to cause tissue injury associated with increased leukocyte accumulation. However, the molecular mechanisms regulating histone-induced leukocyte recruitment remain elusive. The objective of this study was to examine the role of adhesion molecules in histone-dependent leukocyte accumulation by use of intravital microscopy of the mouse cremaster microcirculation.
Histone 3 and TNF-α were intrascrotally administered, and anti-P-selectin, anti-P-selectin glycoprotein ligand-1 (PSGL-1), anti-membrane-activated complex-1 (Mac-1), anti-lymphocyte function antigen-1 (LFA-1) antibody and neutrophil depletion antibody were injected intravenously or intraperitoneally.
Intrascrotal injection of histone 3 dose-dependently increased leukocyte recruitment. Neutrophil depletion abolished intravascular and extravascular leukocytes after histone 3 challenge, suggesting that neutrophils were the dominating leukocyte subtype responding to histone stimulation. Pretreatment with an anti-P-selectin and an anti-PSGL-1 antibody abolished histone-stimulated neutrophil rolling, adhesion and emigration. When the anti-P-selectin or the anti-PSGL-1 antibody was administrated after histone 3 stimulation, neutrophil rolling was reduced, whereas the number of firmly adherent and emigrated neutrophils were unchanged, suggesting that the inhibitory effect of blocking P-selectin and PSGL-1 on neutrophil adhesion and recruitment was due to the reduction in neutrophil rolling. Moreover, pretreatment with antibodies against Mac-1 and LFA-1 had no effect of neutrophil rolling but abolished adhesion and emigration evoked by histone 3. Thus, our data demonstrate that P-selectin and PSGL-1 play an important role in histone-induced inflammatory cell recruitment by mediating neutrophil rolling as a precondition for histone-provoked firm adhesion and emigration in vivo. Moreover, we conclude that both Mac-1 and LFA-1 are critical in supporting histone-provoked firm adhesion of neutrophils to endothelial cells.
These novel findings define specific selectins and integrins as potential targets for pharmacological intervention in histone-dependent inflammatory diseases.
细胞损伤时释放的细胞外组蛋白能够导致与白细胞聚集增加相关的组织损伤。然而，调节组蛋白诱导的白细胞募集的分子机制仍不清楚。本研究的目的是通过小鼠提睾肌微循环活体显微镜检查来研究黏附分子在组蛋白依赖性白细胞聚集中的作用。
阴囊内注射组蛋白3和肿瘤坏死因子-α，并静脉内或腹腔内注射抗P-选择素、抗P-选择素糖蛋白配体-1（PSGL-1）、抗膜活化复合物-1（Mac-1）、抗淋巴细胞功能抗原-1（LFA-1）抗体以及中性粒细胞清除抗体。
阴囊内注射组蛋白3可剂量依赖性地增加白细胞募集。组蛋白3刺激后，中性粒细胞清除可消除血管内和血管外的白细胞，这表明中性粒细胞是对组蛋白刺激作出反应的主要白细胞亚型。抗P-选择素和抗PSGL-1抗体预处理可消除组蛋白刺激的中性粒细胞滚动、黏附和移出。在组蛋白3刺激后给予抗P-选择素或抗PSGL-1抗体时，中性粒细胞滚动减少，而牢固黏附和移出的中性粒细胞数量不变，这表明阻断P-选择素和PSGL-1对中性粒细胞黏附和募集的抑制作用是由于中性粒细胞滚动减少所致。此外，抗Mac-1和抗LFA-1抗体预处理对中性粒细胞滚动没有影响，但可消除组蛋白3引起的黏附和移出。因此，我们的数据表明，P-选择素和PSGL-1通过介导中性粒细胞滚动，作为组蛋白在体内引发牢固黏附和移出的前提条件，在组蛋白诱导的炎症细胞募集中发挥重要作用。此外，我们得出结论，Mac-1和LFA-1在支持组蛋白引发的中性粒细胞与内皮细胞的牢固黏附中都至关重要。
这些新发现确定了特定的选择素和整合素是组蛋白依赖性炎症性疾病药物干预的潜在靶点。
