Comprehensive regulatory system of PAMPs/DAMPs by TM-PC/EPCR system

内皮血栓调节蛋白(TM)结合DAMPS(损伤相关分子模式)；HMGB1是DAMPS的代表，组蛋白。此外，TM还与PAMPs(病原体相关分子模式)结合，将PAMPs的代表脂多糖(LPS)结合到分子的凝集素样域上，并中和它们的细胞病变活性，包括促炎和促凝血活性。HMGB1与TM结合，被凝血酶/TM复合体降解，生成N端缺失的HMGB1，命名为DES-HMGB1。DES-HMGB1与受体RAGE和TLR2-2、-4结合。然而，与完整的HMGB1相比，结合亲和力较弱，提示该分子的DES形式可能在HMGB1及其受体之间形成了一个负反馈回路，调节HMGB1的不同活性。因此，检测DES-HMGB1可能对弥漫性血管内凝血(DIC)、脓毒症、全身炎症反应综合征(SIRS)等疾病状态提供有意义的信息。因此，我们尝试建立DES-HMGB1的特异性检测方法。我们得到了与DES-HMGB1反应的特异性单抗。用该单抗建立了特异性的酶联免疫吸附试验(ELISA)。该方法成功地检测到DES-HMGB1，完整形式的污染率为10%。DES-HMGB1可增加DIC患者血清中DES-HMGB1的表达，重组TM对DES-HMGB1的作用尤为明显。然而，增加的DES-HMGB1迅速下降，表明DES形式迅速翻转。此外，一些患者即使接受重组TM治疗，DES-HMGB1也没有增加，提示存在或无应答病例。因此，DES-HMGB1的检测有望为DIC、全身炎症反应综合征、脓毒症等疾病的治疗提供新的信息。

Endothelial thrombomodulin(TM) bounds DAMPs(Damage Associated Molecular Patterns);HMGB1,a representative DAMPs, histones. Moreover TM also bounds PAMPs(Pathogen Associated Molecular Patterns) ;lipopolysaccaride(LPS), a representative PAMPs, onto the lectin-like domain of the molecule and neutralizes their cytopathic activities, including proinflammatory and procoagulant activities. HMGB1 bound to TM, degraded by thrombin/TM complex and generated the N-terminus deleted HMGB1, named as des-HMGB1. Des-HMGB1 bound onto the receptors RAGE and TLR2-2, -4. However the binding affinity was weak compared to intact HMGB1, suggesting des-form of the molecule may configure a negative feedback loop among HMGB1 and its receptors regulating the diverse activities of HMGB1. Based on the findings, assay of des-HMGB1.may provide significant information in such morbid states including Disseminated Intravascular Coagulation(DIC), sepsis, systemic inflammatory response sybdrome(SIRS) and so on. Therefore we tried to establish specific assay method of des-HMGB1. We got specific monoclonal antibodies to react with the des-HMGB1. Using the monoclonal antibody, we established specific enzyme linked immune sorbent assay(ELISA). The ELISA successfully detected des-HMGB1 with 10% contamination of intact form. Des-HMGB1 was increased the serum from DIC, especially treated with recombinant TM. However the increased des-HMGB1 was decreased rapidly suggesting the rapid turn-over of the des-form. Moreover some patients showed no increased in des-HMGB1 even with recombinant TM treatment, suggesting of the presence or non-responder cases. Thus assessment of des-HMGB1 may be expected to provide a novel information in the DIC, SIRS, sepsis and so on with or without treatment.