Divergent effects of Tlr9 deletion in experimental late venous thrombosis resolution and vein wall injury.
Divergent effects of Tlr9 deletion in experimental late venous thrombosis resolution and vein wall injury.
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DOI：
10.1160/th14-12-1031
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发表时间：
2015-11
影响因子：
6.7
通讯作者：
Henke PK
中科院分区：
文献类型：
--
作者：
Dewyer NA;El-Sayed OM;Luke CE;Elfline M;Kittan N;Allen R;Laser A;Oostra C;Comerota A;Hogaboam C;Kunkel SL;Henke PK
深静脉血栓（DVT）通过无菌炎症反应消退。定义 DVT 的炎症反应可能会带来不涉及抗凝的新疗法。此前，我们已经证明 Toll 样受体 9 (Tlr9) 基因缺失的小鼠静脉血栓 (VT) 消退受损。在这里，我们进一步描述了 Tlr9 信号传导和无菌炎症在慢性 VT 和静脉壁反应中的作用。首先，我们发现人类存在Tlr9+细胞存在于慢性血栓形成后管腔组织中的先例。其次，在停滞性 VT 小鼠模型中，与 WT 相比，Tlr9−/− 血栓中尿酸、HMGB-1 和中性粒细胞胞外陷阱标记的瓜氨酸组蛋白 3（和细胞外 DNA）的内源性危险信号介质比 WT 更大，这与 8 和 21 天时更大的 VT 相对应。第 8 天时，Tlr9−/− 血栓中存在的 M1 型 (CCR2+) 单核细胞/巨噬细胞 (MØ) 少于 WT 对照，表明炎症细胞流入受损。使用骨髓源性单核细胞 (BMMØ) 细胞培养物，我们发现暴露于几种内源性危险信号时纤溶基因表达降低。接下来，将培养的 Tlr9+/+ BMMØ 过继转移至 Tlr9−/− 小鼠，使 8 天时的 VT 分辨率正常化。最后，尽管 Tlr9−/− 小鼠的 VT 大小在 21 天时较大，并且与内皮抗原标记物减少相关，但未发现纤维化方面的差异。这些数据表明，MØ 中的 Tlr9 信号传导对于后期 VT 缓解至关重要，与坏死清除相关，但不影响后期静脉壁纤维化。这些发现提供了对该疾病过程中无菌炎症 Tlr9 MØ 机制的深入了解。
Deep vein thrombosis (DVT) resolves via a sterile inflammatory response. Defining the inflammatory response of DVT may allow for new therapies that do not involve anticoagulation. Previously, we have shown that Toll-like receptor 9 (Tlr9) gene deleted mice had impaired venous thrombosis (VT) resolution. Here, we further characterize the role of Tlr9 signaling and sterile inflammation in chronic VT and vein wall responses. First, we found a human precedent exists with Tlr9+ cells present in chronic post thrombotic intraluminal tissue. Second, in a stasis VT mouse model, endogenous danger signal mediators of uric acid, HMGB-1, and neutrophil extracellular traps marker of citrullinated histone-3 (and extracellular DNA) were greater in Tlr9−/− thrombi as compared with WT, corresponding with larger VT at 8 and 21d. Fewer M1 type (CCR2+) monocyte/macrophages (MØ) were present in Tlr9−/− thrombi than WT controls at 8d, suggesting an impaired inflammatory cell influx. Using bone marrow derived monocyte (BMMØ) cell culture, we found decreased fibrinolytic gene expression with exposure to several endogenous danger signals. Next, adoptive transfer of cultured Tlr9+/+ BMMØ to Tlr9−/− mice normalized VT resolution at 8d. Lastly, although the VT size was larger at 21d in Tlr9−/− mice and correlated with decreased endothelial antigen markers, no difference in fibrosis was found. These data suggest that Tlr9 signaling in MØ is critical for later VT resolution, is associated with necrosis clearance, but does not affect later vein wall fibrosis. These findings provide insight into the Tlr9 MØ mechanisms of sterile inflammation in this disease process.