Platelet thrombus formation in eHUS is prevented by anti-MBL2
Platelet thrombus formation in eHUS is prevented by anti-MBL2
复制标题
DOI：
10.1371/journal.pone.0220483
复制
发表时间：
2019-12-27
期刊：
影响因子：
3.7
通讯作者：
Grabowski, E. F.
中科院分区：
文献类型：
--
作者：
Kushak, R., I;Boyle, D. C.;Grabowski, E. F.
E.由产滋贺毒素细菌引起的大肠杆菌相关溶血性尿毒综合征（流行性溶血性尿毒综合征，eHUS）的特征在于血小板减少症、微血管病性溶血性贫血和急性肾损伤，其在高达65%的受影响患者中引起急性肾衰竭。我们假设补体激活的甘露糖结合凝集素（MBL）途径在人eHUS中起重要作用，因为我们先前证明了滋贺毒素-2（Stx-2）的注射导致小鼠肾小球中的纤维蛋白沉积，其被抗MBL-2抗体3F 8的共注射阻断。然而，在受影响的小鼠肾小球血小板血栓形成的标志物没有划定。为了研究3F 8对血小板血栓形成标志物的影响，我们使用来自我们的小鼠模型（MBL-2+/-F Mbl-A/C-/-; MBL 2 KI小鼠）的肾切片。对照组的小鼠接受PBS，而第二组的小鼠接受Stx-2，第三组的小鼠接受3F 8和Stx-2。使用双重免疫荧光（IF），然后进行数字图像分析，对肾切片进行纤维蛋白（原）和CD 41（血小板标记物）、von-Willebrand因子（内皮细胞和血小板标记物）和podocin（足细胞标记物）染色。在来自具有HUS的小鼠和人的切片上进行电子显微镜（EM）。注射Stx-2导致肾小球中纤维蛋白和血小板增加，而施用3F 8与Stx-2将血小板和纤维蛋白降低至对照水平。EM研究证实IF观察到的CD 41阳性物体为血小板。通过注射Stx-2引起的血小板数量和纤维蛋白水平的增加与3F 8防止的血小板-纤维蛋白血栓的产生一致。
E. coli associated Hemolytic Uremic Syndrome (epidemic hemolytic uremic syndrome, eHUS) caused by Shiga toxin-producing bacteria is characterized by thrombocytopenia, microangiopathic hemolytic anemia, and acute kidney injury that cause acute renal failure in up to 65% of affected patients. We hypothesized that the mannose-binding lectin (MBL) pathway of complement activation plays an important role in human eHUS, as we previously demonstrated that injection of Shiga Toxin-2 (Stx-2) led to fibrin deposition in mouse glomeruli that was blocked by co-injection of the anti-MBL-2 antibody 3F8. However, the markers of platelet thrombosis in affected mouse glomeruli were not delineated. To investigate the effect of 3F8 on markers of platelet thrombosis, we used kidney sections from our mouse model (MBL-2+/-F Mbl-A/C-/-; MBL2 KI mouse). Mice in the control group received PBS, while mice in a second group received Stx-2, and those in a third group received 3F8 and Stx-2. Using double immunofluorescence (IF) followed by digital image analysis, kidney sections were stained for fibrin(ogen) and CD41 (marker for platelets), von-Willebrand factor (marker for endothelial cells and platelets), and podocin (marker for podocytes). Electron microscopy (EM) was performed on ultrathin sections from mice and human with HUS. Injection of Stx-2 resulted in an increase of both fibrin and platelets in glomeruli, while administration of 3F8 with Stx-2 reduced both platelet and fibrin to control levels. EM studies confirmed that CD41-positive objects observed by IF were platelets. The increases in platelet number and fibrin levels by injection of Stx-2 are consistent with the generation of platelet-fibrin thrombi that were prevented by 3F8.