Platelet thrombus formation in eHUS is prevented by anti-MBL2


Autoři: R. I. Kushak aff001;  D. C. Boyle aff001;  I. A. Rosales aff001;  J. R. Ingelfinger aff001;  G. L. Stahl aff002;  M. Ozaki aff002;  R. B. Colvin aff001;  E. F. Grabowski aff001
Působiště autorů: Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, United States of America aff001;  Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, United States of America aff002;  Department of Emergency and Critical Care Medicine, St. Marianna University School of Medicine, Kawasaki, Japan aff003
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
doi: 10.1371/journal.pone.0220483

Souhrn

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+/+ 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.

Klíčová slova:

Electron microscopy – Fibrin – Glomeruli – Kidneys – Lectins – Mouse models – Platelets – Complement activation


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