Direkt zum Inhalt
Merck

Blocking neutrophil diapedesis prevents hemorrhage during thrombocytopenia.

The Journal of experimental medicine (2015-07-15)
Carina Hillgruber, Birgit Pöppelmann, Carsten Weishaupt, Annika Kathrin Steingräber, Florian Wessel, Wolfgang E Berdel, J Engelbert Gessner, Benoît Ho-Tin-Noé, Dietmar Vestweber, Tobias Goerge
ZUSAMMENFASSUNG

Spontaneous organ hemorrhage is the major complication in thrombocytopenia with a potential fatal outcome. However, the exact mechanisms regulating vascular integrity are still unknown. Here, we demonstrate that neutrophils recruited to inflammatory sites are the cellular culprits inducing thrombocytopenic tissue hemorrhage. Exposure of thrombocytopenic mice to UVB light provokes cutaneous petechial bleeding. This phenomenon is also observed in immune-thrombocytopenic patients when tested for UVB tolerance. Mechanistically, we show, analyzing several inflammatory models, that it is neutrophil diapedesis through the endothelial barrier that is responsible for the bleeding defect. First, bleeding is triggered by neutrophil-mediated mechanisms, which act downstream of capturing, adhesion, and crawling on the blood vessel wall and require Gαi signaling in neutrophils. Second, mutating Y731 in the cytoplasmic tail of VE-cadherin, known to selectively affect leukocyte diapedesis, but not the induction of vascular permeability, attenuates bleeding. Third, and in line with this, simply destabilizing endothelial junctions by histamine did not trigger bleeding. We conclude that specifically targeting neutrophil diapedesis through the endothelial barrier may represent a new therapeutic avenue to prevent fatal bleeding in immune-thrombocytopenic patients.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Hexadecyltrimethylammoniumbromid, ≥98%
Sigma-Aldrich
Formaldehyd -Lösung, for molecular biology, 36.5-38% in H2O
Sigma-Aldrich
Hexadecyltrimethylammoniumbromid, for molecular biology, ≥99%
Sigma-Aldrich
Formaldehyd -Lösung, ACS reagent, 37 wt. % in H2O, contains 10-15% Methanol as stabilizer (to prevent polymerization)
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidin, ≥99%
Sigma-Aldrich
Hexadecyltrimethylammoniumbromid, BioXtra, ≥99%
Sigma-Aldrich
Formaldehyd -Lösung, for molecular biology, BioReagent, ≥36.0% in H2O (T)
Sigma-Aldrich
N,O-Bis(trimethylsilyl)acetamid, synthesis grade, ≥95%
Sigma-Aldrich
Calcein, Used for the fluorometric determination of calcium and EDTA titration of calcium in the presence of magnesium.
Sigma-Aldrich
Phenylessigsäure, 99%
Sigma-Aldrich
Histamin, ≥97.0%
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidin, ≥98% (TLC)
Sigma-Aldrich
Hexadecyltrimethylammoniumbromid, BioUltra, for molecular biology, ≥99.0% (AT)
Sigma-Aldrich
Formaldehyd -Lösung, meets analytical specification of USP, ≥34.5 wt. %
Sigma-Aldrich
Phenylessigsäure, ≥99%, FCC, FG
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidin, ≥98.0% (NT)
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidin, tablet, 1 mg substrate per tablet
Sigma-Aldrich
Fluo-3, suitable for fluorescence, ~70%
SAFC
Hexadecyltrimethylammoniumbromid
Sigma-Aldrich
Formaldehyd-12C -Lösung, 20% in H2O, 99.9 atom % 12C