Prothrombotic effects of tumor necrosis factor alpha in vivo are amplified by the absence of TNF-alpha receptor subtype 1 and require TNF-alpha receptor subtype 2Report as inadecuate

Prothrombotic effects of tumor necrosis factor alpha in vivo are amplified by the absence of TNF-alpha receptor subtype 1 and require TNF-alpha receptor subtype 2 - Download this document for free, or read online. Document in PDF available to download.

Arthritis Research and Therapy

, 14:R225

First Online: 18 October 2012Received: 16 February 2012Revised: 31 August 2012Accepted: 05 October 2012


IntroductionElevated serum levels of the proinflammatory cytokine tumor necrosis factor alpha TNFα correlate with an increased risk for atherothrombotic events and TNFα is known to induce prothrombotic molecules in endothelial cells. Based on the preexisting evidence for the impact of TNFα in the pathogenesis of autoimmune disorders and their known association with an acquired hypercoagulability, we investigated the effects of TNFα and the role of the TNF receptor subtypes TNFR1 and TNFR2 for arteriolar thrombosis in vivo.

MethodsArteriolar thrombosis and platelet-rolling in vivo were investigated in wildtype, TNFR1-, TNFR2- and TNFR1-R2- C57BL-6 mice using intravital microscopy in the dorsal skinfold chamber microcirculation model. In vitro, expression of prothrombotic molecules was assessed in human endothelial cells by real-time PCR and flow cytometry.

ResultsIn wildtype mice, stimulation with TNFα significantly accelerated thrombotic vessel occlusion in vivo upon ferric chloride injury. Arteriolar thrombosis was much more pronounced in TNFR1- animals, where TNFα additionally led to increased platelet-endothelium-interaction. TNFα dependent prothrombotic effects were not observed in TNFR2- and TNFR1-R2- mice. In vitro, stimulation of human platelet rich plasma with TNFα did not influence aggregation properties. In human endothelial cells, TNFα induced superoxide production, p-selectin, tissue factor and PAI-1, and suppressed thrombomodulin, resulting in an accelerated endothelial dependent blood clotting in vitro. Additionally, TNFα caused the release of soluble mediators by endothelial cells which induced prothrombotic and suppressed anticoagulant genes comparable to direct TNFα effects.

ConclusionsTNFα accelerates thrombus formation in an in vivo model of arteriolar thrombosis. Its prothrombotic effects in vivo require TNFR2 and are partly compensated by TNFR1. In vitro studies indicate endothelial mechanisms to be responsible for prothrombotic TNFα effects. Our results support a more selective therapeutic approach in anticytokine therapy favouring TNFR2 specific antagonists.

AbbreviationsFITCfluorescein isothiocyanate

GAPDHglyceraldehyde-3-phosphate dehydrogenase

HMEChuman microvascular endothelial cell

HUVEChuman umbilical vein endothelial cell

NF-κBnuclear factor-kappa-B

PAI-1plasminogen activator inhibitor 1

PCRpolymerase chain reaction

PRPplatelet-rich plasma

PVWIplatelet-vessel wall interaction

RArheumatoid arthritis


siRNAsmall interfering RNA

tmTNFαtransmembrane tumor necrosis factor alpha

TNFαtumor necrosis factor alpha

TNFR1tumor necrosis factor alpha receptor subtype 1

TNFR2tumor necrosis factor alpha receptor subtype 2

TRAPthrombin receptor-activating protein


Electronic supplementary materialThe online version of this article doi:10.1186-ar4064 contains supplementary material, which is available to authorized users.

Joachim Pircher, Monika Merkle, Markus Wörnle contributed equally to this work.

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Author: Joachim Pircher - Monika Merkle - Markus Wörnle - Andrea Ribeiro - Thomas Czermak - Yvonn Stampnik - Hanna Mannell - Marku


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