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• W1536869080 abstract "Previously regarded as inert cell debris, in recent years microparticles have been incriminated in the pathophysiology of different diseases. Especially, microparticle‐associated tissue factor (MPTF)‐activity is associated with the clinical manifestation of thrombosis in cancer patients [1Khorana A.A. Francis C.W. Menzies K.E. Wang J.G. Hyrien O. Hathcock J. Mackman N. Taubman M.B. Plasma tissue factor may be predictive of venous thromboembolism in pancreatic cancer.J Thromb Haemost. 2008; 6: 1983-5Crossref PubMed Scopus (242) Google Scholar, 2Manly D.A. Wang J. Glover S.L. Kasthuri R. Liebman H.A. Key N.S. Mackman N. Increased microparticle tissue factor activity in cancer patients with venous thromboembolism.Thromb Res. 2010; 125: 511-2Abstract Full Text Full Text PDF PubMed Scopus (175) Google Scholar, 3Tesselaar M.E. Romijn F.P. van der Linden I.K. Bertina R.M. Osanto S. Microparticle‐associated tissue factor activity in cancer patients with and without thrombosis.J Thromb Haemost. 2009; 7: 1421-3Crossref PubMed Scopus (0) Google Scholar, 4Tesselaar M.E. Romijn F.P. van der Linden I.K. Prins F.A. Bertina R.M. Osanto S. Microparticle‐associated tissue factor activity: a link between cancer and thrombosis?.J Thromb Haemost. 2007; 5: 520-7Crossref PubMed Scopus (0) Google Scholar]. High MPTF‐activity has also been reported in patients with disseminated intravascular coagulation (DIC) in gram‐negative sepsis [5Nieuwland R. Berckmans R.J. McGregor S. Boing A.N. Romijn F.P. Westendorp R.G. Hack C.E. Sturk A. Cellular origin and procoagulant properties of microparticles in meningococcal sepsis.Blood. 2000; 95: 930-5Crossref PubMed Google Scholar]. Aras et al. [6Aras O. Shet A. Bach R.R. Hysjulien J.L. Slungaard A. Hebbel R.P. Escolar G. Jilma B. Key N.S. Induction of microparticle‐ and cell‐associated intravascular tissue factor in human endotoxemia.Blood. 2004; 103: 4545-53Crossref PubMed Scopus (0) Google Scholar] have injected healthy male volunteers with Escherichia coli lipopolysaccharide (LPS) to assess the time course of both MPTF and whole blood TF. They found a transient increase in non‐platelet MPTF procoagulant activity (PCA) and a more prolonged increase of whole blood TF PCA, which they attributed to monocyte‐bound TF. In their experiments, microparticles were isolated using a monoclonal antibody recognizing an unidentified antigen on fibroblasts, monocytes, smooth muscle cells, granulocytes and endothelial cells. We repeated their experiments with a different assay in an extensively documented in vivo human endotoxemia model in order to relate the dynamics of procoagulant microparticles to earlier published parameters of inflammation, coagulation and endothelial activation [7de Kruif M.D. Lemaire L.C. Giebelen I.A. van Zoelen M.A. Pater J.M. van den Pangaart P.S. Groot A.P. de Vos A.F. Elliott P.J. Meijers J.C. Levi M. van der Poll T. Prednisolone dose‐dependently influences inflammation and coagulation during human endotoxemia.J Immunol. 2007; 178: 1845-51Crossref PubMed Google Scholar, 8Lemaire L.C. de Kruif M.D. Giebelen I.A. van Zoelen M.A. van’t Veer C. van der Poll T. Differential dose‐dependent effects of prednisolone on shedding of endothelial adhesion molecules during human endotoxemia.Immunol Lett. 2008; 121: 93-6Crossref PubMed Scopus (13) Google Scholar]. Briefly, eight healthy male volunteers received a single bolus of E. coli LPS (4 ng kg−1 body weight; lot G, US Pharmacopeia, Rockville, MD, USA) intravenously at t = 0 h. Citrated plasma was obtained at several time‐points during 24 h after LPS administration by immediate centrifugation and stored at −80 °C until assayed. All subjects provided written consent. Microparticles were isolated from platelet‐poor plasma through high speed centrifugation at 18 890 × g with minimum brake. MPTF‐activity was subsequently measured as FVII and TF‐dependent FXa formation using a previously validated method and expressed as fM Xa min−1 [4Tesselaar M.E. Romijn F.P. van der Linden I.K. Prins F.A. Bertina R.M. Osanto S. Microparticle‐associated tissue factor activity: a link between cancer and thrombosis?.J Thromb Haemost. 2007; 5: 520-7Crossref PubMed Scopus (0) Google Scholar, 9Garcia Rodriguez P. Eikenboom H.C. Tesselaar M.E. Huisman M.V. Nijkeuter M. Osanto S. Bertina R.M. Plasma levels of microparticle‐associated tissue factor activity in patients with clinically suspected pulmonary embolism.Thromb Res. 2010; 126: 345-9Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar]. Ultimately, only seven of eight volunteers (mean age ± SD 25.7 ± 5.2) were analyzed for MPTF‐activity due to insufficient plasma for microparticle isolation. During endotoxemia, mean MPTF‐activity ± SD transiently rose from 47 ± 41 fM Xa min−1 at baseline (range 0–123) to a peak value of 309 ± 267 fM Xa min−1 at t = 4 h (range 52–710) in accordance with the kinetics previously reported by Aras et al. (Fig. 1A). We similarly observed a large interindividual variation in MPTF‐response. Two subjects did not respond with an increase in MPTF‐activity after LPS stimulation, possibly reflecting the ‘high’ and ‘low’ LPS responder phenomenon [10Østerud B. The high responder phenomenon: enhancement of LPS induced tissue factor activity in monocytes by platelets and granulocytes.Platelets. 1995; 6: 119-25Crossref PubMed Scopus (0) Google Scholar]. This phenomenon describes the interindividual platelet and plasma‐dependent diversity in the levels of functional TF on monocytes in response to LPS. Consequently, levels of functional TF on generated monocyte‐derived microparticles may also vary, resulting in different levels of MPTF‐activity. Interestingly, one subject responded with an early peak in MPTF‐activity at t = 1 h, while having normal cytokine levels, C‐reactive protein and procalcitonine at baseline. Another subject, who had the most serious clinical response to endotoxin, had the highest levels of pro‐inflammatory cytokines (TNFα, IL‐6, IL‐8 were 5024, >10 000, 3586 pg mL−1 respectively) as well as MPTF‐activity (710 fM Xa min−1). Monocyte activation and recruitment as reflected in decreasing monocyte counts at t = 0.5 h preceded the formation of procoagulant microparticles (Fig. 1B). Comparison of our data with those previously obtained in essentially the same human endotoxemia model [11Franco R.F. de Jonge E. Dekkers P.E. Timmerman J.J. Spek C.A. van Deventer S.J. van Deursen P. van Kerkhoff L. van Gemen B. ten Cate H. van der Poll T. Reitsma P.H. The in vivo kinetics of tissue factor messenger RNA expression during human endotoxemia: relationship with activation of coagulation.Blood. 2000; 96: 554-9Crossref PubMed Google Scholar] shows that the peak in MPTF‐activity follows approximately 1 h after the peak in the TF mRNA/monocyte ratio previously reported, supporting the hypothesis that procoagulant microparticles are shedded from activated monocytes. MPTF‐activity generally followed TNFα secretion (peak at t = 1.5 h) [7de Kruif M.D. Lemaire L.C. Giebelen I.A. van Zoelen M.A. Pater J.M. van den Pangaart P.S. Groot A.P. de Vos A.F. Elliott P.J. Meijers J.C. Levi M. van der Poll T. Prednisolone dose‐dependently influences inflammation and coagulation during human endotoxemia.J Immunol. 2007; 178: 1845-51Crossref PubMed Google Scholar], but the interval between the two peaks varied, suggesting that MPTF generation in this model is mainly induced by LPS and not by TNFα. The increase in MPTF‐activity did, however, coincide with that of the pro‐inflammatory cytokines IL‐6 and IL‐8 and the anti‐inflammatory cytokine IL‐10 both in early and late MPTF‐responders (Fig. 1C,D; only IL‐6 shown). The increase in MPTF‐activity furthermore coincided with the increase in thrombin‐antithrombin complex (TATc) and prothrombin fragments 1 and 2 (F1+2), indicating activation of the coagulation system (Fig. 1E). The increase in soluble TF antigen (sTF; ELISA, American Diagnostics, Greenwich, CT, USA) preceded MPTF‐activity by approximately 2 h and sTF levels returned to baseline at the peak in MPTF‐activity (Fig. 1A). Total plasma sTF may consist of MPTF, truncated TF and alternatively spliced TF. As yet, it is not known which form of intravascular TF increases after LPS stimulation. Also it cannot be excluded that the ELISA is sensitive to some cross‐reactive substance. Considering that the rise and fall of sTF forms a mirror image of the decrease and increase in monocyte count (compare Fig. 1A,B), we hypothesize that sTF is of monocytic origin, mostly due to proteolytic cleavage of TF from the cell membrane and to a minor extent due to shedding of microparticles expressing TF. Although less probable, decryption of non‐functional tissue factor on microparticles may also contribute to the rise in MPTF‐activity. Even though LPS can also induce shedding of TF bearing microparticles from endothelial cells in vitro [12Del Turco S. Basta G. Lazzerini G. Evangelista M. Rainaldi G. Tanganelli P. Camera M. Tremoli E. De Caterina R. Parallel decrease of tissue factor surface exposure and increase of tissue factor microparticle release by the n‐3 fatty acid docosahexaenoate in endothelial cells.Thromb Haemost. 2007; 98: 210-9Crossref PubMed Scopus (0) Google Scholar], their existence in vivo is a matter of debate. In our study, levels of the endothelial activation markers von Willebrand factor antigen (VWF Ag) and soluble E‐selectin (sE‐selectin) increased slightly later than the MPTF‐activity and remained relatively high (Fig. 1F), rendering an endothelial origin of MPTF‐activity unlikely. In conclusion, we confirmed the data of Aras et al. on the kinetics of procoagulant microparticles in the in vivo human endotoxemia model using a different method for the measurement of MPTF‐activity. Additionally, we demonstrated that the levels of MPTF‐activity increased concurrently with markers of inflammation and coagulation, but decreased before endothelial activation markers peaked. We showed variation in the levels of MPTF‐activity with two non‐responders, as well as in the onset of MPTF‐activity with one early and four late responders. Whether MPTF‐activity merely represents a reflection of the number of activated or apoptotic cells expressing functional TF or in itself contributes to DIC remains elusive. Further research in patients with gram‐negative sepsis is warranted in order to establish the disease severity in the non‐, early and late MPTF‐responders. The authors state that they have no conflict of interest. This work was supported by the Dutch Cancer Society (KWF UL 2006‐3618)." @default.
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• W1536869080 title "Microparticles expressing tissue factor are concurrently released with markers of inflammation and coagulation during human endotoxemia" @default.
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