Systemic coagulation activation. a Vascular endothelial growth factor …

Systemic coagulation activation. a Vascular endothelial growth factor (VEGF) antigen levels were measured in plasma or PBMC culture supernatants by ELISA. b Based on VEGF antigen levels in culture super natants, AML patients were grouped into those with PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/14960617 (>40 pg/mL) and those without (40 pg/mL) appreciable VEGF production. No significant differences were found with regard to TF PCA levels in intact or disrupted PBMCs. c Levels of cellfree plasma DNA were quantified in patients and controls as described under the section "Methods". Plasma for DNA quantification was not available from four patients with AML. d Based on levels of cellfree plasma DNA, patients were grouped into those with DNA levels greater or equal or less than N,N'-Dimethyl-N-[2-(methylamino)ethyl]ethylenediamine the cutoff value of 110 ng/mL defined by the 95th percentile within the control group. e In a modified analysis, the seven AML patients with DIC and TF PCA levels of intact PBMCs exceeding 2,500 AU/106 cells were excluded. P values are according to Mann hitney U test.Importantly, DIC meo.v19.25901 was not restricted to APL patients, but also occurred in seven other patients, three of whom had acute (myelo)monocytic leukemia. To our knowledge, this is one of the largest prospective studies so far specifically addressing the determinants of systemic coagulation activation in newly diagnosed AML. The increase in PBMC-associated TF PCA Methyl 4-chloro-5-fluoroanthranilate following cell disruption is consistent with the concept of cryptic TF that can be de-encrypted upon cellular activation or induction of apoptosis/necrosis [24]. Previous studies have clearly shown that cryptic TF is entirely cell surface-expressed on (myelo)monocytic cells and not liberated from intracellular storage pools upon cell lysis[25?7]. Instead, negatively charged phospholipids (i.e. phosphatidylserine) exposed on the outer membrane leaflet act synergistically PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/9638577 with thiol-disulfide exchange reactions to activate surface-expressed TF [28?0]. In this regard, our findings provide circumstantial clinical evidence that membrane alterations associated with apoptosis/necrosis indeed contribute to cellular TF activation in vivo, because serum LDH levels, a surrogate marker for leukemic cell turnover, significantly correlated with plasma D-dimer. Systemic coagulation activation in AML is thus, at least to a substantial extent, determined by the following three factors: specific upregulation of cellular TF PCA following leukemic transformation ofDicke et al. Exp Hematol Oncol (2015) 4:Page 10 ofaCumulative survival ( )bCumulative survival ( )VEGF 40 pg/mL (n=28) VEGF >40 pg/mL (n=17)P=0.0 0 12 24 36 48 60Follow-up (months)Follow-up (months)cCumulative survival ( )TF PCA >AU/cells (n=19)Cumulative survival ( )TF PCA 735 AU/106 cells (n=50)dTF PCA 293 AU/106 cells (n=35) TF PCA >293 AU/106 cells (n=34)P=0.P=0.0 0 12 24 36 48 600 0 12 24 36 48 60Follow-up (months)Follow-up (months)eCumulative survival ( )DIC (n=11)Cumulative survival ( )no DIC (n=58)fno DIC (n=36) DIC (n=8)P=0.P=0.0 0 6 12 18 24 300 0 12 24 36 48 60Follow-up (months)Follow-up (months)Fig. 6 Impact of VEGF production, TF PCA expression and DIC evolution on overall survival. a Overall survival of the total AML patient cohort (n = 69). The dashed lines indicate the 95 confidence interval. b Overall survival of AML patients grouped according to VEGF antigen levels in cul ture supernatants. c, d Overall survival of AML patients grouped according to TF PCA levels of intact PBMCs. Cutoff values for TF PCA were defined as the 95th percentile wit.