Soluble endothelial cell molecules and circulating endothelial cells in patients with venous thromboembolism

Abstract

: To evaluate the plasma levels of soluble endothelial cell molecules in patients with venous thromboembolism (VTE) out of the acute phase as compared with healthy individuals. We also investigated the possible associations of the soluble endothelial cell molecules among them, as well as with other clinical and laboratory data, including the numbers of circulating endothelial cells (CEC), circulating endothelial progenitor cells (CEP), and CEC expressing activation-related [cluster of differentiation (CD)54 and CD62E] and procoagulant (CD142) markers. In total, 15 patients with VTE and 20 normal individuals were studied. The CEC and CEP were quantified and characterized by flow cytometry. The soluble molecules studied included P-selectin, E-selectin, intercellular cell adhesion molecule 1, vascular cell adhesion molecule 1 and tissue factor (ELISA), and von Willebrand factor antigen (immunoturbidimetry). VTE patients had significantly higher levels of vascular cell adhesion molecule 1 and von Willebrand factor antigen and lower levels of soluble E-selectin than controls. They also showed significantly higher numbers of CEC, as of activated/procoagulant CEC and lower numbers of CEP, compared with controls. We did not find any correlation between the levels of soluble molecules and the numbers of endothelial cell in circulation, but there was with several clinical and laboratory data in VTE patients. Our results would suggest that in VTE patients, the endothelium remains activated and in some hypercoagulable state. The levels of soluble endothelial cell molecules did not seem to be directly related to the numbers of CEC and CEP neither reflected the number of activated CEC, which may be because of the different function that surface and soluble molecules may have.

Biomarkers for the diagnosis of venous thromboembolism: D-dimer, thrombin generation, procoagulant phospholipid and soluble P-selectin

“…In contrast, a combination of sP-selectin cut-off ≤60 ng/mL combined with Wells score <2 resulted in a sensitivity of 99%, a specificity of 33% and a negative predictive value of 96% for the exclusion of DVT 10. More recently, Torres et al 24 reported a trend towards increased value of sP-selectin in 15 patients with previous VTE versus 20 normal individuals (median concentration 90 vs 72 ng/mL, respectively, p=0.099). However, in this study, blood samples were collected after the acute phase (at least a month after the last VTE).…”

The Blood Circulating Rare Cell Population. What Is It and What Is It Good for?

“…Furthermore, exhaustive enrichment may not always be necessary. Circulating EnC were simply identified, characterized, and quantified by flow cytometry using a stain-lyse-no-wash single-platform method [75]. Bethel et al [68] reported the isolation of endothelial cells without prior enrichment solely relying on RBC lysis and specialized fluorescence microscopy deemed the HD-CEC method.…”

“…The assay relied on positive selection of CD146+ cells followed by fluorescence microscopy. Hansmann et al [75]presented the "EPC capture chip" relying on positive selection of CD34+ cells and required only 200 µL blood for sample analysis. Due to variable marker expression in EPC and EnC, in particular with respect to CD34, positive selection introduces a bias inevitably leading to cell losses.…”

“…Apart from solid tissue cancer, circulating EnC and EPC have been associated with many other pathologies and this constitutes a serious pitfall when used as independent markers in cancer diagnosis. In the know of an incomplete list, endothelial elevation may occur upon ischemia, congestive heart failure, myocardial infarction (MI), sepsis, vascular trauma, hypertension, hyperlipidaemia, and hyperglycaemia, acute and chronic infection [94,95], sickle cell anemia, vasculitis, diabetes type II [96] deep vein thrombosis) [75,76], granulomatosis [5], pulmonary hypertension [74] or cirrhosis [97]. In inflammation, circulating EnC and EPC may interact with each other where inflammation activated circulating EnC induce EPC dysfuntion [5].…”

“…Awareness about enumeration inconsistencies and its remedies exist [244]. Torres et al [75] reported a median number of 504 circulating EnC. CEC were identified as CD45 negative, CD146 positive and CD133 negative cells.…”