Measurement of tissue factor messenger RNA levels in leukocytes from patients in hypercoagulable state caused by several underlying diseases

2003 ◽  
Vol 89 (04) ◽  
pp. 660-665 ◽  
Author(s):  
Tomohiro Sase ◽  
Hideo Wada ◽  
Junji Nishioka ◽  
Yasunori Abe ◽  
Esteban Gabazza ◽  
...  
Keyword(s):  
Healthy Volunteers ◽  
Tissue Factor ◽  
Messenger Rna ◽  
Solid Cancer ◽  
Mrna Levels ◽  
Factor X ◽  
Reactive Protein ◽  
Preliminary Study ◽  
Underlying Diseases

SummaryIn a preliminary study, we have demonstrated that tissue factor (TF) is immunohistochemically stained in monocytes from patients with disseminated intravascular coagulation (DIC) but not from healthy volunteers, and that leukocytes from DIC patients induce enhanced activated factor X (FXa) generation in the presence of a mixture of FVIIa, FX and Ca2+. Then, TF mRNA levels in leukocytes were measured to evaluate the role of TF in the pathophysiology of various diseases. TF mRNA levels in leukocytes were low in healthy volunteers but they were significantly increased in various diseases, especially in patients with infectious diseases, solid cancer, anemd hematopoietic tumors. TF mRNA levels in leukocytes were significantly higher in patients with high levels of C reactive protein (CRP) than in those with low CRP. TF mRNA levels were significantly higher in patients with DIC than in those without DIC. TF mRNA levels were well correlated with TF antigens in plasma and leukocytes. These findings suggest that the expression of TF mRNA in leukocytes is increased in various diseases and that this may play an important role in hypercoagulability or DIC.

scholarly journals In Hemophilia Α Plasma Treated with Emicizumab, Factor IX Activation By Factor VIIa Drives Thrombin Generation

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 17-17
Author(s):  
Dougald Monroe ◽  
Mirella Ezban ◽  
Maureane Hoffman
Keyword(s):  
Factor Viii ◽  
Tissue Factor ◽  
Plasma Levels ◽  
Thrombin Generation ◽  
Hemophilia A ◽  
Factor Viia ◽  
Factor Ix ◽  
Factor X ◽  
Dose Dependent

Background.Recently a novel bifunctional antibody (emicizumab) that binds both factor IXa (FIXa) and factor X (FX) has been used to treat hemophilia A. Emicizumab has proven remarkably effective as a prophylactic treatment for hemophilia A; however there are patients that still experience bleeding. An approach to safely and effectively treating this bleeding in hemophilia A patients with inhibitors is recombinant factor VIIa (rFVIIa). When given at therapeutic levels, rFVIIa can enhance tissue factor (TF) dependent activation of FX as well as activating FX independently of TF. At therapeutic levels rFVIIa can also activate FIX. The goal of this study was to assess the role of the FIXa activated by rFVIIa when emicizumab is added to hemophilia A plasma. Methods. Thrombin generation assays were done in plasma using 100 µM lipid and 420 µM Z-Gly-Gly-Arg-AMC with or without emicizumab at 55 µg/mL which is the clinical steady state level. The reactions were initiated with low (1 pM) tissue factor (TF). rFVIIa was added at concentrations of 25-100 nM with 25 nM corresponding to the plasma levels achieved by a single clinical dose of 90 µg/mL. To study to the role of factor IX in the absence of factor VIII, it was necessary to create a double deficient plasma (factors VIII and IX deficient). This was done by taking antigen negative hemophilia B plasma and adding a neutralizing antibody to factor VIII (Haematologic Technologies, Essex Junction, VT, USA). Now varying concentrations of factor IX could be reconstituted into the plasma to give hemophilia A plasma. Results. As expected, in the double deficient plasma with low TF there was essentially no thrombin generation. Also as expected from previous studies, addition of rFVIIa to double deficient plasma gave a dose dependent increase in thrombin generation through activation of FX. Interestingly addition of plasma levels of FIX to the rFVIIa did not increase thrombin generation. Starting from double deficient plasma, as expected emicizumab did not increase thrombin generation since no factor IX was present. Also, in double deficient plasma with rFVIIa, emicizumab did not increase thrombin generation. But in double deficient plasma with FIX and rFVIIa, emicizumab significantly increased thrombin generation. The levels of thrombin generation increased in a dose dependent fashion with higher concentrations of rFVIIa giving higher levels of thrombin generation. Conclusion. Since addition of FIX to the double deficient plasma with rFVIIa did not increase thrombin generation, it suggests that rFVIIa activation of FX is the only source of the FXa needed for thrombin generation. So in the absence of factor VIII (or emicizumab) FIX activation does not contribute to thrombin generation. However, in the presence of emicizumab, while rFVIIa can still activate FX, FIXa formed by rFVIIa can complex with emicizumab to provide an additional source of FX activation. Thus rFVIIa activation of FIX explains the synergistic effect in thrombin generation observed when combining rFVIIa with emicizumab. The generation of FIXa at a site of injury is consistent with the safety profile observed in clinical use. Disclosures Monroe: Novo Nordisk:Research Funding.Ezban:Novo Nordisk:Current Employment.Hoffman:Novo Nordisk:Research Funding.

scholarly journals Endotoxin-induced tissue factor messenger RNA in human monocytes is negatively regulated by a cyclic AMP-dependent mechanism

Blood ◽  
1993 ◽  
Vol 81 (4) ◽  
pp. 973-979 ◽  
Author(s):  
V Ollivier ◽  
S Houssaye ◽  
C Ternisien ◽  
A Leon ◽  
H de Verneuil ◽  
...  
Keyword(s):  
Cyclic Amp ◽  
Tissue Factor ◽  
Messenger Rna ◽  
Factor Viia ◽  
Functional Expression ◽  
Mrna Levels ◽  
Proteolytic Activation ◽  
Protein Levels ◽  
Tf Gene ◽  
Camp Levels

Abstract Tissue factor (TF) is a transmembrane receptor that serves as the major cofactor for factor VIIa-catalyzed proteolytic activation of factors IX and X. In response to bacterial lipopolysaccharide (LPS), monocytes transcribe, synthesize, and express TF on their surface, thereby conveying to activated monocytes the ability to initiate the blood coagulation protease cascades. Agents that elevate cellular cyclic AMP (cAMP) inhibit the functional expression of TF by LPS-stimulated monocytes. In this study, we investigated the mechanism of this suppression. Northern blot analysis of total RNA from LPS-stimulated monocytes showed a concentration-dependent decrease in TF messenger RNA (mRNA) levels in response to dibutyryl-cAMP (dBt-cAMP). TF mRNA and procoagulant activity were inhibited as early as 1 hour after the addition of dBt-cAMP and the inhibition persisted through 4 hours. Suppression of specific mRNA abundance was also observed with agents, including forskolin and iso-butyl-methyl-xanthine (IBMX), that increase cAMP levels by independent mechanisms. Flow immunocytometric analysis confirmed that cell-surface TF protein levels declined in parallel with TF functional activity. The rate of decay of TF mRNA after the arrest of transcription by actinomycin D was not altered by the addition of dBt-cAMP, IBMX, or forskolin, thus excluding effects on TF mRNA stability. We conclude that elevated cAMP levels suppress TF mRNA by reducing the rate of TF gene transcription.

Tissue factor messenger RNA levels in leukocytes compared with tissue factor antigens in plasma from patients in hypercoagulable state caused by various diseases

2004 ◽  
Vol 92 (07) ◽  
pp. 132-139 ◽  
Author(s):  
Tomohiro Sase ◽  
Yuko Kamikura ◽  
Toshihiro Kaneko ◽  
Yasunori Abe ◽  
Junji Nishioka ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Plasma Levels ◽  
Messenger Rna ◽  
Healthy Subjects ◽  
Flow Cytometric Analysis ◽  
Antigen Expression ◽  
Normal Subjects ◽  
Hypercoagulable State ◽  
Mrna Levels ◽  
All Diseases

SummaryWe compared the levels of tissue factor (TF) mRNA in leukocytes with plasma TF antigens of patients in hypercoagulable state caused by various diseases. Flow cytometric analysis showed absence of TF antigen expression on neutrophils and monocytes in healthy subjects but strong expression in both cell types of patients with infections. TF mRNA levels in leukocytes were low in healthy subjects but they were significantly elevated in patients with underlying diseases of disseminated intravascular coagulation (DIC), especially in acute myeloid leukaemia (AML) and infections. TF mRNA levels in leukocytes were significantly high in patients with all diseases except those with thrombosis, and plasma TF antigen levels were significantly high in all diseases. TF mRNA in leukocytes and plasma TF antigen levels were significantly high in patients with overt-DIC, and TF mRNA/antigen ratio was significantly high in patients with overt-DIC. In patients with solid cancers, TF mRNA and TF mRNA/antigen ratio were significantly higher in patients with metastases than those without. TF mRNA levels in leukocytes and plasma levels of TF antigen did not correlate in normal subjects and all patients, but they tended to be correlated in patients with AML, infections or overt-DIC. Our analysis suggests that TF expression in leukocytes plays an important role in various diseases but the expression level does not always correlate with plasma levels of TF antigen.

scholarly journals The effect of interleukin-1 on C-reactive protein expression in Hep3B cells is exerted at the transcriptional level

1995 ◽  
Vol 310 (1) ◽  
pp. 143-148 ◽  
Author(s):  
D Zhang ◽  
S L Jiang ◽  
D Rzewnicki ◽  
D Samols ◽  
I Kushner
Keyword(s):  
Interleukin 6 ◽  
Interleukin 1 ◽  
Kinetic Studies ◽  
Acute Phase Reactant ◽  
Transcriptional Level ◽  
Mrna Levels ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Hep3b Cells

The combination of interleukin 6 (IL-6) and interleukin 1 (IL-1) synergistically induces the human acute-phase reactant, C-reactive protein (CRP) in Hep3B cells. While previous studies have indicated that IL-6 induces transcription of CRP, the mode of action of IL-1 has not been clearly defined. It has been suggested that the effect of IL-1 might be post-transcriptional, exerted through the 5′-untranslated region (5′-UTR). To evaluate the role of IL-1 in CRP gene expression, we studied the effects of interleukin-6 (IL-6) and interleukin-1 beta (IL-1 beta) on both the endogenous CRP gene and on transfected CRP-CAT constructs in Hep3B cells. In kinetic studies of the endogenous CRP gene, IL-1 beta alone had no effect on CRP mRNA levels, but when added to IL-6, synergistically enhanced both CRP mRNA levels and transcription, as determined by Northern-blot analyses and nuclear run-on studies. IL-6 alone and the combination of [IL-1 beta + IL-6] each induced increases in mRNA levels roughly comparable with observed increases in transcription. These findings indicate that the effect of IL-1 beta on CRP expression is exerted largely at the transcriptional level in this system. This conclusion was confirmed by studies in Hep3B cells transiently transfected with CRP-CAT constructs, each containing 157 bp of the CRP 5′-flanking region but differing in the length of the 5′-UTR from 104 bp to 3 bp. All constructs responded in the same way; IL-6, but not IL-1 beta, induced significant chloramphenicol acetyltransferase (CAT) expression which was synergistically enhanced 2- to 3-fold by IL-1 beta. These results indicate that IL-1 beta stimulates transcriptional events in the presence of IL-6 and that the upstream 157 bases of the CRP promoter contain elements capable of both IL-6 induction and the synergistic effect of IL-1 beta on transcription.

scholarly journals Retinoic acid counteracts both the downregulation of thrombomodulin and the induction of tissue factor in cultured human endothelial cells exposed to tumor necrosis factor

Blood ◽  
1992 ◽  
Vol 80 (10) ◽  
pp. 2556-2562 ◽  
Author(s):  
H Ishii ◽  
S Horie ◽  
K Kizaki ◽  
M Kazama
Keyword(s):  
Endothelial Cells ◽  
Retinoic Acid ◽  
Tumor Necrosis Factor ◽  
Tissue Factor ◽  
Inflammatory Cytokines ◽  
Messenger Rna ◽  
Tumor Necrosis ◽  
Tnf Alpha ◽  
Mrna Levels ◽  
Necrosis Factor

Abstract Inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) shift the hemostatic balance of endothelial cell surfaces in favor of prothrombotic properties by downregulating thrombomodulin (TM) and inducing tissue factor (TF) expression. We investigated the effects of retinoic acid (RA) on the prothrombotic properties of cultured umbilical vein endothelial cells exposed to TNF-alpha. The approximate 50% downregulation of TM antigen and cofactor activity induced by TNF- alpha (10 U/mL for 24 hours) was completely prevented when the cells were coincubated with both TNF-alpha and 10 mumol/L RA. In accordance with changes in cell surface TM antigen levels, the 70% decrease in TM messenger RNA (mRNA) induced by TNF-alpha was also prevented by 10 mumol/L RA. TNF-alpha induced TF activity of lysed cells (100-fold greater than untreated controls), an effect prevented when the cells were coincubated with both the TNF-alpha and 10 mumol/L RA. The 34-fold increase in TF mRNA levels induced by TNF-alpha (10 U/mL for 3 hours) was only two-fold in the presence of both TNF-alpha and RA. The effects of RA on the regulation of TM and TF expression in the cells exposed to TNF-alpha was dose-dependent from 0.01 to 10 mumol/L RA. The present results suggest that RA may affect on the mRNA level to alter TM and TF expression, effectively counteracting expression of prothrombotic properties of endothelial cells induced by inflammatory cytokines such as TNF-alpha.

scholarly journals Methylation and Gene Expression of BCAT1 and IKZF1 in Colorectal Cancer Tissues

2018 ◽  
Vol 12 ◽  
pp. 117955491877506 ◽  
Author(s):  
Maher Jedi ◽  
Graeme P Young ◽  
Susanne K Pedersen ◽  
Erin L Symonds
Keyword(s):  
Gene Expression ◽  
Colorectal Cancer ◽  
Messenger Rna ◽  
Stage Iv ◽  
Mrna Levels ◽  
Polymerase Chain ◽  
Cancer Tissues ◽  
The Relationship ◽  
Neoplastic Tissues

The genes BCAT1 and IKZF1 are hypermethylated in colorectal cancer (CRC), but little is known about how this relates to gene expression. This study assessed the relationship between methylation and gene expression of BCAT1 and IKZF1 in CRC and adjacent non-neoplastic tissues. The tissues were obtained at surgery from 36 patients diagnosed with different stages of CRC (stage I n = 8, stage II n = 13, stage III n = 10, stage IV n = 5). Methylated BCAT1 and IKZF1 were detected in 92% and 72% CRC tissues, respectively, with levels independent of stage ( P > .05). In contrast, only 31% and 3% of non-neoplastic tissues were methylated for BCAT1 and IKZF1, respectively ( P < .001). The IKZF1 messenger RNA (mRNA) expression was significantly lower in the cancer tissues compared with that of non-neoplastic tissues, whereas the BCAT1 mRNA levels were similar. The latter may be due to the BCAT1 polymerase chain reaction assay detecting more than 1 mRNA transcript. Further studies are warranted to establish the role of the epigenetic silencing of IKZF1 in colorectal oncogenesis.

State-of-the-Art Review : Role of Antithrombin and Tissue Factor Pathway Inhibitor in the Control of Thrombosis and Mediation of Heparin Action

1996 ◽  
Vol 2 (1) ◽  
pp. 1-6 ◽  
Author(s):  
Nina Iverson ◽  
Ulrich Abildgaard
Keyword(s):  
Tissue Factor ◽  
Tissue Factor Pathway Inhibitor ◽  
State Of The Art ◽  
Factor Viia ◽  
Heparin Therapy ◽  
Factor X ◽  
Factor Ixa ◽  
Tissue Factor Pathway ◽  
Blocking Antibodies

Deficiency of any of the two coagulation in hibitors antithrombin (AT) and tissue factor pathway in hibitor (TFPI) lowers the resistance to thrombosis. He reditary deficiency of AT leads to a high risk of throm bosis, which occasionally responds poorly to heparin therapy. Experimental deficiency of TFPI lowers the re sistance to infusion of both tissue factor and endotoxin, both regarding microvascular thrombosis and fatality. Administration of either AT or TFPI protects against mi cro- and macrovascular thrombosis. Injection of heparin and some other glycosaminoglycans releases intima bound TFPI to the blood. Heparin accelerates the inhib itory effects of both inhibitors, in particular the effect of AT. The influence of the two inhibitors on the various anticoagulant reactions have been studied using blocking antibodies. It is suggested that the anticoagulant and an tithrombotic effects of heparin are mainly mediated by the accelerated inactivation of thrombin, factor IXa and factor X by AT, and augmented inactivation of tissue factor-factor VIIa by TFPI released to the blood.

scholarly journals Role of N6-Methyladenosine (m6A) RNA Modification in Multiple Myeloma

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 5601-5601
Author(s):  
Christian Bach ◽  
Magdalena Leffler ◽  
Cindy Flamann ◽  
Jan Kronke ◽  
Dimitrios Mougiakakos ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Lines ◽  
Messenger Rna ◽  
Plasma Cells ◽  
Conflicts Of Interest ◽  
Stop Codon ◽  
Mrna Levels ◽  
Molecular Abnormalities ◽  
Gene Expression Control

Abstract Multiple myeloma (MM) is considered a chronic and incurable disease due to its highly complex and heterogeneous molecular abnormalities. In recent years, integrating proteasome inhibitors and immunomodulatory drugs into MM frontline therapy has significantly improved treatment efficacy with a median overall survival (OS) being prolonged from 3-4 to 7 years. Despite this progress, patients refractory to the aforementioned agent classes display a median OS of only 9 months. Thus, the clinical necessity for developing novel therapeutic alternative approaches is self-evident. Methylation of N6-adenosine (m6A) is known to be important for diverse biological processes including gene expression control, translation of protein, and messenger RNA (mRNA) splicing. m6A regulatory enzymes consist of "writers" METTL3 and METTL14, "readers" YTHDF1 and YTHDF2, and "erasers" FTO and ALKBH5. However, the functions of m6A mRNA modification and the specific role of these enzymes in MM remain unknown. Here we report that METTL3, a key component of the m6A methyltransferase complex, is highly expressed in MM cell lines and in isolated patient's MM cells. In contrast, we found no significant differences in the expression of the m6A demethylases FTO and ALKBH5. Accordingly, compared to plasma cells from healthy donors, global PolyA+ RNA showed a significant increase in m6A content in patient's MM plasma cells. In MM cell lines, global m6A profiling by methylated RNA-immunoprecipitation sequencing revealed m6A peaks near the stop codon in mRNAs of multiple oncogenes including MAF and CCND1. Cross-linking immunoprecipitation showed that METTL3 bound to the m6A peak within MAF and CCND1 mRNA. Depletion of METTL3 by shRNA had little effect on global mRNA levels, but specifically reduced protein levels of c-Maf and Cyclin D1. Moreover, downregulation of METTL3 in several MM cell lines results in cell cycle arrest and apoptosis. Together, these results describe a role for METTL3 in promoting translation of a subset of oncogenes in MM and identify this enzyme as a potential therapeutic target for multiple myeloma. Disclosures No relevant conflicts of interest to declare.

scholarly journals Downregulated KLF2 in PV and ET May Induce Prothrombotic Gene Expression

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 13-14
Author(s):  
Jihyun Song ◽  
Jahnavi Gollamudi ◽  
Soo Jin Kim ◽  
Radhika Gangaraju ◽  
Tsewang Tashi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Tissue Factor ◽  
Research Funding ◽  
Interleukin 1 ◽  
Negative Regulator ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Inherited Disorders ◽  
History Of

Polycythemia vera (PV) and essential thrombocythemia (ET) patients have a higher risk of arterial and venous thrombosis than healthy individuals; thromboses are their principal cause of morbidity and mortality. We reported increased transcription of prothrombotic and inflammatory genes in granulocytes and platelets of PV and ET. There were differences in the expression of prothrombotic genes between platelets and granulocytes, suggesting that these cells have cell-specific contributions to thrombosis in PV and ET. Some of these prothrombotic genes are regulated by hypoxia inducible factors (HIFs) (PMID: 32203583). However, the molecular mechanism of thrombosis in PV and ET remains unknown. KLF2 (Kruppel like factor 2) is a transcription factor regulating primitive erythropoiesis and inflammation. Knockdown of KLF2 in cultured endothelial cells increases prothrombotic gene expression and reduces blood clotting time and flow rates (PMID: 15718498). Targeted deletion of KLF2 in neutrophils increases thrombosis by inducing the expression and activity of tissue factor (Blood, 2018, 132:75). To study the role of KLF2 in PV and ET thrombosis, we measured KLF2 mRNA in granulocytes from 53 PV and ET patients (25 with a history of thrombosis) and in platelets from 40 patients (21 with a history of thrombosis). We also measured KLF2 mRNA in granulocytes from 38 controls and platelets from 18 controls. Althrough the role of KLF2 in thrombosis has been studied in neutrophils, we also tested KLF2 mRNA in platelets since we previously observed a different pattern of expression of prothrombotic genes between granulocytes and platelets in PV and ET. We found lower KLF2 mRNA in both granulocytes and platelets of PV and ET patients compared to the controls (Fig. 1A and B). Compared to patients without thrombosis, those with thrombosis had lower KLF2 mRNA in platelets but not granulocytes. KLF2 mRNA in these cells correlated inversely with JAK2V617F allele burden in granulocytes (Fig. 1C and D). We then measured mRNA of prothrombotic genes: F3 (tissue factor), SELP (P-selectin), IRAK1 (interleukin 1 receptor associated kinase 1), IL1RAP (interleukin 1 receptor accessory protein), VEGFA (vascular endothelial growth factor-A), THSB1 (thrombospondin 1), SERPINE1 (encoding plasminogen activator inhibitor 1 [PAI-1]). The mRNA levels of these prothrombotic genes correlated inversely with KLF2 mRNA in platelets while SELP and THSB1 transcripts correlated inversely with KLF2 mRNA in granulocytes. KLF2 and HIFs are reported to interact (PMID: 19491109, PMID: 21565532). In order to elucidate the regulatory machanism of KLF2 in thrombosis, we measured KLF2 mRNA in patients with two inherited disorders of hypoxia sensing characterized by thrombosis: 1) Chuvash erythrocytosis (CE) due to homozygous mutation of VHLR200W (13 patients) and 2) erythrocytosis due to gain-of-function mutation of HIF-2a (two patients with HIF2AM535V and two patients HIF2AE548K). KLF2 mRNA levels did not differ in granulocytes and platelets between these patients and controls. However, two CE patients and two patients with HIF2AM535V with a history of thrombosis had lower KLF2 mRNA levels compared to patients without thrombosis (ASH this meeting, 2020 Song J). In conclusion, we report here that KLF2 transcripts are down regulated in both granulocytes and platelets from PV and ET patients and they correlate inversely with the transcripts of prothrombotic genes and JAK2V617F allelic burden, suggesting that KLF2 might be a negative regulator of thrombotic gene expression in PV and ET. Here we did not detect any changes of KLF2 transcripts in congential disorders with elevated HIFs. However, two CE patients and 2 patients with HIF2AM535V with thrombosis had less KLF2 expression compared to those without thrombosis. These results suggest that, by inference from findings in congenital disorders with elevated HIFs, KLF2 in PV and ET granulocytes and platelets may be regulated in a HIF-independent manner but that thrombosis may be regulated in a HIF-dependent manner. Thus, KLF2 may be a novel therapeutic target to prevent thrombosis in PV and ET, but confirmation by further studies is needed. The upstream regulation of KLF2 in PV and ET granulocytes and platelets needs to be elucidated. *PT &JTP contributed equally Disclosures Gangaraju: Sanofi Genzyme, Consultant for Cold Agglutinin Disease: Consultancy. Gordeuk:CSL Behring: Consultancy, Research Funding; Global Blood Therapeutics: Consultancy, Research Funding; Imara: Research Funding; Ironwood: Research Funding; Novartis: Consultancy.

scholarly journals CRP Is Transported by Monocytes and Monocyte-Derived Exosomes in the Blood of Patients with Coronary Artery Disease

Biomedicines ◽  
2020 ◽  
Vol 8 (10) ◽  
pp. 435
Author(s):  
Ivan Melnikov ◽  
Sergey Kozlov ◽  
Olga Saburova ◽  
Ekaterina Zubkova ◽  
Olga Guseva ◽  
...  
Keyword(s):  
Coronary Artery Disease ◽  
Coronary Artery ◽  
Healthy Volunteers ◽  
Blood Cells ◽  
Messenger Rna ◽  
Blood Monocytes ◽  
Gm Csf ◽  
Reactive Protein ◽  
Artery Disease ◽  
Macrophage Colony

The objective of this work was to study the ability of blood cells and their microparticles to transport monomeric and pentameric forms of C-reactive protein (mCRP and pCRP) in the blood of patients with coronary artery disease (CAD). Blood was obtained from 14 patients with CAD 46 ± 13 years old and 8 healthy volunteers 49 ± 13.6 years old. Blood cells and microparticles with mCRP and pCRP on their surface were detected by flow cytometry. Messenger RNA (mRNA) of CRP was extracted from peripheral blood monocytes stimulated with lipopolysaccharide (LPS) and granulocyte-macrophage colony-stimulating factor (GM-CSF). mRNA of CRP in monocytes was detected with PCR. Monocytes were predominantly pCRP-positive (92.9 ± 6.8%). mCRP was present on 22.0 ± 9.6% of monocyte-derived exosomes. mCRP-positive leukocyte-derived microparticle counts were significantly higher (8764 ± 2876/µL) in the blood of patients with CAD than in healthy volunteers (1472 ± 307/µL). LPS and GM-CSF stimulated monocytes expressed CRP mRNA transcripts levels (0.79 ± 0.73-fold), slightly lower relative to unstimulated hepatocytes of the HepG2 cell line (1.0 ± 0.6-fold), but still detectable. The ability of monocytes to transport pCRP in blood flow, and monocyte-derived exosomes to transmit mCRP, may contribute to the maintenance of chronic inflammation in CAD.

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