scholarly journals C-reactive protein induces human peripheral blood monocytes to synthesize tissue factor

Blood ◽  
1993 ◽  
Vol 82 (2) ◽  
pp. 513-520 ◽  
Author(s):  
J Cermak ◽  
NS Key ◽  
RR Bach ◽  
J Balla ◽  
HS Jacob ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
De Novo ◽  
Human Peripheral Blood ◽  
Umbilical Vein ◽  
Blood Monocytes ◽  
C Reactive Protein ◽  
Dot Blot ◽  
Reactive Protein

The acute inflammatory response is frequently accompanied by serious thrombotic events. We show that C-reactive protein (CRP), an acute- phase reactant that markedly increases its serum concentration in response to inflammatory stimuli, induced monocytes to express tissue factor (TF), a potent procoagulant. Purified human CRP in concentrations commonly achieved in vivo during inflammation (10 to 100 micrograms/mL) induced a 75-fold increase in TF procoagulant activity (PCA) of human peripheral blood mononuclear cells (PBM), with a parallel increase in TF antigen levels. CRP-induced PCA was completely blocked by a monoclonal antibody against human TF but not by irrelevant murine IgG. Dot blot analysis showed a significant increase of TF mRNA after 4 hours of incubation with CRP, followed by a peak of PCA within 6 and 8 hours. Actinomycin D and cycloheximide blocked CRP-stimulated PCA, suggesting that de novo TF protein synthesis was required. Endotoxin (LPS) contamination of CRP was excluded as the mediator of TF synthesis because: (1) CRP was Limulus assay negative; (2) induction of TF PCA by CRP was not blocked by Polymyxin B, in contrast to LPS- induced PCA; (3) antihuman CRP IgG inhibited CRP-induced PCA, but not LPS-induced PCA; (4) CRP was able to stimulate TF production in LPS- pretreated PBM refractory to additional LPS stimulation; and, (5) unlike LPS, CRP was incapable of inducing TF in human umbilical vein endothelial cells. We suggest that CRP-mediated TF production in monocytes may contribute to the development of disseminated intravascular coagulation and thrombosis in inflammatory states.

scholarly journals C-reactive protein induces human peripheral blood monocytes to synthesize tissue factor

Blood ◽  
1993 ◽  
Vol 82 (2) ◽  
pp. 513-520 ◽  
Author(s):  
J Cermak ◽  
NS Key ◽  
RR Bach ◽  
J Balla ◽  
HS Jacob ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
De Novo ◽  
Human Peripheral Blood ◽  
Umbilical Vein ◽  
Blood Monocytes ◽  
C Reactive Protein ◽  
Dot Blot ◽  
Reactive Protein

Abstract The acute inflammatory response is frequently accompanied by serious thrombotic events. We show that C-reactive protein (CRP), an acute- phase reactant that markedly increases its serum concentration in response to inflammatory stimuli, induced monocytes to express tissue factor (TF), a potent procoagulant. Purified human CRP in concentrations commonly achieved in vivo during inflammation (10 to 100 micrograms/mL) induced a 75-fold increase in TF procoagulant activity (PCA) of human peripheral blood mononuclear cells (PBM), with a parallel increase in TF antigen levels. CRP-induced PCA was completely blocked by a monoclonal antibody against human TF but not by irrelevant murine IgG. Dot blot analysis showed a significant increase of TF mRNA after 4 hours of incubation with CRP, followed by a peak of PCA within 6 and 8 hours. Actinomycin D and cycloheximide blocked CRP-stimulated PCA, suggesting that de novo TF protein synthesis was required. Endotoxin (LPS) contamination of CRP was excluded as the mediator of TF synthesis because: (1) CRP was Limulus assay negative; (2) induction of TF PCA by CRP was not blocked by Polymyxin B, in contrast to LPS- induced PCA; (3) antihuman CRP IgG inhibited CRP-induced PCA, but not LPS-induced PCA; (4) CRP was able to stimulate TF production in LPS- pretreated PBM refractory to additional LPS stimulation; and, (5) unlike LPS, CRP was incapable of inducing TF in human umbilical vein endothelial cells. We suggest that CRP-mediated TF production in monocytes may contribute to the development of disseminated intravascular coagulation and thrombosis in inflammatory states.

scholarly journals Early-Stage Identification and Avoidance of Antisense Oligonucleotides Causing Species-Specific Inflammatory Responses in Human Volunteers Peripheral Blood Mononuclear Cells

2021 ◽  
Author(s):  
Sebastien AL Burel ◽  
Brenda F Baker ◽  
T. Jesse Kwoh ◽  
Suzanne Paz ◽  
Husam Younis ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Inflammatory Responses ◽  
Early Stage ◽  
Human Peripheral Blood ◽  
C Reactive Protein ◽  
Healthy Human ◽  
Peripheral Blood Mononuclear ◽  
Reactive Protein ◽  
Human Volunteers

A human peripheral blood mononuclear cell (PBMC)-based assay was developed to identify antisense oligonucleotide (ASO) with the potential to activate a cellular innate immune response outside of an acceptable level. The development of this assay was initiated when ISIS 353512 targeting the mRNA for human C-reactive protein was tested in a phase I clinical trial in which healthy human volunteers unexpectedly experienced increases in interleukin-6 (IL-6) and C-reactive protein. This level of immune stimulation was not anticipated following rodent and non-human primate safety studies in which no evidence of exaggerated proinflammatory effects were observed. The IL-6 increase induced by ISIS 353512 was caused by activation of B-cells. The IL-6 induction was inhibited by chloroquine pretreatment of the PBMC and the nature of the ASOs suggested that the response is mediated by a toll like receptor, in all likelihood TLR9. While assessing the inter PBMC donor variability, two class of responders to ISIS 353512 were identified (discriminator and non-discriminators). The discriminator source of PBMC was shown to produce low level of IL-6 after 24 hours in culture in absence of ASO treatment. The PBMC assay using discriminator donors was shown to be reproducible allowing to assess reliably the immune potential of ASOs via comparison to known benchmark ASO controls that were previously shown to be either safe or inflammatory in clinical trials.

The Effect of C-Reactive Protein (CRP) on Respiratory Burst of Human Peripheral Blood Phagocytes

1995 ◽  
Vol 8 (2) ◽  
pp. 95-102
Author(s):  
J. Cofta ◽  
J.K. Lacki ◽  
S.H. Mackiewicz ◽  
K.E. Wiktorowicz
Keyword(s):  
Respiratory Burst ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Low Doses ◽  
Blood Monocytes ◽  
C Reactive Protein ◽  
Free Oxygen ◽  
Peripheral Blood Monocytes ◽  
Reactive Protein ◽  
Enhanced Chemiluminescence

The effect of C-reactive protein (CRP) on the oxidative response of human peripheral blood monocytes and granulocytes was investigated. The respiratory burst of phagocytes induced by phorbol-myristate-13-acetate (PMA), phytohaemagglutinin (PHA) or opsonized zymosan (OZ) was measured in luminol-enhanced chemiluminescence. The effect of CRP on PMA-induced monocyte chemiluminescence (CL) depended both on CRP concentration and incubation time. A short incubation of cells with CRP (15–30 min.) enhanced the oxidative burst. Preincubation of cells for 1h (or longer) with low doses of CRP (about 2 μg/ml) increased, while with higher (>10 μg/ml) inhibited PMA-stimulated chemiluminescence. CRP reduced also PHA or OZ-induced monocyte respiratory response. CRP diminished PMA, PHA, and OZ-induced granulocyte chemiluminescence, except the response to PHA in the presence of low doses CRP (about 5 μg/ml). The action of CRP on phagocytes probably involves activation of some intracellular mechanisms. During immune response, CRP could protect tissues against damage by excess of free oxygen and its biological active derivatives.

scholarly journals Peroxynitrite reduces endotoxin-induced tissue factor expression in human peripheral blood monocytes

Critical Care ◽  
10.1186/cc29 ◽  
1997 ◽  
Vol 1 (Suppl 1) ◽  
pp. P023
Author(s):  
M Gerlach ◽  
D Keh ◽  
S Spielmann ◽  
T Kerner ◽  
R Peter ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Tissue Factor Expression ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Factor Expression

INHIBITION OF ENDOTHELIAL CELL PROLIFERATION BY NORMAL HUMAN MONOCYTES

1987 ◽  
Author(s):  
F Liote ◽  
M P Wautier ◽  
E Savariau ◽  
H Setiadi ◽  
J L Wautier
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Umbilical Vein ◽  
Endothelial Cell Proliferation ◽  
Thymidine Uptake ◽  
Blood Monocytes ◽  
Fibronectin Receptor

Human peripheral blood monocytes and macrophages possess factors which are capable of inhibiting or stimulating endothelial cell proliferation. We have further explored if such activity is due to cytotoxic effects of monocytes. Normal mononuclear cells were isolated first by density gradient. Monocytes were then purified by three different techniques: 1) counter centrifugation elutriation (CCE) (Beckman) 2) selective adhesion to gelatin-plasma (GPI) 3) selective adhesion to fibronectin (Fn). Cytotoxicity was estimated by counting the release of 51cr used to label the human umbilical vein endothelial cells (HUVE) prior to the addition of monocytes. Whilst [3H] thymidine incorporation by HUVE permitted us to measure the effect of monocytes on the growth of the endothelial cells. Monocytes were incubated with HUVE (12×103) for 24 to 36h at various concentrations '(1.5-12×103). No cytotoxic effect could be demonstrated but an inhibition of [3h] thymidine uptake was observed and was dependent upon monocytes concentration. Monocytes isolated on GP1 exhibited a significantly higher inhibitory effect (p<0.05) compared to those purified on Fn or by CCE.(GP1: 85±6%, Fn:58±6%, CCE:67±5%). These results indicated t*hat normal monocytes can inhibit endothelial cell proliferation. This activity appeared to be higher when monocytes were isolated on GP1 which suggest that the adhesion on this surface could stimulate monocytes not only by its fibronectin receptor. This inhibitory activity of monocyte on endothelial cells proliferation could be different in patients with vascular disorders.

C-Reactive Protein Binds to FcγRII and Impairs the Differentiation and Function of Dendritic Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1276-1276
Author(s):  
Jianfei Qian ◽  
Jing Yang ◽  
Siqing Wang ◽  
Liang Zhang ◽  
Sungyoul Hong ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Soluble Antigen ◽  
Dependent Manner ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Tnf Α ◽  
And Function

Abstract Human C-reactive protein (CRP) is an acute-phase protein, and elevated levels of CRP are present in patients with infections, inflammatory diseases, necrosis such as myocardial infarction, or malignancies including multiple myeloma (MM), lymphoma, and carcinoma. CRP has many biological functions and is involved in host defense, regulation of inflammation, and modulation of autoimmune diseases. Although our current understanding of CRP interaction with complement and Fcγ receptors (FcγR) has elucidated a regulatory role of CRP in these disease situations, it is not clear whether CRP affects the function of immune cells such as dendritic cells (DCs). In this study, we investigated the effect of CRP on DC differentiation, maturation, and function. CD14+ monocytes isolated from human peripheral blood mononuclear cells were cultured in RPMI-1640 medium supplemented with GM-CSF and IL-4 for 5 days to generate immature DCs (imDCs), and were further treated with IL-1β and TNF-α for 2 additional days to produce mature DCs (mDCs). CRP (5–100 μg/mL) was added to the cultures during DC differentiation (on days 0 and 3) or maturation (on day 5). The presence of CRP in cultures reduced imDC cell yields in a dose-dependent manner. Significantly lower cell yields were detected in cultures with 5 to 10 μg/mL CRP. Compared with untreated controls, CRP treatment (10 μg/mL) led to inhibited surface expression of DC-related molecules HLA-ABC, CD1a, CD40, and CD54; increased secretion of IL-6, IL-8, and IL-10; reduced production of TGF-β by imDCs; and decreased secretion of IL-12 by mDCs. Furthermore, the function of CRP-treated DCs was also impaired, evident by the markedly decreased ability of imDCs to phagocytose apoptotic cells and to uptake and present soluble antigen to antigen-specific T cells. Compared with untreated controls, CRP-treated mDCs had reduced capacity at activating allospecific T cells, which consequently secreted significantly lower amounts of IFN-γ, IL-2, and TNF-α compared with T cells activated by normal mDCs. Western blot analysis showed that CRP treatment led to inhibited phosphorylation of ERK and p38 MAPKs, and inhibited NFκB activity in the differentiating cells. Monocytes and DCs all express FcγRI (CD64), FcγRII (CD32), and FcγRIII (CD16), and the expression of FcγRII, but not FcγRI and FcγRIII, were upregulated on CRP-treated DCs. The detrimental effects of CRP on DCs were abrogated by blocking antibody against CRP and by antibody against FcγRII, but not against FcγRI or FcγRIII. These results indicate that CRP affected DC differentiation via binding to cell surface FcγRII. Taken together, this study demonstrates for the first time that CRP at high concentrations has detrimental effects on in vitro differentiation and function of DCs. Further studies will be needed to examine the clinical and biological relevance of this observation.

The Blood Monocyte - TNF - Endothelial Axis in Activation of Endothelial Tissue Factor in the Transgenic Sickle Mouse: Possible Relevance of Etanercept to Sickle Coagulopathy

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1048-1048
Author(s):  
Anna Solovey ◽  
Liming Milbauer Chang ◽  
Fuad Abdulla ◽  
Rahn Kollander ◽  
Paul H Marker ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Conflicts Of Interest ◽  
Pulmonary Veins ◽  
Ambient Air ◽  
Blood Monocytes ◽  
Extended Treatment ◽  
Pre Treatment ◽  
Endothelial Tissue

Abstract Abstract 1048 Peripheral blood monocytes can be pathogenic participants in vascular disease, and they are abnormally activated in sickle patients. As shown earlier, sickle mice have abnormally increased expression of endothelial tissue factor (eTF) in pulmonary veins (PV), expressed as % of PV positive for eTF. For HbS-BERK (eTF 36%, vs 9% for HbA-BERK controls) and S+SAntilles (eTF 28%, vs 7% for C57BL6 controls), abnormal eTF expression is chronic. For NY1DD, however, eTF switches from 7% at ambient air to 23% after exposure to hypoxia/reoxygenation (post-H/R). Having now examined roles of transcription factors Egr-1 and NFkB(p50), we find that H/R triggered expression of eTF in post-H/R NY1DD is: [a] dependent upon both Egr-1 and NFkB(p50) within peripheral blood mononuclear cells (PBMC), and dependent upon Egr-1 butnot NFkB(p50) in vessel wall endothelium. To examine the roles of PBMC and their product TNF in activating eTF expression we used three strategies. [A] First, transfusion (Tx) of PBMC from donor mice (DM) induced eTF in recipient mice (RM) as follows. At-air NY1DD RM exhibited eTF of 7% from Tx of at-air NY1DD DM PBMC, and eTF of 19% from Tx of post-H/R NY1DD DM PBMC. C57BL6 RM exhibited eTF of 10% from Tx of C57BL6 DM PBMC, 31% from Tx of S+SAntilles DM PBMC, and only 15% from Tx of PBMC from S+SAntilles DM also having NFkB(p50)−/−. HbA-BERK RM exhibited eTF of 9% from Tx of HbA-BERK DM PBMC, and 36% from Tx of HbS-BERK DM PBMC. [B] Second, we examined effect of TNF+LT blocker etanercept on PBMC in transfusion experiments. As expected, HbA-BERK RM exhibited eTF 36% from Tx of HbS-BERK PBMC obtained from etanercept-treated (3 mg/kg × 2) DM, and 38% from Tx of HbS-BERK PBMC obtained from DM treated with inactive control TNF-R-scrambled-peptide. On the other hand, HbA-BERK RM pre-treated with etanercept (same dose) vs control TNF-R-scrambled-peptide exhibited eTF of 11% and 33%, respectively from Tx of HbS-BERK DM PBMC. [C] Third, direct TNF blocking pre-treatment of NY1DD mice prevented the eTF increase from subsequent H/R (to 23% for the no pre-treatment controls): eTF of 5% (P=.000013) for pre-treatment with high-etanercept (10 mg/kg × 2), 12% (P=.00059) for pre-treatment with low-etanercept (3 mg/kg × 2), 19% (P=.053) for pre-treatment with IL1-blocker anakinra (10 mg/kg ×2), and 7% (P=.0016) for pretreatment withTNF-specific blocker infliximab (10 mg/kg ×2). For HbS-BERK mice having pre-existing eTF of 36%, extended treatment with etanercept (3 mg/kg, twice weekly × 3) reduced eTF to 27% (P=.026) while those receiving scrambled-peptide remained at 34%. Higher or longer etanercept may be more effective. Other murine treatments have shown us reversal of pre-existing eTF requires extended treatment. Since regulation of TF expression in endothelial cells and blood monocytes is similar, it is likely that these eTF effects would be paralleled by similar effects on monocyte TF expression. Thus, these results suggest that TNF blocking strategies could be helpful in mitigating effects of coagulation activation in the sickle context. Disclosures: No relevant conflicts of interest to declare.

Sign in / Sign up

Export Citation Format

Share Document