Increased Concentrations of Soluble CD40 Ligand, RANTES and GRO-α in Preeclampsia – Possible Role of Platelet Activation

SummaryActivated platelets may release inflammatory mediators that activate leukocytes and trigger inflammatory reactions in endothelial cells. We examined the concentrations of soluble CD40 ligand (sCD40L) and the chemokines RANTES and GRO-α in platelet-free plasma (PFP), and unstimulated and SFLLRN-stimulated platelet-rich plasma (PRP), as well as in platelet pellets before stimulation using enzyme immunoassays. Nineteen women with normal and twenty-one with preeclamptic pregnancies were studied, and several differences between these two groups of pregnancies were revealed (1). Women with preeclampsia had significantly increased concentrations of sCD40L and GRO-α in PFP (2). Platelets from these patients spontaneously released larger quantities of CD40L and RANTES ex vivo (3). When further activated ex vivo by SFLLRN, platelets from preeclamptic women released lower amounts per platelet of CD40L, RANTES and GRO-α (4). The platelet pellets in preeclamptic women contained decreased amounts of CD40L, RANTES and GRO-α per platelet. Our findings suggest enhanced platelet activation in vivo during preeclampsia resulting in increased release of inflammatory mediators, possibly contributing to inflammation, leukocyte activation and endothelial dysfunction in this disorder.

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Internalization of IgG-Coated Targets Results in Activation and Secretion of Soluble CD40 Ligand and RANTES by Human Platelets

Clinical and Vaccine Immunology ◽

10.1128/cvi.00296-10 ◽

2010 ◽

Vol 18 (2) ◽

pp. 210-216 ◽

Cited By ~ 24

Author(s):

Adam J. Antczak ◽

Joshua A. Vieth ◽

Navinderjit Singh ◽

Randall G. Worth

Keyword(s):

Platelet Activation ◽

Cytokine Production ◽

Cd40 Ligand ◽

Human Platelets ◽

Cytochalasin D ◽

Soluble Cd40 Ligand ◽

Actin Remodeling ◽

Inflammatory Reactions ◽

Coated Particles ◽

Immunological Reactions

ABSTRACTPlatelets are crucial elements for maintenance of hemostasis. Other functions attributable to platelets are now being appreciated, such as their role in inflammatory reactions and host defense. Platelets have been reported to bind immunological stimuli like IgG complexes, and for nearly 50 years it has been speculated that platelets may participate in immunological reactions. Platelets have been reported to bind and internalize various substances, similar to other leukocytes, such as macrophages and dendritic cells. In the present study, we tested the hypothesis that human platelets can bind and internalize IgG-coated particles, similar to leukocytes. To this end, we observed that interaction with IgG-coated beads resulted in platelet activation (as measured by CD62P expression), internalization of targets, and significant soluble CD40 ligand (sCD40L) and RANTES (regulated uponactivation,normalTcellexpresses andsecreted) secretion. Blocking FcγRIIA with monoclonal antibody (MAb) IV.3 or inhibiting actin remodeling with cytochalasin D inhibited platelet activation, internalization, and cytokine production. These data suggest that platelets are capable of mediating internalization of IgG-coated particles, resulting in platelet activation and release of both sCD40L and RANTES.

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Platelet-specific markers are associated with monocyte-platelet aggregate formation and thrombin generation potential in advanced atherosclerosis

Thrombosis and Haemostasis ◽

10.1160/th15-07-0598 ◽

2016 ◽

Vol 115 (03) ◽

pp. 615-621 ◽

Cited By ~ 17

Author(s):

Cihan Ay ◽

Julia Riedl ◽

Christoph W. Kopp ◽

Beate Eichelberger ◽

Renate Koppensteiner ◽

...

Keyword(s):

Platelet Activation ◽

Thrombin Generation ◽

Thrombus Formation ◽

Cd40 Ligand ◽

Aggregate Formation ◽

Soluble Cd40 Ligand ◽

Platelet Factor ◽

Platelet Aggregate ◽

Angioplasty And Stenting

SummaryPlatelet activation and thrombin generation are crucial steps in primary and secondary haemostasis. However, both also play major roles in intravascular thrombus formation and therefore in the development of adverse cardiovascular events. In the current study, we first sought to investigate the associations of the platelet biomarkers platelet factor (PF)-4, thrombospondin (TSP)-1, soluble CD40 ligand (sCD40L), and soluble P-selectin (sP-selectin) with each other and with monocyte-platelet aggregate (MPA) formation in 316 patients undergoing angioplasty and stenting. To better understand the interplay between platelet activation and thrombin generation, we subsequently investigated the associations of the platelet biomarkers with thrombin generation potential. The mostly platelet-specific markers PF-4, TSP-1 and sCD40L correlated strongly with each other (all p < 0.001), and the best correlation was observed between PF-4 and TSP-1 (r=0.91). In contrast, sP-selectin, which derives from platelets and endothelial cells, correlated rather poorly with TSP-1 (r=0.12, p=0.04), and did not correlate with PF-4 and sCD40L. While PF-4, TSP-1 and sP-selectin correlated significantly with in vivo MPA formation (all p < 0.001), no such association was found between sCD40L and MPA formation. PF-4, TSP-1 and sCD40L correlated strongly with peak thrombin generation (all p < 0.001) with the best correlation between PF-4 and peak thrombin generation (r=0.55), whereas sP-selectin did not correlate with peak thrombin generation. Likewise, PF-4, TSP-1 and sCD40L correlated significantly with the area under the thrombin generation curve (AUC; all p< 0.01), whereas sP-selectin did not correlate with the AUC. In conclusion, platelet-specific markers are associated with MPA formation and thrombin generation potential in patients with advanced atherosclerosis.

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Possible Role Of Circulating Adenine Nucleotides and Platelet Activation In The Pathogenesis Of Sickle Cell Disease: Observations In a Murine Model

Blood ◽

10.1182/blood.v122.21.2212.2212 ◽

2013 ◽

Vol 122 (21) ◽

pp. 2212-2212

Author(s):

Kousaku Ohno ◽

Hisako Tanaka ◽

Naozumi Samata ◽

Tomizawa Atsuyuki ◽

Makoto Mizuno ◽

...

Keyword(s):

Platelet Activation ◽

Matched Control ◽

Ex Vivo ◽

Adenine Nucleotides ◽

Significant Inhibition ◽

Cell Disease ◽

Basal Expression ◽

Old Mice

Abstract Vaso-occlusive crisis (VOC) is common in patients with sickle cell disease (SCD). Although hemolysis of erythrocytes has been associated with VOC, the underlying mechanism remains unclear. In the present study, we conducted pathophysiological analyses in Berkeley SCD mice to examine the possible role of adenine nucleotides and platelet activation in SCD. Berkeley SCD mice are genetically modified mice whose murine α-globin and β-globin genes have been deleted, and replaced with human α-, γ-, and δ-globins and sickle β-globin. Control mice were heterozygous littermates expressing normal β-globin. Histological analyses revealed congestion of blood vessels in the liver vasculature and renal medulla of 7-week-old SCD mice associated with aggregated sickle-shaped erythrocytes. Further analysis revealed severe hemolytic anemia in these 7-week-old SCD mice. Plasma adenine nucleotide measurements using LC-MS/MS showed that the concentrations of ADP in 7-week-old SCD mice were significantly higher (∼2.7-fold increase) compared with age-matched control mice. These findings suggest that the intravascular hemolysis of sickled erythrocytes and/or congestion in various organs results in the elevation of circulating concentrations of ADP in this SCD model. Platelet function was assessed by flow-cytometric analysis of platelet-surface activated GPIIbIIIa and P-selectin. These biomarkers of platelet activation were determined in vehicle-treated control mice, vehicle-treated SCD mice, and prasugrel (1 mg/kg/day)-treated SCD mice. Vehicle or prasugrel were orally administered to 7-week-old mice for 1 week and 8 weeks. The expression of activated GPIIbIIIa on platelets was assessed with or without 20 μM ADP stimulation. The expression of platelet P-selectin was assessed with or without 300 μM PAR4 TRAP. These platelet function analyses showed that circulating platelets in SCD mice were activated compared to control mice; the basal expression of activated GPIIbIIIa on platelets in 8- and 15-week-old SCD mice and P-selectin on platelets in 15-week-old SCD mice were increased significantly compared with those in control mice. In vitro stimulation with ADP and PAR4 TRAP increased the expression of activated GPIIbIIIa and P-selectin on platelets in SCD mice. Of note, agonist-stimulated activation was enhanced with age. In 8-week-old mice, the extent of agonist-induced activation of GPIIbIIIa and P-selectin in SCD mice was equivalent to that in control mice; however, in 15-week-old mice, the extent of agonist-induced activation of platelet GPIIbIIIa and P-selectin in SCD mice was significantly higher compared with that in age matched control mice. Prasugrel treatment for 1 week and 8 weeks did not result in significant inhibition of basal expression of activated GPIIbIIIa or P-selectin on circulating platelets in SCD mice, suggesting P2Y12 independent activation of circulating platelets. In contrast, prasugrel treatment for 1 week and 8 weeks did result in highly significant inhibition of ADP-induced GPIIbIIIa activation on SCD mouse platelets. PAR4 TRAP-induced P-selectin expression on SCD mouse platelets was also inhibited by prasugrel. In conclusion, in a murine model of SCD, the present study found significant elevation of circulating ADP possibly resulting from the in vivo hemolysis of sickled erythrocytes. However, P2Y12 inhibition by prasugrel did not appear to reduce the activation of circulating platelets in this SCD model. In contrast, ex vivo stimulation of SCD mouse platelets by the ex vivo addition of ADP and PAR4 TRAP was effectively inhibited by prasugrel administration. The ex vivo addition of ADP may mimic increased exposure to ADP in vivo during VOC thus suggesting a role for prasugrel in the treatment of SCD. Disclosures: Ohno: Daiichi Sankyo Co., Ltd.: Employment. Tanaka:Daiichi Sankyo RD Novare Co., Ltd.: Employment. Samata:Daiichi Sankyo RD Novare Co., Ltd.: Employment. Atsuyuki:Daiichi Sankyo Co., Ltd.: Employment. Mizuno:Daiichi Sankyo Co., Ltd.: Employment. Jakubowski:Eli Lilly and Company: Employment. Sugidachi:Daiichi Sankyo Co., Ltd.: Employment.

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Increased Plasma Concentrations of Soluble CD40 Ligand in Acute Coronary Syndrome Depend on in Vitro Platelet Activation

Clinical Chemistry ◽

10.1373/clinchem.2007.085332 ◽

2007 ◽

Vol 53 (7) ◽

pp. 1231-1234 ◽

Cited By ~ 33

Author(s):

Boris T Ivandic ◽

Eberhard Spanuth ◽

Detlef Haase ◽

Heiko-Gundmar Lestin ◽

Hugo A Katus

Keyword(s):

Acute Coronary Syndrome ◽

Platelet Activation ◽

Room Temperature ◽

Cd40 Ligand ◽

Soluble Cd40 Ligand ◽

Platelet Factor ◽

Coronary Syndrome ◽

In Vitro Platelet Activation

Abstract Background: Soluble CD40 ligand (sCD40L) was suggested as a novel biomarker of cardiovascular risk. We examined the effect of preanalytical variation on the measurement of sCD40L concentration. Methods: From healthy control individuals (n = 20) and patients with acute coronary syndrome (ACS) (n = 20) or sepsis (n = 20), we obtained blood drawn into 5 tubes containing citrate or a mixture of citrate, theophylline, adenosine, and dipyridamole (CTAD). The tubes were incubated for 30 min at room temperature or 0 °C before a single or double centrifugation (15 min, 2500g) at room temperature or 4 °C, respectively. sCD40L, β-thromboglobulin (βTG), and platelet factor 4 (PF4) concentrations were measured using immunoassays. Results: Concentrations of sCD40L were very low in all CTAD and citrated samples maintained at 0 °C (median ≤0.076 μg/L). Although increased βTG and PF4 confirmed disease-related in vivo platelet activation, sCD40L was not higher in patients than in controls. In contrast, if the samples were processed at room temperature, sCD40L was significantly higher in ACS patients than in controls (P <0.02 in CTAD and citrated plasma at room temperature). Moreover, the βTG:PF4 ratio decreased in patient but not control CTAD samples, suggesting a greater susceptibility of patient platelets to in vitro activation. Conclusions: Increased sCD40L concentrations resulted from in vitro platelet activation during sample preparation. Disease-related in vivo activation did not contribute to sCD40L concentrations in plasma. Therefore, published studies of sCD40L demand cautious interpretation, because their preanalytical conditions were not standardized.

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Abstract 3995: Comparison of the Effects of Prasugrel and High Dose Clopidogrel on In Vivo and In Vitro Platelet Activation: Results from PRINCIPLE-TIMI 44

Circulation ◽

10.1161/circ.118.suppl_18.s_814-a ◽

2008 ◽

Vol 118 (suppl_18) ◽

Author(s):

Andrew L Frelinger ◽

Alan D Michelson ◽

Eugene Braunwald ◽

Dietmar Trenk ◽

Franz-Josef Neumann ◽

...

Keyword(s):

Platelet Activation ◽

Thrombus Formation ◽

Inflammatory Marker ◽

Randomized Study ◽

Cd40 Ligand ◽

High Dose ◽

List Type ◽

Soluble Cd40 Ligand

In PRINCIPLE-TIMI 44, a randomized study in 201 patients undergoing cardiac catheterization for planned PCI, we reported that prasugrel resulted in greater inhibition of in vitro ADP-stimulated platelet aggregation than clopidogrel 600 mg loading dose (LD) and 150 mg maintenance dose (MD). Here we compare the effects of prasugrel and clopidogrel on in vivo and in vitro platelet activation. As prespecified end points of PRINCIPLE-TIMI 44, we measured circulating platelet-monocyte aggregates (PMA), circulating platelet-neutrophil aggregates (PNA), circulating P-selectin-positive platelets (PSPP), plasma soluble CD40 ligand (sCD40L) and (because activated platelets induce its release from leukocytes) myeloperoxidase (MPO). We also evaluated in vitro ADP-stimulated PMA, PNA and PSPP. At 6 h post LD, clopidogrel and prasugrel resulted in similar reductions in circulating PMA (Fig. A ), PNA and PSPP. In vitro ADP-stimulated PMA (Fig. B ), PNA and PSPP were inhibited to a greater degree by prasugrel than clopidogrel during the LD and MD phases. At 6 h post LD, clopidogrel- but not prasugrel-treated patients had elevated plasma MPO compared to pre-treatment (Fig. C ). Plasma sCD40L was unaffected by time or treatment. In this randomized study: Prasugrel, like clopidogrel, inhibited in vivo platelet activation. Prasugrel, to a greater degree than clopidogrel, inhibited ADP-stimulated PMA, PNA and PSPP – which may be mechanistically important, given the high local concentrations of ADP at sites of thrombus formation. Prasugrel, unlike clopidogrel, prevented the post-PCI-induced increase in the inflammatory marker MPO.

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Role of Red Blood Cells in the Early Stages of Platelet Activation by Collagen

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661686 ◽

1986 ◽

Vol 56 (03) ◽

pp. 376-381 ◽

Cited By ~ 27

Author(s):

M Teresa Santos ◽

Juana Vallés ◽

Justo Aznar ◽

J Luis Pérez-Requejo

Keyword(s):

Red Blood Cells ◽

Platelet Activation ◽

Blood Cells ◽

Clinical Effect ◽

Platelet Rich Plasma ◽

Lower Number ◽

Antithrombotic Drug ◽

A Cell

SummaryRed blood cells (RBC) increase the proaggregatory capacity of a cell-free supernatant obtained by stimulating platelet-rich plasma (PRP) samples with collagen (1 μg/ml) as measured by the BASIC wave; this effect increases with the number of RBC and is proportionally greater with a lower number of platelets or when lower collagen concentrations are used.Aspirin (ASA) modifies the RBC behaviour in relation to their platelet-collagen interaction. This is demonstrated by the fact that when PRP and RBC obtained from the same subjects before and two hours after the ingestion of ASA (0.5 g) were mixed, it was found that non-AS A-RBC stimulate ASA-PRP, probably through a platelet cyclooxygenase independent pathway; ASA-RBC, however, stimulate non-ASA-PRP, but not ASA-PRP, which suggests that they may need an active platelet cyclooxygenase system for their action. This effect of ASA on RBC is not transient and was also observable 48 h after ASA ingestion. In addition, it was found that ASA-RBC greatly increase the activation of a mixture containing a small proportion of non-ASA-PRP in ASA-PRP, a situation that is expected to be encountered “in vivo” after ASA treatment. This effect of ASA-RBC on platelet activation may help to explain the sometimes contradictory clinical effect of aspirin as an antithrombotic drug.

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The Role of Polyphenols in the Modulation of Plasma Non-Enzymatic Antioxidant Capacity (NEAC)

International Journal for Vitamin and Nutrition Research ◽

10.1024/0300-9831/a000116 ◽

2012 ◽

Vol 82 (3) ◽

pp. 228-232 ◽

Cited By ~ 7

Author(s):

Mauro Serafini ◽

Giuseppa Morabito

Keyword(s):

Antioxidant Capacity ◽

Dietary Intervention ◽

Ex Vivo ◽

The Body ◽

Dietary Polyphenols ◽

Enzymatic Antioxidant ◽

Endogenous Antioxidant

Dietary polyphenols have been shown to scavenge free radicals, modulating cellular redox transcription factors in different in vitro and ex vivo models. Dietary intervention studies have shown that consumption of plant foods modulates plasma Non-Enzymatic Antioxidant Capacity (NEAC), a biomarker of the endogenous antioxidant network, in human subjects. However, the identification of the molecules responsible for this effect are yet to be obtained and evidences of an antioxidant in vivo action of polyphenols are conflicting. There is a clear discrepancy between polyphenols (PP) concentration in body fluids and the extent of increase of plasma NEAC. The low degree of absorption and the extensive metabolism of PP within the body have raised questions about their contribution to the endogenous antioxidant network. This work will discuss the role of polyphenols from galenic preparation, food extracts, and selected dietary sources as modulators of plasma NEAC in humans.

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The Role of In Vivo and Ex Vivo Diagnostic Tools in Severe Delayed Immune-Mediated Adverse Antibiotic Drug Reactions

The Journal of Allergy and Clinical Immunology In Practice ◽

10.1016/j.jaip.2020.12.052 ◽

2021 ◽

Author(s):

Ana Copaescu ◽

Effie Mouhtouris ◽

Sara Vogrin ◽

Fiona James ◽

Kyra Y.L. Chua ◽

...

Keyword(s):

Ex Vivo ◽

Diagnostic Tools ◽

Drug Reactions ◽

Immune Mediated ◽

Antibiotic Drug

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Heparan sulfate and heparin interactions with proteins

Journal of The Royal Society Interface ◽

10.1098/rsif.2015.0589 ◽

2015 ◽

Vol 12 (110) ◽

pp. 20150589 ◽

Cited By ~ 108

Author(s):

Maria C. Z. Meneghetti ◽

Ashley J. Hughes ◽

Timothy R. Rudd ◽

Helena B. Nader ◽

Andrew K. Powell ◽

...

Keyword(s):

Heparan Sulfate ◽

Ex Vivo ◽

Sequence Specificity ◽

Structure Activity ◽

Multiple Binding Sites ◽

Multiple Binding ◽

Heparin Activity

Heparan sulfate (HS) polysaccharides are ubiquitous components of the cell surface and extracellular matrix of all multicellular animals, whereas heparin is present within mast cells and can be viewed as a more sulfated, tissue-specific, HS variant. HS and heparin regulate biological processes through interactions with a large repertoire of proteins. Owing to these interactions and diverse effects observed during in vitro , ex vivo and in vivo experiments, manifold biological/pharmacological activities have been attributed to them. The properties that have been thought to bestow protein binding and biological activity upon HS and heparin vary from high levels of sequence specificity to a dependence on charge. In contrast to these opposing opinions, we will argue that the evidence supports both a level of redundancy and a degree of selectivity in the structure–activity relationship. The relationship between this apparent redundancy, the multi-dentate nature of heparin and HS polysaccharide chains, their involvement in protein networks and the multiple binding sites on proteins, each possessing different properties, will also be considered. Finally, the role of cations in modulating HS/heparin activity will be reviewed and some of the implications for structure–activity relationships and regulation will be discussed.

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