CD11b expression on polymorphonuclear leukocytes from patients with ankylosing spondylitis in a lipopolysaccharide-stimulated whole blood ex vivo model

2013 ◽  
Vol 20 (1) ◽  
pp. 60-67 ◽  
Author(s):  
Gabriela P. Mathias ◽  
Magno D. Garcia ◽  
Fabricio S. Neves
Keyword(s):  
Ankylosing Spondylitis ◽  
Whole Blood ◽  
Polymorphonuclear Leukocytes ◽  
Ex Vivo ◽  
Ex Vivo Model ◽  
Cd11b Expression

Determinants of platelet conjugate formation with polymorphonuclear leukocytes or monocytes in whole blood

2007 ◽  
Vol 98 (12) ◽  
pp. 1276-1284 ◽  
Author(s):  
Simona Costanzo ◽  
Branislav Vohnout ◽  
Licia Iacoviello ◽  
Giovanni de Gaetano ◽  
Benedetta Izzi ◽  
...  
Keyword(s):  
Platelet Count ◽  
Whole Blood ◽  
Polymorphonuclear Leukocytes ◽  
Large Population ◽  
Mixed Cell ◽  
Cd11b Expression ◽  
Glucose Levels ◽  
Primary Platelet ◽  
Collagen Stimulation

SummaryFollowing preliminary in-vitro experiments, platelet-leukocyte conjugates and their determinants were evaluated in citrated whole blood from 349 subjects (209 women, age 16–92 years) randomly recruited from the general population. Platelet activation by ADP/collagen but not leukocyte stimulation by fMLP or LTB4 resulted in formation of platelet conjugates with PMN or monocytes. In the population study, mixed cell conjugates, platelet P-selectin and leukocyte CD11b were measured by flow cytometry both at baseline and after in-vitro stimulation with ADP/collagen. The latter significantly increased platelet conjugates with either PMN or monocytes, platelet P-selectin and leukocyte CD11b expression. Platelet count significantly correlated with platelet-PMN, platelet-monocyte conjugates and P-selectin both at baseline and upon stimulation. In all conditions, both conjugate levels correlated with each other, when adjusted for gender, age and platelet count. Age correlated with platelet-PMN conjugate numbers in basal and stimulated conditions and with basal P-selectin. ADP/collagen stimulation resulted in higher P-selectin and conjugates values in women. Among risk factors, a significant correlation was found between conjugate and glucose levels. In conclusion, the presence and formation in whole blood from a large population of plateletleukocyte conjugates reflects primary platelet – but not leukocyte – activation and varies with gender, age, platelet count and blood glucose.

Regulation of tissue factor-induced coagulation and platelet aggregation in flowing whole blood

2005 ◽  
Vol 93 (01) ◽  
pp. 97-105 ◽  
Author(s):  
Marijke Kuijpers ◽  
Cécile Nieuwenhuys ◽  
Marion Feijge ◽  
Willem Kloots ◽  
Peter Giesen ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Whole Blood ◽  
Platelet Adhesion ◽  
Ex Vivo ◽  
Aggregate Formation ◽  
Ex Vivo Model ◽  
Platelet Surface ◽  
Inhibiting Effect ◽  
Activated Platelets ◽  
Adp Receptor

SummaryPhotochemically induced thrombosis (a thrombin-dependent process) was measured in rats treated with moderate doses of anticoagulants, but which appeared to be unchanged. We considered the possibility that platelet-inhibiting agents, which also indirectly inhibit coagulation, would act as more potent antithrombotic agents.Inhibitors used as such were prostaglandin E1 (PGE1), which elevates cyclic AMP levels, and the P2Y12 ADP-receptor antagonist,AR-C69931MX. Effects of these agents were investigated in an ex vivo model system, in which whole blood under coagulant conditions was perfused over fibrinogen at defined wall shear rate. Perfusion of blood (rat or human) in the presence of tissue factor resulted in deposition of activated platelets and subsequent aggregate formation, along with exposure of procoagulant phosphatidylserine (PS) on the platelet surface and formation of fibrin fibers. In the presence of PGE1 aggregation was completely inhibited,but platelet adhesion and PS exposure were only party reduced, while fibrin formation was hardly affected. Treatment with AR-C69931MX caused similar, but less complete effects.These results indicate that in tissue factor- triggered blood under conditions of flow:(i) the platelet procoagulant response is independent of aggregate formation; (ii) the platelet-inhibiting effect of PGE1 and AR-C69931MX is sufficient to suppress aggregation, but not platelet adhesion and coagulation. These platelet inhibitors thus maintain their aggregation- inhibiting effect at sites of thrombin formation.

Modulation of Blood Cell Activation by Four Commonly Used Anticoagulants

1997 ◽  
Vol 77 (04) ◽  
pp. 690-696 ◽  
Author(s):  
Charlotte Sissener Engstad ◽  
Tore Jarl Gutteberg ◽  
Bjarne Østerud
Keyword(s):  
Whole Blood ◽  
Cell Activation ◽  
Ex Vivo ◽  
Ex Vivo Model ◽  
Platelet Factor ◽  
Human Whole Blood ◽  
Platelet Parameter ◽  
Factor Α ◽  
Edta Blood

SummaryIn the past years, our group has made several observations suggesting that blood cells behave differently and when stimulated, release different levels of cytokines, depending on which anticoagulant the blood has been drawn into. The aim of this study was therefore to compare the effect of the four anticoagulants EDTA, citrate, heparin and hirudin on monocyte, neutrophil (PMN), and platelet function in human whole blood. Human whole blood was employed as an ex vivo model of cytokine production and protein secretion, and lipopolysaccharide (LPS) induced tissue factor (TF) activity in monocytes and LPS induced tumor necrosis factor α (TNFα) release were chosen as parameters of monocyte activation. Platelet factor 4 (PF4) secretion and LPS induced lacto ferrin release were chosen as parameters of platelet and PMN activation, respectively. When human whole blood was stimulated with 5 ng/ml LPS for 2 h, TF activity in monocytes isolated from EDTA blood was found to be 2.9 mU/106 cells, whereas TF activity in monocytes isolated from citrated, heparinized and hirudinized blood was 14.7, 24.7 and 28.5 mU/106 cells, respectively. TNFα concentrations in platelet poor plasma (PPP) isolated from whole blood stimulated with 5 ng/ml LPS for 2 h was increased with 200,400 and 350% in citrated, heparinized and hirudinized blood respectively, as compared to EDTA blood. Next, the effect of the anticoagulant on PMN secretion was measured. PPP isolated from whole blood incubated with 5 ng/ml LPS for 90 min contained 1170 ng/ml (EDTA blood), 2880 ng/ml (citrated blood), 4220 ng/ml (heparinized blood), and 5520 ng/ml lactoferrin (hirudinized blood). When studying the platelet parameter PF4, whole blood was incubated without any stimuli for 60 min, and we found that heparin PPP contained 1180 ng/ml PF4, while hirudin PPP contained 469 ng/ml, citrate PPP 440 ng/ml, and EDTA PPP 217 ng/ml PF4, respectively. Finally, the low molecular weight heparin compound Fragmin had no enhancing effect on PF4 levels in whole blood. It is concluded that the anticoagulant used in in vitro experiments has a large influence on the parameters measured.

Lyophilised reconstituted human platelets increase thrombus formation in a clinical ex vivo model of deep arterial injury

2012 ◽  
Vol 108 (07) ◽  
pp. 176-182 ◽  
Author(s):  
Jennifer Raftis ◽  
Andrew Lucking ◽  
Amanda Hunter ◽  
Mike Millar ◽  
Mike Fitzpatrick ◽  
...  
Keyword(s):  
Whole Blood ◽  
Ex Vivo ◽  
Thrombus Formation ◽  
Morphological Characteristics ◽  
Principal Component ◽  
Arterial Injury ◽  
Human Platelets ◽  
Dependent Manner ◽  
Ex Vivo Model ◽  
Electron Microscopic

SummaryPlatelets are the principal component of the innate haemostatic system that protect from traumatic bleeding. We investigated whether lyophilised human platelets (LHPs) could enhance clot formation within platelet-free and whole blood environments using an ex vivo model of deep arterial injury. Lyophilised human platelets were produced from stored human platelets and characterised using conventional, fluorescent and electron microscopic techniques. LHPs were resuspended in platelet-free plasma (PFP) obtained from citrated whole human blood to form final concentrations of 0,20 and 200 × 109 LHPs/L. LHPs with recalcified PFP or whole blood were perfused through the chamber at low (212 s-1) and high (1,690 s-1) shear rates with porcine aortic tunica media as thrombogenic substrate. LHPs shared morphological characteristics with native human platelets and were incorporated into clot generated from PFP or whole blood. Histomorphometrically measured mean thrombus area increased in a dose-dependent manner following the addition of LHPs to PFP under conditions of high shear [704 μm2 ± 186 μm2 (mean ± SEM), 1,511 μm2 ± 320 μm2 and 2,378 μm2 ± 315 μm2, for LHPs at 0, 20 and 200 × 109 /l, respectively (p= 0.012)]. Lyophil-ised human platelets retain haemostatic properties when reconstituted in both PFP and whole blood, and enhance thrombus formation in a model of deep arterial injury. These data suggest that LHPs have the potential to serve as a therapeutic intervention during haemorrhage under circumstances of trauma, and platelet depletion or dysfunction.

A Novel Porcine Ex Vivo Retina Culture Model for Oxidative Stress Induced by H2O2

2017 ◽  
Vol 45 (1) ◽  
pp. 11-25 ◽  
Author(s):  
José Hurst ◽  
Sandra Kuehn ◽  
Adelina Jashari ◽  
Teresa Tsai ◽  
Karl Ulrich Bartz-Schmidt ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Molecular Mechanisms ◽  
Ganglion Cells ◽  
Ex Vivo ◽  
Laboratory Animals ◽  
Ex Vivo Model ◽  
Cd11b Expression ◽  
Hsp70 Mrna ◽  
New Therapeutic Approaches ◽  
Culture Models

Oxidative stress is a key player in many ophthalmic diseases. However, the role of oxidative stress in most degenerative processes is not yet known. Therefore, accurate and practical models are required to efficiently screen for therapeutics. Porcine eyes are closely related to the human eye, and can be obtained from the abattoir as a by-product of the food industry. Therefore, they offer excellent opportunities for the development of culture models with which to pre-screen potential therapies, while reducing the use of laboratory animals. To induce oxidative stress, organotypic cultures of porcine retina were treated with different doses of hydrogen peroxide (H2O2; 100, 300 and 500μM) for three hours. On days 3 and 8, the retinas were conserved for histological and Western blotting analyses and for evaluation of gene expression, which determined the number of retinal ganglion cells (RGCs), the activation state of glial cells, and the expression levels of several oxidative stress markers. H2O2 treatment led to a reduction in the number of RGCs and to an increase in apoptotic RGCs. In addition, a dose-dependent increase of microglia and an elevation of CD11b expression was observed. On day 3, a reduction of IL-1β, and an increase of iNOS, as well as of HSP70 mRNA were found. On day 8, an increase in TNF-α and IL-1β mRNA expression was detected. In conclusion, this ex vivo model offers an opportunity to study the molecular mechanisms underlying certain eye disorders and to test new therapeutic approaches to diminish the effects of oxidative stress.

Fourier Transform Infrared Spectroscopy of Protein Adsorption from Whole Blood: Ex Vivo Dog Studies

1981 ◽  
Vol 35 (4) ◽  
pp. 353-357 ◽  
Author(s):  
R. M. Gendreau ◽  
S. Winters ◽  
R. I. Leininger ◽  
D. Fink ◽  
C. R. Hassler ◽  
...  
Keyword(s):  
Fourier Transform ◽  
Protein Adsorption ◽  
Whole Blood ◽  
Experimental Approach ◽  
Ex Vivo ◽  
Fourier Transform Infrared ◽  
Ex Vivo Model ◽  
Flowing Blood ◽  
The Relationship ◽  
Exact Relationship

A Fourier transform infrared/attenuated total reflectance technique has been developed to study protein adsorption onto surfaces. The application of this technique to an ex vivo model using a beagle dog as the source of whole, flowing blood is described (currently, high-quality infrared spectra are being collected at 5-s intervals of protein adsorption). This approach has enabled the authors to identify albumin and glycoproteins as the initially adsorbing species, with the subsequent competitive replacement of part of this protein layer with fibrinogen and other proteins. The exact relationship between the pattern of protein adsorption from whole blood and the generation of a thrombus (clot) is not yet clear, but it is hoped that this type of experimental approach will help clarify the relationship.

Clopidogrel inhibits platelet-leukocyte adhesion and plateletdependent leukocyte activation

2005 ◽  
Vol 94 (09) ◽  
pp. 568-577 ◽  
Author(s):  
Stefano Manarini ◽  
Giuseppe Dell’Elba ◽  
Nicola Martelli ◽  
Emanuela Napoleone ◽  
Angelomaria Di Santo ◽  
...  
Keyword(s):  
Whole Blood ◽  
Active Metabolite ◽  
Polymorphonuclear Leukocytes ◽  
Ex Vivo ◽  
Leukocyte Adhesion ◽  
Human Platelets ◽  
Cell System ◽  
Anti Platelet Therapy ◽  
Acute Complications

SummaryClopidogrel is considered to be an important therapeutic advance in anti-platelet therapy. We investigated whether inhibition by clopidogrel results in a reduced capacity of platelets to adhere and stimulate pro-atherothrombotic and inflammatory functions in polymorphonuclear leukocytes (PMN) and in monocytes (MN). An eventual effect on these processes could further substantiate anti-atherothrombotic properties of this drug. The effects of clopidogrel or of its active metabolite were investigated on ADP or thrombin receptor-induced platelet activation and on platelet-leukocyte interactions ex vivo in the mouse or in vitro in isolated human cells or whole blood, respectively. Clopidogrel inhibited platelet aggregation, expression of P-selectin, platelet-PMN adhesion and platelet-dependent ROS production in mouse PMN. Similarly pretreatment of human platelets with the active metabolite of clopidogrel in vitro resulted in a profound inhibition of platelet P-selectin expression, platelet-PMN adhesion and production of ROS by PMN. Pretreatment with the active metabolite of clopidogrel significantly impaired the ability of platelets to up-regulate the expression of TF procoagulant activity in MN, in a washed cell system. Moreover, the active metabolite of clopidogrel inhibited rapidTF exposure on platelet as well as on leukocyte surfaces in whole blood. By reducing platelet-dependent up-regulation of inflammatory and pro-atherothrombotic functions in leukocytes, clopidogrel may reduce inflammation that underlies the chronic process of atherosclerosis and its acute complications.

scholarly journals Effect of Cannabidiol (CBD) on Canine Inflammatory Response: An Ex Vivo Study on LPS Stimulated Whole Blood

2021 ◽  
Vol 8 (9) ◽  
pp. 185
Author(s):  
Enrico Gugliandolo ◽  
Patrizia Licata ◽  
Alessio Filippo Peritore ◽  
Rosalba Siracusa ◽  
Ramona D’Amico ◽  
...  
Keyword(s):  
Veterinary Medicine ◽  
Inflammatory Response ◽  
Whole Blood ◽  
Ex Vivo ◽  
Biological Effects ◽  
Treated Group ◽  
Ex Vivo Model ◽  
Group A ◽  
Anti Inflammatory ◽  
First Time

The use of cannabidiol (CBD) for animal species is an area of growing interest, for example for its anti-inflammatory and immuno-modulating properties, even though all of its biological effects are still not fully understood, especially in veterinary medicine. Therefore, the aim of this study was to investigate the anti-inflammatory and immuno-modulating properties of CBD for the first time directly in canine inflammatory response. We used an ex vivo model of LPS-stimulated whole dog blood. We stimulated the whole blood from healthy dogs with LPS 100 ng/mL for 24 h in the presence or not of CBD 50 and 100 μg/mL. We observed a reduction in IL-6 and TNF-α production from the group treated with CBD, but non-altered IL-10 levels. Moreover, we also observed from the CBD-treated group a reduction in Nf-κB and COX-2 expression. In conclusion, we demonstrated for the first time the anti-inflammatory and immuno-modulating properties of CBD directly in dogs’ immune cells, using a canine ex vivo inflammatory model. The results obtained from these studies encourage further studies to better understand the possible therapeutic role of CBD in veterinary medicine.

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