Platelet Activation and Cytokine Production during Hypothermic Cardiopulmonary Bypass – A Possible Correlation?

1998 ◽  
Vol 80 (07) ◽  
pp. 58-64 ◽  
Author(s):  
P. Ferroni ◽  
G. Speziale ◽  
G. Ruvolo ◽  
A. Giovannelli ◽  
F. M. Pulcinelli ◽  
...  
Keyword(s):  
Cardiac Surgery ◽  
Cardiopulmonary Bypass ◽  
Inflammatory Response ◽  
Platelet Activation ◽  
Cytokine Production ◽  
Inverse Correlation ◽  
Moderate Degree ◽  
Platelet Response ◽  
Activated Platelets

SummaryCardiopulmonary bypass (CPB) is associated with impaired platelet function and a systemic inflammatory response. The present study was designed to evaluate whether any correlation between platelet activation and inflammatory response during CPB exists. The results obtained from 8 patients undergoing hypothermic CPB for cardiac surgery showed the occurrence of a moderate degree of platelet activation during CPB, demonstrated by an increase of platelet CD62P expression in correlation with an increase of β-thromboglobulin levels, with a concomitant decrease of in vitro platelet response. Plasma IL-1β levels significantly increased during CPB, with a peak between 1 and 4 h after CPB. Similarly, IL-6 levels were elevated 30 min from CPB starting, peaked at 4 h, and remained elevated after 24 h. A direct correlation was found between plasma IL-1β and IL-6 levels. A significant correlation between plasma IL-1β and β-thromboglobulin levels was also found. In turn, plasma β-thromboglobulin levels correlated with CD62P expression on activated platelets. An inverse correlation was found between in vitro platelet aggregation and plasma IL-1β or IL-6 levels. From the present results it may be speculated that platelet activation during CPB may contribute, through the release of IL-1β, to activation of endothelial cells and subsequent release of other cytokines with chemotactic and pro-inflammatory properties, thus playing an important role in the inflammatory response associated with CPB.

scholarly journals Studies with a monoclonal antibody against activated platelets: evidence that a secreted 53,000-molecular weight lysosome-like granule protein is exposed on the surface of activated platelets in the circulation

Blood ◽  
1987 ◽  
Vol 70 (3) ◽  
pp. 838-845 ◽  
Author(s):  
HK Nieuwenhuis ◽  
JJ van Oosterhout ◽  
E Rozemuller ◽  
F van Iwaarden ◽  
JJ Sixma
Keyword(s):  
Cardiopulmonary Bypass ◽  
Platelet Activation ◽  
Binding Sites ◽  
Double Labeling ◽  
Flow Cytofluorometry ◽  
Activated Platelets ◽  
Normal Individuals

Abstract To define the role of activated platelets we have attempted to prepare monoclonal antibodies specific for activated platelets. The IgG2b antibody of one of the clones, designated 2.28, was studied in more detail. Native platelets from normal individuals bound 650 125I-2.28 molecules/platelet, whereas thrombin-activated platelets bound 12,600 molecules/platelet with high affinity (4.6 nmol/L). Immunoelectrophoretic analysis revealed that 2.28 reacted with a 53,000- mol wt protein. Immunocytochemistry showed that the antigen is located in a special subclass of platelet granules in unstimulated platelets and is exposed on the surface of thrombin-activated platelets. Double- labeling studies with immunogold labels disclosed simultaneous localization of 2.28 binding sites and cathepsin D in the same granules both in megakaryocytes and endothelial cells, thereby indicating that the antigen may be localized in lysosomes. By using flow cytofluorometry, in vivo platelet activation was studied in patients undergoing cardiac surgery with cardiopulmonary bypass. Increased numbers of platelets that expressed the 2.28 antigen on their surface were observed after extracorporeal perfusion. The percentage of 2.28- positive platelets in the circulation was 3.9% +/- 2.7% (SD) in controls (n = 20), 5.5% +/- 3.0% in patients (n = 10) before cardiopulmonary bypass surgery, 24.6% +/- 13.5% after the bypass, and 8.5% in two patients with acute deep venous thrombosis. These data indicate that 2.28 may serve as a useful probe of in vitro and in vivo platelet activation.

scholarly journals Studies with a monoclonal antibody against activated platelets: evidence that a secreted 53,000-molecular weight lysosome-like granule protein is exposed on the surface of activated platelets in the circulation

Blood ◽  
1987 ◽  
Vol 70 (3) ◽  
pp. 838-845 ◽  
Author(s):  
HK Nieuwenhuis ◽  
JJ van Oosterhout ◽  
E Rozemuller ◽  
F van Iwaarden ◽  
JJ Sixma
Keyword(s):  
Cardiopulmonary Bypass ◽  
Platelet Activation ◽  
Binding Sites ◽  
Double Labeling ◽  
Flow Cytofluorometry ◽  
Activated Platelets ◽  
Normal Individuals

To define the role of activated platelets we have attempted to prepare monoclonal antibodies specific for activated platelets. The IgG2b antibody of one of the clones, designated 2.28, was studied in more detail. Native platelets from normal individuals bound 650 125I-2.28 molecules/platelet, whereas thrombin-activated platelets bound 12,600 molecules/platelet with high affinity (4.6 nmol/L). Immunoelectrophoretic analysis revealed that 2.28 reacted with a 53,000- mol wt protein. Immunocytochemistry showed that the antigen is located in a special subclass of platelet granules in unstimulated platelets and is exposed on the surface of thrombin-activated platelets. Double- labeling studies with immunogold labels disclosed simultaneous localization of 2.28 binding sites and cathepsin D in the same granules both in megakaryocytes and endothelial cells, thereby indicating that the antigen may be localized in lysosomes. By using flow cytofluorometry, in vivo platelet activation was studied in patients undergoing cardiac surgery with cardiopulmonary bypass. Increased numbers of platelets that expressed the 2.28 antigen on their surface were observed after extracorporeal perfusion. The percentage of 2.28- positive platelets in the circulation was 3.9% +/- 2.7% (SD) in controls (n = 20), 5.5% +/- 3.0% in patients (n = 10) before cardiopulmonary bypass surgery, 24.6% +/- 13.5% after the bypass, and 8.5% in two patients with acute deep venous thrombosis. These data indicate that 2.28 may serve as a useful probe of in vitro and in vivo platelet activation.

scholarly journals Effect of sevoflurane on the inflammatory response during cardiopulmonary bypass in cardiac surgery: the study protocol for a randomized controlled trial

Trials ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Thiago Augusto Azevedo Maranhão Cardoso ◽  
Gudrun Kunst ◽  
Caetano Nigro Neto ◽  
José de Ribamar Costa Júnior ◽  
Carlos Gustavo Santos Silva ◽  
...  
Keyword(s):  
Randomized Controlled Trial ◽  
Cardiac Surgery ◽  
Cardiopulmonary Bypass ◽  
Inflammatory Response ◽  
Controlled Trial ◽  
Artery Bypass ◽  
Bypass Graft ◽  
Volatile Anesthetic ◽  
Serum Levels ◽  
Randomized Controlled

Abstract Background Recent experimental evidence shows that sevoflurane can reduce the inflammatory response during cardiac surgery with cardiopulmonary bypass. However, this observation so far has not been assessed in an adequately powered randomized controlled trial. Methods We plan to include one hundred patients undergoing elective coronary artery bypass graft with cardiopulmonary bypass who will be randomized to receive either volatile anesthetics during cardiopulmonary bypass or total intravenous anesthesia. The primary endpoint of the study is to assess the inflammatory response during cardiopulmonary bypass by measuring PMN-elastase serum levels. Secondary endpoints include serum levels of other pro-inflammatory markers (IL-1β, IL-6, IL-8, TNFα), anti-inflammatory cytokines (TGFβ and IL-10), and microRNA expression in peripheral blood to achieve possible epigenetic mechanisms in this process. In addition clinical endpoints such as presence of major complications in the postoperative period and length of hospital and intensive care unit stay will be assessed. Discussion The trial may determine whether adding volatile anesthetic during cardiopulmonary bypass will attenuate the inflammatory response. Trial registration ClinicalTrials.gov NCT02672345. Registered on February 2016 and updated on June 2020.

Modulation of Systemic Inflammatory Response after Cardiac Surgery

2005 ◽  
Vol 13 (4) ◽  
pp. 382-395 ◽  
Author(s):  
Shahzad G Raja ◽  
Gilles D Dreyfus
Keyword(s):  
Cardiac Surgery ◽  
Cardiopulmonary Bypass ◽  
Inflammatory Response ◽  
Inflammatory Reaction ◽  
Therapeutic Strategies ◽  
Systemic Inflammatory Response ◽  
Whole Body ◽  
Hemodynamic Stability ◽  
Immunomodulatory Agents ◽  
Considerable Research

Cardiac surgery and cardiopulmonary bypass initiate a systemic inflammatory response largely determined by blood contact with foreign surfaces and the activation of complement. It is generally accepted that cardiopulmonary bypass initiates a whole-body inflammatory reaction. The magnitude of this inflammatory reaction varies, but the persistence of any degree of inflammation may be considered potentially harmful to the cardiac patient. The development of strategies to control the inflammatory response following cardiac surgery is currently the focus of considerable research efforts. Diverse techniques including maintenance of hemodynamic stability, minimization of exposure to cardiopulmonary bypass circuitry, and pharmacologic and immunomodulatory agents have been examined in clinical studies. This article briefly reviews the current concepts of the systemic inflammatory response following cardiac surgery, and the various therapeutic strategies being used to modulate this response.

A Significant Fraction of ADAMTS13 Activity Is Associated with Activated Platelets and Their Microparticles (PMP): Implication for Regulating ADAMTS13 Activity.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1066-1066
Author(s):  
Wenche Jy ◽  
Andrew Lin ◽  
Loreta Bidot ◽  
Jaehoon Bang ◽  
Eugene Ahn ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Control Level ◽  
In Vitro Study ◽  
Ionophore A23187 ◽  
Adamts13 Activity ◽  
Activated Platelets ◽  
Membrane Bound ◽  
Low Levels

Abstract BACKGROUND: Deficiency of ADAMTS13, vWF cleaving protease, is known to be associated with TTP and some other microangiopathies, but low levels were reported in other diseases such as ITP, DIC, lupus and other thromboses. Although inhibitory autoantibodies were demonstrated in TTP, the mechanisms underlying reduced levels of ADAMTS13 in other disorders remains unclear. We tested the hypothesis that ADAMTS13 is associated with cell membranes and derived microparticles, especially from activated platelet and their microparticles (PMP), which could modulate the enzyme activities of ADAMTS13. METHODS: PRP was prepared by centrifuging citrated normal blood for 10 min at 160×g, and PPP by further centrifuging for 10 min at 3,000×g, and particle-free plasma (PFP) by further centrifuging for 15 min at 20,000×g. ADAMTS13 activity was assayed by the FRETS-VWF73 method of Kokame et al [Br J Haematol 129:93, 2005] using the Fluoroskan Ascent plate reader. Platelets were activated by ADP (10 μM) or ionophore A23187 (2 μM). RESULTS: The ADAMTS13 activity (A) of pooled PPP of 10 controls was defined as 100%. (1) In vitro study: (1a) ADAMTS13 activity was not significantly different between PPP and resting PRP. However, if the platelets in PRP were first activated by ADP for 1hr, a significant reduction of activity was observed (A = 85 ±7%, p<0.05). If the activated platelets were removed, the activity of the supernatant fell to 79 ±10% p<0.05) of the control level, and was further reduced by higher centrifugation to remove PMP (A = 66 ±12%, p<0.01). (1b) Activation by A23187, a stronger agonist producing 2–3 fold more PMP than ADP (confirmed by flow cytometry), induced a more dramatic reduction in PRP (A = 78±8%, p<0.01), and after removal of platelets (A = 71 ±11%, p<0.01), and after removal of PMP (48 ±11%, p<0.01). (1c) Interestingly, resuspending the activated platelets did not restore ADAMTS13 activity, although resuspending the PMP did partially restore the activity. (2) In vivo study: PPP from 13 patients (6 ITP, 4 APS, 3 lupus) were analyzed. The majority (11/13) of PPP samples lost activity after removal of PMP (A = 79 ±12% in PPP vs. 64 ±11% in PFP; p <0.02). CONCLUSION: These data show that a significant but variable fraction of ADAMTS13 activity is associated with activated platelets and PMP. This has several implications. First, distinguishing soluble from membrane-bound ADAMTS13 may lead to better correlation of activities with clinical findings, and may help explain low levels of ADAMTS13 in some disorders associated with platelet activation and high PMP. Second, this interaction may play a role in regulating ADAMTS13 activity. Third, membrane-bound ADAMTS13 may clear more readily from circulation, therefore inhibiting platelet activation or MP formation may have benefits for the management of microangiopathies.

The systemic inflammatory response syndrome following cardiac surgery: different expression of proinflammatory cytokines and procalcitonin in patients with and without multiorgan dysfunctions

Perfusion ◽  
2002 ◽  
Vol 17 (2) ◽  
pp. 103-109 ◽  
Author(s):  
Armin Sablotzki ◽  
Ivar Friedrich ◽  
Jörg Mühling ◽  
Marius G Dehne ◽  
Jan Spillner ◽  
...  
Keyword(s):  
Cardiac Surgery ◽  
Cardiopulmonary Bypass ◽  
Systemic Inflammatory Response Syndrome ◽  
Inflammatory Response ◽  
Heart Surgery ◽  
Open Heart Surgery ◽  
Multiple Organ ◽  
Systemic Inflammatory Response ◽  
Apache Ii Score ◽  
Organ Dysfunctions

Cardiopulmonary bypass is associated with an injury that may cause pathophysiological changes in the form of systemic inflammatory response syndrome (SIRS) or multiple organ dysfunction syndrome (MODS). In the present study, we investigated the inflammatory response of patients with multiple organ dysfunctions following open-heart surgery. Plasma levels of cytokines (IL-1β, IL-6, IL-8, IL-18) and procalcitonin (PCT) were measured on the first four postoperative days in 12 adult male patients with SIRS and two or more organ dysfunctions after myocar-dial revascularization (MODS group), and 15 patients without organ dysfunctions (SIRS group). All cytokines (except IL-1β) and PCT were significantly elevated in MODS patients, with peak values at the first two postoperative days. The results of our study show a different expression of members of the IL-1 family following extracorporeal circulation. For the first time, we can document that IL-18 is involved in the inflammatory response and the initiation of the MODS following cardiopulmonary bypass. In addition to APACHE-II score, PCT, IL-8, and IL-18 may be used as parameters for the prognosis of patients with organ dysfunctions after cardiac surgery. Furthermore, it must be noted that the duration of the surgical procedure is one of the most important factors for the initiation of the inflammatory response.

scholarly journals Influence of lithium chloride on neutrophil activation in the development of systemic inflammatory response syndrome in patients after on-pump cardiac surgery

2020 ◽  
pp. 47-53
Author(s):  
О.А. Гребенчиков ◽  
И.С. Касаткина ◽  
А.Н. Кузовлев ◽  
А.В. Лобанов ◽  
А.В. Ершов
Keyword(s):  
Cardiac Surgery ◽  
Systemic Inflammatory Response Syndrome ◽  
Inflammatory Response ◽  
Blood Serum ◽  
Lithium Chloride ◽  
Human Neutrophils ◽  
Spontaneous Apoptosis ◽  
Chloride Solutions ◽  
Activated Neutrophils

Цель исследования - изучение in vitro действия хлорида лития на активность нейтрофилов человека при действии сывороток пациентов с синдромом системного воспалительного ответа, развившемся после операций на сердце с искусственным кровообращением. Методика. Исследование проводили in vitro на нейтрофилах, выделенных из крови 6 здоровых доноров. Нейтрофилы активировали при помощи сыворотки пациентов с синдромом системного воспалительного ответа (ССВО), перенесших операции на сердце с искусственным кровообращением (ИК). Активность нейтрофилов оценивали с использованием флуоресцентных антител к маркерам дегрануляции CD11b и CD66b. Уровень апоптоза и некроза нейтрофилов оценивали через 22 ч после выделения из крови здоровых доноров; количественная оценка была проведена с использованием аннексина V и иодистого пропидия на проточном цитофлуориметре. Интактные и активированные нейтрофилы обрабатывали раствором хлорида лития в концентрациях 0,3; 3,0 и 9,0 мМ. Результаты. Инкубация нейтрофилов с сывороткой крови пациентов с ССВО после операций на сердце с ИК увеличивала экспрессию CD11b в 1,5 раза и экспрессию CD66b в 1,4 раза в сравнении с экспрессией на интактных нейтрофилах. Инкубация нейтрофилов с сывороткой крови пациентов с ССВО и раствором хлорида лития в концентрациях 3,0 и 9,0 мМ приводило к статистически значимому снижению уровня экспрессии CD11b CD66b на поверхности нейтрофилов в сравнении с активированными контрольными. Установлено, что хлорид лития в концентрациях 3,0 и 9,0 мМ возвращал уровни экспрессии CD11b и CD66b на активированных нейтрофилах к уровню экспрессии на интактных нейтрофилах. В концентрации 0,3 мМ хлорид лития, используемый при инкубации с активированными нейтрофилами, не вызывал значимого снижения экспрессии CD11b и CD66b относительно контрольных активированных нейтрофилов. Экспрессия CD11b и CD66b на активированных нейтрофилах при их инкубации с хлоридом лития в концентрации 0,3 мМ была значимо выше относительно экспрессии данных молекул на интактных нейтрофилах. Сыворотка пациентов с развившемся ССВО снижала спонтанный апоптоз нейтрофилов, а раствор хлорида лития в концентрации 3,0 или 9,0 мМ, добавленный в среду инкубации, увеличивал способность нейтрофилов к спонтанному апоптозу. Заключение. Хлорид лития оказывал противовоспалительный эффект снижал дегрануляцию и активацию нйтрофилов посредством уменьшения уровня экспрессии молекул CD11b и CD66b на поверхности нейтрофилов, которые предварительно были активированы сыворотками пациентов с ССВО. В концентрации 3,0 мМ и выше хлорид лития индуцировал спонтанный апоптоз нейтрофилов, активированных сыворотками пациентов с ССВО после операций на сердце с ИК. The aim of this work was to study the anti-inflammatory effect of lithium chloride on human neutrophils in vitro under the action of the serum of patients with systemic inflammatory response syndrome (SIRS), which developed after on-pump cardiac surgery. Methods. The study was performed on neutrophils isolated from the blood of five healthy donors, which was activated using serum from patients with SIRS. Neutrophil activity was assessed using fluorescent antibodies to CD11b and CD66b degranulation markers. The level of apoptosis and necrosis of human neutrophils was evaluated 22 hours after isolation. Quantification was performed using annexin V and propidium iodide on a flow cytometer. Intact and activated neutrophils were treated with 0.3, 3.0 аnd 9.0 mM lithium chlorides. Results. Incubation of neutrophils with the blood serum of patients with SIRS after on-pump cardiac surgery increased the expression of CD11b by 1.5 times and the expression of CD66b by 1.4 times compared to expression on intact neutrophils. Incubation of neutrophils with blood serum of patients with SIRS and 3.0 and 9.0 mM lithium chloride solutions led to a statistically significant decrease in the level of expression of CD11b CD66b on the surface of neutrophils in comparison with control activated neutrophils. It was found that 3.0 and 9.0 mM lithium chloride solutions returned the expression levels of CD11b and CD66b on activated neutrophils to the expression level on intact neutrophils. 0.3 mM of lithium chloride, used during incubation with activated neutrophils, did not cause a significant decrease in the expression of CD11b and CD66b relative to control activated neutrophils. The expression of CD11b and CD66b on activated neutrophils during their incubation with 0.3 mM of lithium chloride was significantly higher relative to the expression of these molecules on intact neutrophils. The serum of patients with advanced SIRS decreased the ability of neutrophils to spontaneous apoptosis. 3.0 or 9.0 mM lithium chloride solutions added to the incubation medium increased the ability of neutrophils to spontaneous apoptosis. Conclusion. Lithium chloride reduced the degranulation and activation of neutrophils by reducing the expression level of CD11b and CD66b molecules on the surface of neutrophils that were previously activated by the serum of patients with SIRS. This effect determines the anti-inflammatory influence of lithium chloride. Lithium chloride at 3.0 mM and higher induced spontaneous apoptosis of neutrophils activated by the serum of patients with SIRS after on-pump cardiac surgery.

scholarly journals Direct detection of activated platelets and platelet-derived microparticles in humans

Blood ◽  
1990 ◽  
Vol 75 (1) ◽  
pp. 128-138 ◽  
Author(s):  
CS Abrams ◽  
N Ellison ◽  
AZ Budzynski ◽  
SJ Shattil
Keyword(s):  
Flow Cytometry ◽  
Cardiopulmonary Bypass ◽  
Platelet Activation ◽  
Bleeding Time ◽  
Relative Proportion ◽  
Open Heart Surgery ◽  
Normal Range ◽  
In Vivo Models ◽  
Platelet Surface ◽  
Activated Platelets

Flow cytometry was used to determine whether activated platelets and platelet-derived microparticles can be detected directly in whole blood after a hemostatic insult. Two different in vivo models of platelet activation were examined: (1) a standardized bleeding time, and (2) cardiopulmonary bypass. Platelets and microplatelets were identified with a biotinylated anti-glycoprotein (GP)lb antibody and a fluorophore, phycoerythrin-streptavidin. Microparticles were distinguished from platelets by light scatter. Activated platelets were detected with three fluorescein-labeled monoclonal antibodies (MoAbs): (1) PAC1, which binds to the activated form of GPIIb-IIIa; (2) 9F9, a newly developed antibody that is specific for fibrinogen bound to the surface of activated platelets; and (3) S12, which binds to an alpha- granule membrane protein expressed on the platelet surface after granule secretion. In nine normal subjects, bleeding times ranged from 4.5 to 7.5 minutes. Over this time, there was a progressive increase in the amount of PAC1, 9F9, and S12 bound to platelets in blood emerging from the bleeding time wound. With all three antibodies, platelet activation was apparent as early as 30 seconds after the incision (P less than .03). Activation was accompanied by a progressive decrease in the concentration of platelets in blood from the wound, while the concentration of microparticles increased slightly. In nine patients undergoing open heart surgery, 1 hour of cardiopulmonary bypass caused a 2.2-fold increase in the relative proportion of microparticles in circulating blood (P less than .001). Moreover, bypass caused platelet activation as evidenced by a mean two- to threefold increase in PAC1 binding to platelets. Although this increase was significant (P less than .02), PAC1 binding exceeded the normal range for unstimulated control platelets in only 5 of 9 patients, and 9F9 and S12 binding exceeded the normal range in only two patients. Taken together, these studies demonstrate that it is now feasible using flow cytometry to evaluate the extent of platelet activation and the presence of platelet- derived microparticles in the circulation of humans.

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