Low degree of activation of circulating neutrophils determined by flow cytometry during cardiac surgery with cardiopulmonary bypass

SummaryCardiac surgery is complicated by the occurrence of post-operative bleeding due to platelet dysfunction. This is largely caused by platelet activation and consumption during cardiopulmonary bypass. Patients undergoing cardiac surgery requiring cardiopulmonary bypass were studied to determine whether early platelet changes due to bypass could be inhibited using the platelet-selective nitric oxide donor S-nitroso-glutathione (GSNO). Flow cytometry was used to measure platelet surface expression of P-selectin (an α-granule protein) and glycoproteins (GP) IIb/IIIa and Ib (mediators of aggregation and adhesion) before and 5 and 10 min after commencing cardiopulmonary bypass, in 6 controls and 6 patients receiving GSNO 50 μg/min. Platelet P-selectin expression increased during bypass both in controls and patients receiving GSNO. Glycoproteins IIb/IIIa and Ib fell during bypass in control and GSNO-treated patients. There was no difference between control and GSNO-treated groups. Thus no significant platelet inhibition by S-nitrosoglutathione was demonstrated under these conditions.

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Expression of CD200/CD200R regulatory molecules on granulocytes and monocytes is modulated by cardiac surgical operation

Perfusion ◽

10.1177/0267659110381451 ◽

2010 ◽

Vol 25 (6) ◽

pp. 389-397 ◽

Cited By ~ 9

Author(s):

Jan Krejsek ◽

Martina Kolackova ◽

Jiri Mandak ◽

Pavel Kunes ◽

Karolina Jankovicova ◽

...

Keyword(s):

Flow Cytometry ◽

Cardiac Surgery ◽

Cardiopulmonary Bypass ◽

Inflammatory Response ◽

Early Postoperative Period ◽

Regulatory Mechanisms ◽

Surgical Patients ◽

Surgical Operation ◽

Significant Difference ◽

Time Point

Aims: Cardiac surgical operation is inseparably linked to the induction of an inflammatory response. Both humoral and cellular regulatory mechanisms are operating to maintain body homeostasis. We followed the changes in the expression of CD200/CD200R regulatory molecules on monocytes and granulocyte of cardiac surgical patients operated on using either standard (OP) or modified “mini-invasive” cardiopulmonary bypass (MOP). Methods: Expression of CD200/CD200R regulatory molecules was determined by flow cytometry. Results: The expression of CD200R on granulocytes was increased after surgery in both groups of patients, but the increase was statistically significant only in OP patients (p<0.01). At this time point, there was a significant difference in CD200R expression on granulocytes when comparing OP to MOP patients, being higher in the former group (p<0.01). The expression of CD200R on monocytes was diminished after surgery and during an early postoperative period in both groups of patients. The expression of CD200 on monocytes was significantly diminished after surgery in both groups (p<0.01). Nonetheless, we observed an increase in CD200 expression in OP patients at the 3rd postoperative day. There was a statistically significantly increased CD200 expression on monocytes of OP patients (p<0.001) at the 3rd postoperative day when we compared OP and MOP groups. The expression of CD200 on granulocytes was significantly higher after surgery and at the 3rd postoperative day in OP when compared to MOP patients. Conclusions: CD200R expression on granulocytes was significantly increased, while CD200 and CD200R expression on monocytes was decreased after cardiac surgery.

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Up-regulation of the Apo/Fas (CD95) complex on neutrophils harvested during cardiac surgery: distinct findings in patients operated on with or without the use of cardiopulmonary bypass

Perfusion ◽

10.1177/0267659110363318 ◽

2010 ◽

Vol 25 (1) ◽

pp. 41-46 ◽

Cited By ~ 6

Author(s):

Pavel Kunes ◽

Jiri Mandak ◽

Jan Harrer ◽

Martina Kolackova ◽

Ctirad Andrys ◽

...

Keyword(s):

Flow Cytometry ◽

Cardiac Surgery ◽

Cardiopulmonary Bypass ◽

Similar Pattern ◽

Baseline Level ◽

Direct Evidence ◽

Percentage Change ◽

Surgical Patients ◽

Off Pump

Aims: In a group of patients undergoing cardiac surgery performed both with (“on-pump”) and without the use (“offpump”) of cardiopulmonary bypass (CPB), we studied the changes of neutrophil membrane apoptosis-inducing complex Apo/Fas. Methods: Expression of Apo/Fas (CD95) on leukocytes was evaluated by flow cytometry. Results: In “on-pump” patients, we found an increase in the expression of CD95 median intensity fluorescence (MFI) on granulocytes from a baseline level median=56, (Q 1=45.5, Q3=64) to a median=88, (Q1=62, Q 3=109.5; p<0.01) at the 3rd postoperative day and median=74, (Q1=63, Q3=84.5; p<0.01) at the 7th postoperative day. In “off-pump” patients, granulocyte CD95 MFI was median=55, (Q1=51, Q3=84) before surgery. The significant increase was found on the 3rd postoperative day only; median=90, (Q 1=66; Q3=98; p<0.05). A similar pattern in the CD95 expression was also found if percentage changes of granulocyte CD95 MFI were followed. Moreover, the significantly increased Apo/Fas expression expressed as a percentage change of CD95 MFI was found in “on-pump” patients compared to “off-pump” patients, both at the 3rd postoperative day (p<0.05) and at the 7th postoperative day (p<0.01). Conclusions: This is the first direct evidence of increasing densities of the Apo/Fas complex on neutrophils in cardiac surgical patients.

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Detection of leukocyte filtration and potential selective migration during use of cardiopulmonary bypass in cardiac surgery by flow cytometry

10.1117/12.349200 ◽

1999 ◽

Cited By ~ 1

Author(s):

Joerg Hambsch ◽

Veronika Schlykow ◽

Jozsef Bocsi ◽

Peter Schneider ◽

Michal Pipek ◽

...

Keyword(s):

Flow Cytometry ◽

Cardiac Surgery ◽

Cardiopulmonary Bypass ◽

Selective Migration ◽

Leukocyte Filtration

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Identification of tissue factor and platelet-derived particles on leukocytes during cardiopulmonary bypass by flow cytometry and immunoelectron microscopy

Thrombosis and Haemostasis ◽

10.1160/th07-01-0025 ◽

2007 ◽

Vol 98 (08) ◽

pp. 368-374 ◽

Cited By ~ 9

Author(s):

Jihwa Chung ◽

Hidenori Suzuki ◽

Noriyuki Tabuchi ◽

Kengo Sato ◽

Aya Shibamiya ◽

...

Keyword(s):

Flow Cytometry ◽

Cardiac Surgery ◽

Cardiopulmonary Bypass ◽

Tissue Factor ◽

Immunoelectron Microscopy ◽

Microscopic Analysis ◽

Coagulation System ◽

Flow Cytometric ◽

Platelet Antigen

SummaryThe extrinsic coagulation system initiated by tissue factor (TF) appears to be a major procoagulant stimulus during cardiopulmonary bypass (CPB), although the precise mechanisms remain to be revealed.We recently reported the appearance ofTF-bearing leukocytes during CPB and described their role in promoting coagulation. In this study, we visually identified the in-vivo appearance ofTF-bearing leukocytes and platelet-derived particles on leukocytes in the pericardial blood during cardiac surgery with CPB, by flow cytometry and immunoelectron microscopy. Preliminary flow cytometric experiments showed that the proportion of TF-positive or both TF- and platelet antigen CD41a-positive leukocytes was increased markedly in pericardial blood obtained during CPB,compared with the proportions in preoperative circulating blood. Immunoelectron microscopic analysis revealed that both monocytes and polymorphonuclear leukocytes in the pericardial blood express TF. On the surfaces of these cells, CD41a-positive or both CD41a- and TF-positive platelet-derived particles were observed. Platelet-derived particles include not only microparticles, but also platelets themselves. Leukocytes from preoperative circulating blood contained far fewer of these particles. Our results demonstrate the in-vivo appearance of TF-bearing platelet-derived particles on leukocytes during cardiac surgery with CPB.These findings may be important for the development of strategies to control procoagulant activities during and after cardiac surgery.

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Effects of conventional CPB and mini-CPB on neutrophils CD162, CD166 and CD195 expression

Perfusion ◽

10.1177/0267659116669586 ◽

2016 ◽

Vol 32 (2) ◽

pp. 141-150 ◽

Cited By ~ 5

Author(s):

Drahomira Holmannova ◽

Martina Kolackova ◽

Jiri Mandak ◽

Pavel Kunes ◽

Zdenka Holubcova ◽

...

Keyword(s):

Flow Cytometry ◽

Cardiac Surgery ◽

Cardiopulmonary Bypass ◽

Fluorescence Intensity ◽

Study Groups ◽

Median Fluorescence Intensity ◽

The Other ◽

Blood Samples ◽

Post Surgery ◽

The Impact

Objective: Cardiac surgery is known to trigger a systemic inflammatory response. While the use of conventional cardiopulmonary bypass (CPB) results in profound inflammation, modified mini-CPB is considered less harmful. We evaluated the impact of cardiac surgery on the expression of CD162, CD166, CD195 molecules and their association with the type of CPB used. Methods and Results: Twenty-four patients were enrolled in our study. Twelve of them were operated using conventional CPB while the other twelve patients underwent surgery with mini-CPB. Blood samples were analysed by flow cytometry. We observed a significant increase in median fluorescence intensity of CD162 and CD195 that peaked instantly after surgery and normalized to the baseline value on the 1st day post surgery, whereas CD166 was initially down-regulated and its median fluorescence intensity (MFI) value increased to the baseline in the next few days. Conclusion: We observed immediate changes in the expression of CD162, CD166, and CD195 molecules on the neutrophils after surgery in both study groups of patients. The intensity of the observed changes was significantly greater in the group of patients who underwent conventional CPB compared to patients who underwent mini-CPB cardiac surgery.

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Differential modulatory effects of Propofol and Sevoflurane anesthesia on blood monocyte HLA-DR and CD163 expression during and after cardiac surgery with cardiopulmonary bypass: a preliminary randomized flow cytometry study

Perfusion ◽

10.1177/0267659119848295 ◽

2019 ◽

Vol 35 (1) ◽

pp. 48-56

Author(s):

Silverio Sbrana ◽

Anna Nunziata ◽

Simona Storti ◽

Dorela Haxhiademi ◽

Annamaria Mazzone ◽

...

Keyword(s):

Flow Cytometry ◽

Cardiac Surgery ◽

Cardiopulmonary Bypass ◽

Lactate Dehydrogenase ◽

Blood Monocyte ◽

Compensatory Mechanism ◽

Monocyte Subset ◽

C Reactive Protein ◽

Reactive Protein ◽

Hla Dr

Introduction: The increase of the anti-inflammatory CD163highHLA-DRlow blood monocyte subset is one of the mechanisms dampening inflammation during cardiac surgery with cardiopulmonary bypass. We evaluated the effect of two different anesthetic protocols, intravenous Propofol infusion or Sevoflurane-gas administration, on the perioperative frequency of this subset. Methods: Blood from patients (Propofol = 11, Sevoflurane = 13) undergoing minimally invasive mitral valve surgery was drawn preoperatively (T1), before declamping (T2), at 6 (T3), 24 (T4), 48 (T5), and 72 hours (T6) after declamping. C-reactive protein, haptoglobin, and lactate dehydrogenase were measured. A hemolytic index, as C-reactive protein/haptoglobin ratio, was introduced. Monocyte expression of HLA-DR, CD163, and the CD163highHLA-DRlow subset fraction was quantified by flow cytometry. Baseline-referred variations of plasmatic and cellular data at T2 were normalized for clamping times. Subsequent time-point variations were normalized for the final cardiopulmonary bypass times. Results: Variations of hemolytic index and lactate dehydrogenase were higher with Propofol at T3 (p = 0.004 and p = 0.02, respectively) when compared with Sevoflurane. At T2, the down-modulation of CD163 was higher with Propofol (p = 0.005). Starting from T3, the up-regulatory trend of CD163 was basically higher with Propofol, although not significantly. Propofol induced higher increments of HLA-DR low fractions, at T2 (p = 0.04) and, to a lesser extent, at T4 (p = 0.06). Starting from T3, the CD163highHLA-DRlow subset variations were higher with Propofol, especially at T4 and T6. Conclusion: Propofol seems to induce a higher postoperative fraction of the CD163highHLA-DRlow monocyte subset. This could represent either a compensatory mechanism dampening the higher inflammatory condition observed with Propofol at T2 or a consequence of a higher postoperative Propofol-induced hemolysis.

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