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

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.

Pentraxin 3 and other inflammatory biomarkers related to atrial fibrillation in cardiac surgery.

OBJECTIVES: The aim was to evaluate the association between perioperative inflammatory biomarkers and atrial fibrillation (AF) in cardiac surgical patients. METHODS: Forty-two patients undergoing cardiac surgery were divided into three groups according to the occurrence of AF: Group A (n = 22) - patients with no AF, Group B (n = 11) - patients with new onset AF postoperatively and Group C (n = 9) - patients with preoperative history of atrial fibrillation. The serum levels of PTX3, CRP, TLR2, IL-8, IL-18, sFas, MMP-7 and MMP-8 were measured at the following time points: before surgery, immediately and 6 h after surgery and on the 1st, 3rd and 7th postoperative days (POD). RESULTS: Serum levels of PTX3 showed a significant difference between Groups A and C on the 3rd POD (p<0.05) and on the 7th POD (p<0.0001). IL-8 levels were different between Groups A and C immediately after surgery (p<0.05), 6 hours after surgery (p<0.05) and on the 3rd POD (p<0.05). There was a difference between Groups B and C on the 1st POD in IL-8 levels (p<0.05). The sFas levels differed between Groups A and C on the 3rd POD (p<0.01) and the 7th POD (p<0.05). There was also a difference on the 7th POD (p<0.05) between the Groups B and C. No significant differences between the groups was seen for other biomarkers. CONCLUSION: This study demonstrates significantly different dynamics of PTX3, IL-8 and sFas levels after cardiac surgery in relation to AF.

Impact of cardiac surgery on the expression of CD40, CD80, CD86 and HLA-DR on B cells and monocytes.

OBJECTIVE: We measured and compared changes in the percentage of cells expressing CD80, CD86, CD40, HLA-DR and the expression of these molecules on B cells and monocytes of patients who underwent either on-pump, mini on-pump or off-pump cardiac surgery. METHODS: Blood samples from patients who underwent either on-pump, mini on-pump or off-pump cardiac surgery were collected before surgery, instantly after surgery and on the 1(st), 3(rd) and 7(th) days after surgery. Surface expression of CD80, CD86, CD40 and HLA-DR molecules was determined by flow cytometry. RESULTS: Our results show that all three surgical techniques altered the expression of these molecules, as well as the percentage relative number of specific cell populations. We identified statistically significant differences when comparing different surgical techniques. On-pump surgery revealed a more pronounced impact on the phenotype of immune system cells than the other techniques. Therefore, it is likely that the function of immune cells is changed the most by on-pump surgery. We found a lower decrease in the number of CD80(+) monocytes and a lower drop in the CD40 expression on monocytes in off-pump patients in comparison with on-pump patients. CONCLUSION: All the types of cardiac surgical techniques, off-pump, on-pump and modified mini-invasive on-pump, are associated with changes in CD80, CD86, CD40 and HLA-DR expression. We found several significant differences in the expression of the selected molecules when we compared all three groups of patients.

Could pentraxin 3 be a new diagnostic marker for excessive inflammatory response in cardiac surgery?

BACKGROUND: The aim of this study was to compare the dynamics of two inflammatory response biomarkers pentraxin 3 (PTX3) and C-reactive protein (CRP) after cardiac surgery with particular regard to different postoperative clinical manifestation of inflammatory response. PATIENTS AND METHODS: In this study, 42 patients undergoing open heart surgery with the use of cardiopulmonary bypass were included and divided in two groups according to the extent of clinical manifestation of inflammatory response: Group A (n=21)-patients with different severity of systemic inflammatory response syndrome (SIRS) and Group B (n=21)-patients with uneventful postoperative period (no SIRS). The serum levels of PTX3 and CRP were evaluated and compared at the following time points: before and at the end of surgery, 6 hours, 1st, 3rd, and 7th day after surgery. RESULTS: The dynamics of CRP levels were comparable between both groups and showed the classical characteristics after cardiac surgery with a peak on the 3rd postoperative day (113 vs. 132 mg/L). In contrast, the dynamics of PTX3 showed an earlier increase of serum levels with the peak on the 1st postoperative day in both groups (36.3 vs. 42.7 ng/mL). Importantly, a significant difference of PTX3 levels was found on the 3rd postoperative day (31.1 vs. 7.0 ng/mL; p<0.006) between the two groups showing significantly delayed decrease of PTX3 levels in patients with SIRS (Group A). CONCLUSION: This study demonstrates considerably different dynamics of PTX3 levels after cardiac surgery in patients with SIRS and patients without SIRS, thus it may be indicative to start the appropriate therapy.

TLR2 and TLR4 expression on blood monocytes and granulocytes of cardiac surgical patients is not affected by the use of cardiopulmonary bypass.

Cardiac surgery is inseparably linked to the activation of innate immunity cells recognizing danger signals of both endogenous and exogenous origin via pattern recognition receptors such as TLR receptors. Therefore, we followed by flow cytometry TLR2 and TLR4 expression on blood monocytes and granulocytes of patients who underwent coronary artery bypass grafting using beating heart surgery (off-pump, n = 34), with use of standard cardiopulmonary bypass (CPB), (on-pump, n = 30), and miniinvasive CPB (mini on-pump, n = 25), respectively, before, during surgery, and up to 7th postoperative day. TLR2 and TLR4 expression both on monocytes and granulocytes was significantly diminished already at the end of CPB being highly significantly decreased at the end of surgery in all patients' groups. TLR2 and TLR4 expression reached preoperative value at the 1st postoperative day being significantly higher at the 3rd postoperative day. Using intracellular staining we found the peak of TLR2 and TLR4 expression inside of monocytes and granulocytes at the first postoperative day in a subgroup of on-pump patients. In conclusion, TLR2 and TLR4 expression is significantly modulated in patients undergoing coronary artery bypass grafting as a part of adaptive homeostatic mechanisms induced by major surgery. The very surgical trauma is responsible for TLR2 and TLR4 modulation. Surprisingly, cardiopulmonary bypass itself was little contributing to the modulation of TLR2 and TLR4 expression.

The effect of conventional and mini-invasive cardiopulmonary bypass on neutrophil activation in patients undergoing coronary artery bypass grafting.

Interleukin-10 (IL-10) is considered to be a cytokine with potent anti-inflammatory properties, which have been previously linked to increased incidence of sepsis. The level of IL-10 is elevated by cardiac surgery when cardiopulmonary bypass (CPB) and methylprednisolone are used. In our study, we compare the level of IL-10, IL-10 Receptor (IL-10R), and percentage of neutrophils between two groups of cardiac surgical patients undergoing Coronary Artery Bypass Grafting, both of which were not given methylprednisolone. The first group was operated with conventional CPB, while the second group was operated with minimally invasive CPB (mini-CPB). We detected enhanced level of IL-10 during surgery and at the end of surgery in both groups of patients. While no correlation between IL-10 and IL10R was found, IL-10 was positively correlated with increased percentage of neutrophils at the time points when the level of IL-10 peaked.

CD200/CD200R paired potent inhibitory molecules regulating immune and inflammatory responses; Part II: CD200/CD200R potential clinical applications.

SUMMARY: CD200 and its receptor were recognized as having the multiple immunoregulatory functions. Their immunoregulatory, suppressive, and tolerogenic potentials could be very effectively exploited in the treatment of many diseases, e.g. Alzheimer disease, rheumatoid arthritis, and allergy to name only some. Many research projects are aimed to develop clinically valuable methods being based on the structure and function of these paired molecules. In this review, we would like to introduce CD200/CD200R functions in a clinical context.

CD200/CD200R paired potent inhibitory molecules regulating immune and inflammatory responses; Part I: CD200/CD200R structure, activation, and function.

CD200/CD200R are highly conserved type I paired membrane glycoproteins that belong to the Ig superfamily containing a two immunoglobulin-like domain (V, C). CD200 is broadly distributed in a variety of cell types, whereas CD200R is primarily expressed in myeloid and lymphoid cells. They fulfill multiple functions in regulating inflammation. The interaction between CD200/CD200R results in activation of the intracellular inhibitory pathway with RasGAP recruitment and thus contributes to effector cell inhibition. It was confirmed that the CD200R activation stimulates the differentiation ofT cells to the Treg subset, upregulates indoleamine 2,3-dioxygenase activity, modulates cytokine environment from a Thl to a Th2 pattern, and facilitates an antiinflammatory IL-10 and TGF-beta synthesis. CD200/CD200R are required for maintaining self-tolerance. Many studies have demonstrated the importance of CD200 in controlling autoimmunity, inflammation, the development and spread of cancer, hypersensitivity, and spontaneous fetal loss.

Expression of urokinase plasminogen activator receptor on monocytes and granulocytes is modulated by cardiac surgery.

AIMS: Coronary artery bypass grafting (CABG) is linked to the induction of the blood coagulation/fibrinolysis cascade, which is an integral component of inflammation induced by cardiac surgery. We followed the modulation of urokinase plasminogen activator receptor uPAR (CD87) separately for monocytes and granulocytes in blood of cardiac surgery patients. METHODS: Expression of uPAR, analyzed as Median Fluorescence Intensity (MFI), on blood monocytes and granulocytes was determined by flow cytometry. Changes in uPAR expression in patients undergoing CABG using standard cardiopulmonary bypass ("on-pump") were compared to the changes in uPAR expression in patients undergoing CABG using mini-invasive cardiopulmonary bypass ("mini on-pump"). RESULTS: In "on-pump" patients, the median of uPAR expression on granulocytes before surgery was 18.1 (InterQuartile Range (IQR): 15.6-20.4). uPAR expression was significantly decreased after surgery (p<0.001), on the first postoperative day (p<0.001), and on the third postoperative day (p<0.05). In "mini on-pump" patients, the median of uPAR expression on granulocytes before surgery was 15.2 (IRQ: 13.8-19.4). The significantly decreased uPAR expression was found only at the end of surgery (p<0.05). The similar pattern of uPAR expression was also found for monocytes. The preoperative level in "on-pump" patients was 23.3 (IRQ: 18.9-30.2). There was significantly decreased uPAR expression at the end of surgery (p<0.01) and at the first postoperative day (p<0.05). In "mini on-pump" patients, the preoperative uPAR expression was 16.9 (IQR: 14.5-20.2). Expression of uPAR was significantly decreased only after surgery (p<0.05). When comparing "onpump" patients to "mini on-pump" patients, no significant differences in the expression of uPAR were found. CONCLUSION: uPAR expression on granulocytes and monocytes is significantly modulated by cardiac surgery.

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.

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, Q(3)=64) to a median=88, (Q(1)=62, Q( 3)=109.5; p<0.01) at the 3(rd) postoperative day and median=74, (Q(1)=63, Q(3)=84.5; p<0.01) at the 7(th) postoperative day. In "off-pump" patients, granulocyte CD95 MFI was median=55, (Q(1)=51, Q(3)=84) before surgery. The significant increase was found on the 3(rd) postoperative day only; median=90, (Q( 1)=66; Q(3)=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 3(rd) postoperative day (p<0.05) and at the 7(th) 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.

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

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.

The counter-regulation of atherogenesis: a role for interleukin-33.

The recently recognized cytokine interleukin-33 and its receptor ST2 play a favorable role during atherogenesis by inducing a Th1 --> Th2 shift of the immune response. IL-33 also protects the failing human heart from harmful biomechanical forces which lead to cardiomyocyte hypertrophy and exaggerated interstitial fibrosis. IL-33 inevitably displays side effects common to other Th2 cytokines, the most grave of which is a predisposition to allergic reactions. IL-33 is a nuclear transcription factor of endothelial cells. As such, it is abundant in nonproliferating vessels. Its down-regulation is required for angiogenesis, which may be profitable in wound healing or deleterious in tumor growth.

Serum level of sCD163, a soluble receptor for hemoglobin, is influenced by cardiac surgery.

BACKGROUND: The scavenger receptor for complexes hemoglobin-haptoglobin (CD163), which is expressed on monocytes/ macrophages, is shed to the body fluids in a soluble form (sCD163). OBJECTIVES: To evaluate the dynamics of sCD163 in the blood of patients undergoing cardiac surgery. PATIENTS AND METHODS: Sixty-one adult patients who underwent coronary artery bypass grafting (CABG) were enrolled in the study. They were assigned to undergo CABG using either cardiopulmonary bypass (CPB), "on-pump", (22 patients), modified CPB, mini "on-pump", (17 patients) or without CPB, "off-pump", (22 patients) surgery. Serum levels of sCD163 in venous blood samples taken before and after surgery, and during an early postoperative period, were evaluated by Macro 163(TM) diagnostic kit (IQ Products, Groningen, NL). RESULTS: Compared to the preoperative levels ("on-pump"; 344 ng/mL, "off-pump"; 314.5 ng/mL, mini-invasive "on-pump"; 336.5 ng/mL) serum levels were elevated at the finish of surgery, reaching maximum at the 1(st) postoperative day ("onpump"; 658 ng/mL; p<0.05, "off-pump"; 810.5 ng/mL; p<0.01; mini-invasive "on-pump"; 663 ng/mL; non-significant).No significant differences regarding the serum levels of sCD163 between different surgical approaches were found. CONCLUSION: Serum level of sCD163 scavenger molecule for hemoglobin is elevated at the end of surgery and at the 1(st) postoperative day, being little influenced by cardiopulmonary bypass.

Occurrence and significance of the nuclear transcription factor Krüppel-like factor 4 (KLF4) in the vessel wall.

Practically all mammalian cells including human can switch, according to micro- or macroenvironmental conditions, from states of cellular quiescence to inflammatory activation and vice versa. Along with recent knowledge, cellular quiescence is not a passive, but a highly active state with broad engagement of the cell synthetic and secretory machinery. Inflammatory activation is a beneficial process in cases of infection; however, if its control fails, it may degrade into autoimmune diseases or cancer growth. Control over cellular quiescence is exerted predominantly by a set of zinc-finger transcription proteins, referred to as Krüppel-like factors (KLFs). This review article offers recent information concerning activities of Krüppel-like factor 4 in the vascular wall.

RANK/RANKL expression is induced by cardiac surgical operation.

BACKGROUND: Cardiac surgery provokes a systemic inflammatory response in any patient. This complex body reaction involves also RANK/RANKL molecules which have been recently identified as principal regulators of bone metabolism. AIMS: To follow the changes in the expression of RANK/RANKL molecules on innate immune cells of cardiac surgical patients. PATIENTS AND METHODS: Twenty-six patients undergoing cardiac surgical were assigned to undergo coronary artery bypass grafting using either cardiopulmonary bypass ("on-pump") or modified "miniinvasive on-pump". The expression of RANK/RANKL was performed by flow cytometry. RESULTS: Significantly increased expression of RANK on monocytes of "miniinvasive on-pump" patients was found at the 1st, the 3nd, and 7th postoperative days. The similar pattern was found also for monocyte RANKL expression. In addition, RANKL expression was significantly increased at the 3rd postoperative day in "on-pump" patient. No significant differences between "miniinvasive on-pump" and "on-pump" cardiac surgical patients were found. CONCLUSION: The expression of both RANK and RANKL molecules is significantly enhanced on monocytes of "miniinvasive on-pump" cardiac surgical patients.

Expression of Toll-like receptors 2 and 4 on innate immunity cells modulated by cardiac surgical operation.

OBJECTIVE: This study aimed to follow-up on the changes in the expression of Toll-like receptors (TLRs) on monocytes and granulocytes in venous blood of patients undergoing cardiac surgical operation. MATERIAL AND METHODS: TLR2 and TLR4 expression on blood cells was determined by flow cytometry in 40 patients undergoing cardiac surgery performed either with cardiopulmonary bypass (CPB) ("on-pump") or without it ("off-pump"). RESULTS: Intensity of the expression of TLR2 on both monocytes and granulocytes, expressed as median fluorescence intensity, is significantly reduced during CPB, being lower in both groups at the finish of surgery. These changes are not so remarkable in the case of TLR4 expression. Compared to "on-pump" patients, there is a higher relative number of TLR2(+) granulocytes in "off-pump" patients at the finish of surgery and of TLR4(+) granulocytes on the first postoperative day. CONCLUSIONS: We found characteristic patterns in the expression of TLR2 and TLR4 on monocytes and granulocytes in venous blood of patients undergoing cardiac surgery with or without CPB.

Early expression of FcgammaRI (CD64) on monocytes of cardiac surgical patients and higher density of monocyte anti-inflammatory scavenger CD163 receptor in "on-pump" patients.

OBJECTIVE: Activation of innate immunity cells is inseparably linked to cardiac surgical operation. The aim of this study was to assess the kinetics in the expression of receptor for Fc part of IgG, FcgammaRI (CD64), and scavenger receptor CD163 on peripheral blood cells of cardiac surgical patients and to examine the effect of cardiac bypass as a separable influence on the systemic acute inflammatory response. METHODS: Forty patients, twenty in each group, were randomly assigned to CABG surgery performed either with "on-pump" or without "off-pump" cardiopulmonary bypass. Standardized quantitative flow cytometry method was used to determine the expression of surface markers. RESULTS: The density of CD64 molecule on monocytes reached maximum on the 1st postoperative day (P<.001) whereas the peak for CD64 molecule expression on granulocytes was postponed to the 3rd postoperative day (P<.001). The expression of CD163 scavenger molecule on monocytes reached maximum on the 1st postoperative day (P<.001). The density of CD163 molecule on monocytes on the 1st postoperative day is significantly higher in "on-pump" patients in comparison with "off-pump" patients (P<.001). CONCLUSION: In cardiac surgical patients the expression of activation marker FcgammaR1 (CD64) on monocytes is increased earlier in comparison with granulocytes in both "on-pump" and "off-pump" patients. The expression of scavenger molecule CD163 on monocytes is significantly higher in "on-pump" patients.

The effect of cardiac surgery on peripheral blood lymphocyte populations.

BACKGROUND: Cardiac surgery using cardiopulmonary bypass (CPB) is associated with some adverse postoperative complications caused by an altered immune response. An alternative approach to cardiac surgery, operating without the use of CPB (i.e. off-pump surgery), seems to display less adverse impacts on the immune response. PATIENTS AND METHODS: Peripheral blood lymphocytes in 40 patients undergoing cardiac surgery either with CPB ("on-pump") or without CPB ("off-pump") were followed using flow cytometry. The samples of peripheral blood were taken at five intervals: preoperatively, after termination of the surgery, on the first, on the third and on the seventh postoperative day, respectively. RESULTS: The most substantial changes appeared on the first postoperative day in both subgroups of patients. While the percentage of both total T cells and CD4+ T cells were decreased, the percentage of HLA-DR+ activated lymphocytes was increased. These changes were more profound in the "on-pump" subgroup compared to the "off-pump" subgroup. CONCLUSION: Our results may suggest that the "off-pump" surgical approach reveals less adverse impact on adaptive immune responses.

Lipopolysaccharide binding protein and sCD14 are not produced as acute phase proteins in cardiac surgery.

OBJECTIVES: The changes in the serum levels of lipopolysaccharide binding protein (LBP) and sCD14 during cardiac surgery were followed in this study. DESIGN: Thirty-four patients, 17 in each group, were randomly assigned to coronary artery bypass grafting surgery performed either with ("on-pump") or without ("off-pump") cardiopulmonary bypass. LBP and sCD14 were evaluated by ELISA. RESULTS: The serum levels of LBP were gradually increased from the 1st postoperative day and reached their maximum on the 3rd postoperative day in both "on-pump" and "off-pump" patients (30.33+/-9.96 microg/mL; 37.99+/-16.58 microg/mL), respectively. There were no significant differences between "on-pump" and "off-pump" patients regarding LBP. The significantly increased levels of sCD14 from the 1st up to the 7th postoperative day in both "on-pump" and "off-pump" patients were found with no significant differences between these groups. No correlations between LBP and sCD14 and IL-6, CRP and long pentraxin PTX3 levels were found. CONCLUSIONS: The levels of LBP and sCD14 are elevated in cardiac surgical patients being similar in both groups. These molecules are not produced as acute phase proteins in these patients.

Essential PTX3 biology (not only) for cardiologists and cardiac surgeons.

Inflammation has been recognized to form an integral part of the atherosclerotic process. Much consideration has been given lately to the role played in atherogenesis by C-reactive protein (CRP). Although not accepted unequivocally, CRP appears to be not only a marker, but also an active mediator of the atherosclerotic process. Pentraxin 3 (PTX3) is a newly identified acute phase reactant which shares some structural and some functional properties with CRP. On the other hand, pentraxin 3 displays unique biological properties of its own, including a possible role in the pathogenesis of cardiovascular diseases and in processes accompanying the natural evolution of surgical wounds. This review article discusses recent information concerning basic pentraxin 3 biology in inflammation and in innate immunity reactions as viewed by a cardiologist in the context of acute coronary events and by a surgeon in patients struck with multiple wounds who are at the same time menaced by bacterial infections.