Manuela Kudlova

The long pentraxin 3 in cardiac surgery: distinct responses in "on-pump" and "off-pump" patients.

Objective. Pentraxin 3 (PTX3) is a newly identified acute phase reactant with non-redundant functions in innate immunity. The purpose of this study was to assess the kinetics of release of PTX3 in cardiac surgical patients, operated on either with or without the use of cardiopulmonary bypass (CPB). Design. Thirty-four patients, seventeen in each group, were randomly assigned to CABG surgery performed either with (“on-pump”) or without (“off-pump”) CPB. Blood samples were collected both during and after the operation up to the 7th day. Results. In patients operated on with the use of CPB, PTX3 levels increased throughout the operation. Compared to baseline levels the highest PTX3 value (p<0.000) was attained on the 1st postoperative day in both “on-pump” and “off-pump” patients. In contrast to CPB patients, however, PTX3 levels in the latter group declined slowly, remaining elevated as long as the 3rd postoperative day (p<0.042). Conclusions. Operations performed with the use of CPB are associated with a more pronounced release of PTX3 immediatelly after operation.

Early Expression of FcRI (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, FcγRI (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 FcγR1 (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.

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 163TM 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 1st 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 1st postoperative day, being little influenced by cardiopulmonary bypass.

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 μg/mL; 37.99±16.58 μg/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.

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.

J774 macrophage-like cell line cytokine and chemokine patterns are modulated by Francisella tularensis LVS strain infection

Mutual interactions were investigated between intracellular parasitic bacterium Francisella tularensis (F.t.; highly virulent bacterium responsible for tularemia, replicating within the host macrophages) and murine macrophage-like cell line J774. Recombinant murine lymphokine INF-γ and/or LPS derived from E. coli were determined to stimulate in vitro antimicrobial activity of macrophage-like J774 cell line against the live vaccine strain (LVS) of F.t. through their ability to produce proinflammatory cytokines and chemokines. F.t. infection up-regulated IL-12 p40 production and down-regulated TNF-α production by stimulated macrophages; on the other hand, F.t. infection did not affect the production of IL-8, IL-6, MCP-5, and RANTES by stimulated macrophages. This showed that F.t. infection modulates the cytokine synthesis by J774 macrophage cell line.

The Effect of Interferon-γ and Lipopolysaccharide on the Growth of Francisella tularensis LVS in Murine Macrophage-like Cell Line J774

BACKGROUND Francisella tularensis, a causative agent of human tularemia, displaying the ability to proliferate inside the human cells. AIMS To evaluate the growth potential of F. tularensis LVS strain in macrophage-like cell line J774 modulated by recombinant interferon γ and E. coli derived lipopolysaccharide. RESULTS Stimulation of J774 cells either by interferon-γ or lipopolysaccharide alone, or especially in combination before infection F. tularensis, revealed protective effects. Higher concentrations of stimulating agents were needed to inhibit ongoing F. tularensis infection. CONCLUSIONS Stimulation of J774 cell line by combination of interferon-γ with lipopolysaccharide inhibits the intracellular growth of F. tularensis.

The inflammatory response in cardiac surgery. An up-to-date overview with the emphasis on the role of heat shock proteins (HSPs) 60 and 70.

Cardiac surgical operations are associated with the development of a systemic inflammatory response syndrome (SIRS). In most cases, the inflammatory response is apparent only in its subclinical form. In some cases, however, it can become clinically manifest or even life-threatening. This review article presents some new data regarding its biological importance, along with an insight into the recently discovered role played by heat shock proteins 60 and 70.