Leukocyte Activation In Patients Research Articles

Effect of Pemafibrate on Hemorheology in Patients with Hypertriglyceridemia and Aggravated Blood Fluidity Associated with Type 2 Diabetes or Metabolic Syndrome

Persistent high serum triglyceride (TG) and free fatty acid (FFA) levels, which are common in metabolic syndrome and type 2 diabetes, are risk factors for cardiovascular events because of exacerbated hemorheology. To explore the effects of pemafibrate, a selective peroxisome proliferator-activated receptor alpha modulator, on hemorheology, we performed a single-center, nonrandomized, controlled study in patients with type 2 diabetes (HbA1c 6-10%) or metabolic syndrome, with fasting TG levels of ≥ 150 mg/dL and a whole blood transit time of > 45 s on a microarray channel flow analyzer (MCFAN). Patients were divided into a study group, receiving 0.2 mg/day of pemafibrate (n = 50) for 16 weeks, and a non-pemafibrate control group (n = 46). Blood samples were drawn 8 and 16 weeks after entry to the study to evaluate whole blood transit time as a hemorheological parameter, leukocyte activity by MCFAN, and serum FFA levels. No serious adverse events were observed in either of the groups. After 16 weeks, the pemafibrate group showed a 38.6% reduction in triglycerides and a 50.7% reduction in remnant lipoproteins. Pemafibrate treatment did not significantly improve whole blood rheology or leukocyte activity in patients with type 2 diabetes mellitus or metabolic syndrome complicated by hypertriglyceridemia and exacerbated hemorheology.

Особенности нетотической активности лейкоцитов у больных с хроническим тонзиллитом

The proportion of patients with chronic tonsillitis (CT) who visit otorhinolaryngologists in outpatient practice ranges from 4 to 35%. Increasingly, opportunistic microflora acts as a source of inflammation. An aggravating factor in the development of a chronic focus of inflammation is a change in the activity of the immune response. A few years ago, a new mechanism of behavior of human phagocytic cells in the implementation of nonspecific immunoreactivity was discovered – that is, formation of neutrophil extracellular traps (NETosis), associated with the adoption of a suicidal decision by the cells, in which chromatin is deconcentrated, and neutrophil extracellular traps (NEТs) are formed, which are released into the surrounding tissues for capturing pathogens and exposing them to high doses of antimicrobial proteins and enzymes. The significance of this immune mechanism and its role in the pathogenesis of СT remains unexplored. To date, laboratory methods for assessing the health status of patients with CT demonstrate insufficient specificity and accuracy. It remains relevant to develop a method for examining CT patients, which would allow the doctor to clearly establish the form of the disease, select the treatment, and adequately assess the health of the patient with CT during the follow-up care. Such a method, in our opinion, can be an assessment of the level of NETs in the blood and on the surface of the lacunae of the palatine tonsils (PT). 71 patients (median 38±9.86 years) were examined. The control group consisted of 54 practically healthy volunteers (median 38±10,9 years). The NETosis activity of leukocytes was evaluated in parallel in whole blood and smears from the surface of the lacunae of the PT. The data obtained indicate the undeniable diagnostic significance of the NETosis activity, considering the presence of a direct relationship with the form of CT. Therefore, the assessment of the level of NETosis can be proposed as a promising laboratory criterion for the severity of the patient’s condition, predicting the course of the disease and the effectiveness of the treatment.

Plasma extracellular DNA and neutrophilic leukocyte activity in patients with rheumatoid arthritis

Neutrophilic leukocytes play a key role for the joint damage in development of rheumatoid arthritis (RA). The specific death mode of these cells (netosis) may be an important reason of increase of cell-free DNA (cfDNA) in peripheral blood of the RA patients. Of great interest would be studies of alleged relationships between the of blood cfDNA contents being able of playing the role of an auto-antigen participating in the initiation of autoimmune reactions, and indices of neutrophil activation in this immunopathological disorder. The aim of the present study was to determine the levels of cfDNA in blood plasma of patients with RA depending on the clinical course of the disease, and to evaluate possible relationships between this index and activation of neutrophilic leukocytes. The study was conducted on 28 conditionally healthy donors and 63 patients with RA from the Rheumatology Department at the Clinic of Immunopathology (Novosibirsk). The level of cfDNA was determined using PicoGreen fluorescent dye. Neutrophils from the peripheral blood of donors and patients with rheumatoid arthritis were isolated in a Ficoll-Urografin density gradient. Neutrophilic leukocytes accounted for more than 98% of the fraction of isolated cells, and their viability was 99%. A portion of freshly isolated neutrophils was stimulated by phorbol myristate acetate. Concentration of myeloperoxidase in blood plasma of donors and patients with RA was determined using the Human MPO ELISA kit. It has been shown that the increased concentration of extracellular DNA in blood plasma of RA patients correlates with an higher degree of disease activity, and this parameter may serve as a relatively independent indicator of the disease intensity. A correlation was found between the level of cfDNA and common biochemical markers used to assess the activity of disease, i.e., DAS-28 and C-reactive protein levels in serum (p 0.05). Decrease of cfDNA concentrations is detected during treatment of the RA patients. This is due to the expected prognosis, i.e., a decreased manifestation of the disease, which also means correct administration of therapy. A relationship was found between the level of cfDNA and blood myeloperoxidase concentration in RA patients. The data obtained during the study suggest a possible connection between increased concentration of extracellular DNA, and activation of neutrophilic leukocytes in rheumatoid arthritis, with increased netosis in the affected joints.

Intrinsic and Extrinsic Cell Apoptotic Pathways in Patients with HTLV-1-Associated Myelopathy/Tropical Spastic Paraparesis: A Systematic Review.

We aimed to verify the influence of intrinsic and extrinsic cell apoptotic pathways on the inhibition of cellular apoptosis in patients with tropical spastic paralysis/myelopathy related to human T cell lymphotropic virus type 1. The databases accessed were PubMed, Scopus, Science Direct, and Web of Science. Neither the time of publishing nor the language of the articles was limited. The descriptors used for this systematic literature review were: Tropical Paraparesis, Proto-Oncogenic Protein C, Bcl-2, Bcl-X Protein, Bax protein, Fas ligand (FasL) protein, Fas receptor, TNF-related apoptosis-inducing ligand and Fas-associated protein with death domain (FADD)-like apoptosis regulating. The search resulted in 546 articles from which 9 articles were selected for analysis; ranging from serum levels of Bcl-2, Fas and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) measured by enzyme-linked immunosorbent assay and the levels of cellular expression of Bcl-2 and Bcl-xL the TCD4+ lymphocytes accessed by western blot. Most studies accessed either gene expression or polymorphism of Fas, FasL, and TRAIL in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), whereas one study used flow cytometry and fluorescence to determine Fas expression. Increased Bcl-xL expression inhibited T lymphocyte apoptosis, whereas Bcl-2, serum levels, and cellular expression did not influence T lymphocyte apoptosis and serum levels of Fas were significantly higher and associated with markers of leukocyte activation in patients with HAM/TSP. In addition, Fas polymorphism (FAS-670AA) was associated with higher proviral load. There is a need for additional research on this issue since the number of patients was small and the studies presented higher heterogeneity.

Evaluation of Pro-Inflammatory Leukocyte Activity in Patients Undergoing Cardiac Surgery

Evaluation of Pro-Inflammatory Leukocyte Activity in Patients Undergoing Cardiac Surgery

CHANGES IN THE DYNAMICS OF THE FUNCTIONAL ACTIVITY OF BLOOD GRANULOCYTES IN PATIENTS WITH PANCREATIC NECROSIS.

The purpose of research. Predicting risk of infection of acute destructive pancreatitis.
 Material and Methods. The dynamics of functional activity of granulocytes in 27 (58,7%) patients with sterile and 19 (41,3%) patients with infected pancreatic necrosis. An objective assessment of the severity of their condition carried out by performed using of the scale of the integrated assessment APPACHE-II. The exclusion criterion from the study was the revealing of accompanying acute and chronic diseases.
 Results.Scale APPACHE-II can be used mainly for screening assessment of the degree of polyorgan insufficiency, as well as predicting the development of infected pancreatic necrosis forms, but, unfortunately, only in the later stages of development of pathological process. Dynamics of changes in the functional activity of leukocytes in patients with pancreatic necrosis can be used as an additional test, which characterizes the moment of transition of sterile pancreatic necrosis to infected, and, consequently, in deciding on the timing of terms and nature of performance of operational allowance.
 Conclusions. 1. In patients with sterile pancreatic necrosis (SPN) after a moderate decrease in the activity of myeloperoxidase (MPO), the content of the cation proteins (CP) in the first three days in subsequent periods of observation, on the background of complex conservative therapy, noted, compared with the established "Normal" gradual increase or stabilization at close to the indicators of the functional activity of neutrophils. The activity of KF and alkaline phosphatase from patients with SPN, after the increase in performance in the first 11 days, then gradually declines, reaching the normal level. 2. Patients with infected pancreatic necrosis(IPN) sharply, starting with 11 days of the study, by 35-40%, reduce the activity of MPO and the content of CB and Acid Phosphatase activity and Alkaline Phosphatase, on the contrary, it had progressively pronounced growth after 11 days by 35-40%.

Platelet Serotonin Aggravates Myocardial Ischemia/Reperfusion Injury via Neutrophil Degranulation.

Platelets store large amounts of serotonin that they release during thrombus formation or acute inflammation. This facilitates hemostasis and modulates the inflammatory response. Infarct size, heart function, and inflammatory cell composition were analyzed in mouse models of myocardial reperfusion injury with genetic and pharmacological depletion of platelet serotonin. These studies were complemented by in vitro serotonin stimulation assays of platelets and leukocytes in mice and men, and by measuring plasma serotonin levels and leukocyte activation in patients with acute coronary syndrome. Platelet-derived serotonin induced neutrophil degranulation with release of myeloperoxidase and hydrogen peroxide (H2O2) and increased expression of membrane-bound leukocyte adhesion molecule CD11b, leading to enhanced inflammation in the infarct area and reduced myocardial salvage. In patients hospitalized with acute coronary syndrome, plasmatic serotonin levels correlated with CD11b expression on neutrophils and myeloperoxidase plasma levels. Long-term serotonin reuptake inhibition-reported to protect patients with depression from cardiovascular events-resulted in the depletion of platelet serotonin stores in mice. These mice displayed a reduction in neutrophil degranulation and preserved cardiac function. In line, patients with depression using serotonin reuptake inhibition, presented with suppressed levels of CD11b surface expression on neutrophils and lower myeloperoxidase levels in blood. Taken together, we identify serotonin as a potent therapeutic target in neutrophil-dependent thromboinflammation during myocardial reperfusion injury.

Release of metal ions from dental alloys and their effect on the metabolic and killing activity of leukocytes

Aim of study. We investigated the effect of metal cations solutions from dental alloys and solutions of metal salts on the functional and metabolic activity of blood leukocytes. Materials and methods. 22 patients with complaints of intolerance to dental materials (IDM) were examined. The control group consisted of 19 patients without IDM complaints. Results. After the incubation of fragments of dental alloys in water, metal cations are identified, depending on the time of incubation. This metal cations solutions (MCS) changed the metabolic and killing activity of blood neutrophils in patients with IDM, which may be a marker of intolerance to prosthetic materials. MCS and metal salts solutions (MSS) increased the secretion of IL-2, IL-4, TNF-α, INF-γ by whole blood leucocytes in patients with IDM, which indicates that hypersensitivity to the components of dental alloys was induced in the experimental group, and only IL-2, IL-10 were upregulated in the control group, which in turn indicates an immunomodulatory effect. Secretion of cytokines by blood leukocytes of patients influenced by components of dental alloys can serve as a marker of their biocompatibility and a criterion for hypersensitivity reactions. The serum level of HSP-70 in patients with IDM was 2.41 [0.79; 2.9] ng / ml, and it was lower in comparison with the control group– 4.1 [2.23; 5.11] (p <0.05). The PHA stimulated the release of HSP-70 in both groups. In patients of the experimental group MCS inhibited the secretion of HSP-70 (0.8 [0.3, 1.1] ng / ml), while in the control group it stimulated its secretion (5.7 [4.1, 8.5] ng / ml). The baseline NO-producing activity of leukocytes in patients of both groups during incubation with PHA in vitro did not differ. However, when they were cultivated with MCS, the release of NO by blood leucocytes was suppressed.

Sublingual leukocyte activation in patients with severe sepsis or septic shock

The objective of this study was to compare the number of rolling and adhered leukocytes in patients with severe sepsis/septic shock with non-infected controls. Microcirculatory flow alterations and endothelial cell dysfunction are elements of sepsis pathophysiology. Traditionally, microcirculatory emphasis has been on red blood cell vessel perfusion. However, assessment of interactions between white blood cells and endothelial cells may be another early diagnostic modality.

Glucose-dependent leukocyte activation in patients with type 2 diabetes mellitus, familial combined hyperlipidemia and healthy controls

BackgroundLeukocyte activation has been associated with vascular complications in type 2 diabetes mellitus (T2DM). Hyperglycemia may be involved in this leukocyte activation. Our aim was to investigate the role of elevated glucose concentrations on leukocyte activation in patients with a wide range of insulin sensitivity. MethodsLeukocyte activation was determined after ingestion of 75 gram glucose in subjects with T2DM, familial combined hyperlipidemia (FCH) and healthy controls. Leukocyte activation markers were measured by flow cytometry. Postprandial changes were calculated as the area under the curve (AUC), and the incremental area under the curve corrected for baseline values (dAUC). Results51 subjects (20 T2DM, 17 FCH and 14 controls) were included. Fasting neutrophil CD66b expression and CD66b-AUC were respectively 36% and 39% higher in T2DM patients than in controls (p=0.004 and p=0.003). Fasting neutrophil CD66b expression correlated positively with glucose-AUC (Spearman’s rho 0.481, p<0.001) and HbA1c (rho 0.433, p=0.002). Although fasting monocyte CD11b expression was not significantly different between subjects, monocyte CD11b-AUC was 26% higher in T2DM than in controls (p=0.006). Similar trends were observed for FCH patients. Monocyte CD11b-dAUC correlated positively with glucose-AUC (rho 0.322, p=0.022) and HbA1c (rho 0.319, p=0.023). ConclusionsThese data suggest that both acute and chronic hyperglycemia, associated with insulin resistance as seen in T2DM and FCH, are involved in the increased fasting and postprandial leukocyte activation observed in these conditions.

Postprandial glucose-dependent leukocyte activation in patients with different ranges of insulin sensitivity

Postprandial glucose-dependent leukocyte activation in patients with different ranges of insulin sensitivity

Leukocyte activity in patients with ST-segment elevation acute myocardial infarction treated with anakinra.

Anakinra, the recombinant form of the human interleukin (IL)-1 receptor antagonist, blunts the acute systemic inflammatory response in patients with ST-segment elevation myocardial infarction (STEMI), by determining a fall in peripheral blood leukocyte and plasma C-reactive protein levels. The aim of the present study was to determine the effects of anakinra on the activity of leukocytes measured ex vivo. Blood was collected 72 h after admission in 17 patients enrolled in the Virginia Commonwealth University-Anakirna Remodeling Trial (2) (VCU-ART2) and randomly treated with anakinra (N=7) or placebo (N=10). Whole blood was cultured at 37°C for 24 h to measure spontaneous production of IL-6 or stimulated with Escherichia coli lipopolysaccharide (LPS) for toll-like receptor (TLR)-4 or heat-killed Staphylococcus epidermidis (SE) for TLR-2 activation. The cultures of anakinra-treated patients produced significantly less IL-6 spontaneously (71 pg/mL [27-114]) compared with placebo-treated patients (290 pg/mL [211-617], p=0.005). LPS- or SE-induced IL-6 production, on the other hand, was not statistically different between anakinra-versus placebo-treated patients (344 pg/mL [94-560] versus 370 pg/mL [306-991], p=0.32 for LPS, and 484 pg/mL [77-612] versus 615 pg/mL [413-871], p=0.31 for SE, respectively). IL-1 blockade with anakinra in STEMI patients results in reduced spontaneous leukocyte activity ex vivo without impairing the responsiveness to bacterial stimuli.

PS13 - 4. Gender differences in glucose-dependent leukocyte activation in patients with type 2 diabetes mellitus, familial combined hyperlipidemia and healthy controls

Atherosclerosis in type 2 diabetes mellitus (T2DM) and insulin resistance (IR) is characterized by a chronic inflammatory response in which activation of leukocytes is mandatory. Acute and chronic hyperglycemia may be involved in this leukocyte activation. Female T2DM patients have more severe atherosclerosis than male diabetics and differences in inflammation may be responsible for this observation.

Bivalirudin reduces platelet and monocyte activation after elective percutaneous coronary intervention

Concomitant antithrombotic therapy is essential for the prevention of ischaemic events in percutaneous coronary intervention (PCI) and stenting. With new anticoagulant medications being developed and applied in PCI, this raises the question of possible interactions with platelet and leukocyte activation. We therefore sought to investigate the influence of bivalirudin and heparin in platelet and leukocyte activation in patients undergoing elective PCI. Forty-six patients were recruited consecutively in the setting of the Intracoronary Stenting and Antithrombotic Regimen-Rapid Early Action for Coronary Treatment (ISAR-REACT)-3 trial and were randomly assigned to receive either unfactionated heparin or bivalirudin during elective PCI. Surface expression of CD62P (P-Selectin), CD42b (GPIbalpha), CD40L, PAC-1 on circulating platelets and CD11b, CD14 and CD15 on circulating leukocytes were evaluated by flow cytometry. Cytokine levels of IL-12p70, tumour necrosis factor (TNF), IL-8, IL-6, IL-1beta and IL-10 were determined by cytometric bead array. Platelet surface expression of PAC-1, P-Selectin and GPIbalpha was significantly reduced after PCI in patients receiving bivalirudin as compared to heparin. Similarly, CD11b expression on CD14+ monocytes was diminished after bivalirudin. However, no differences were observed in cytokine levels between the bivalirudin and the heparin group, before or after PCI. In conclusion, our data suggest that bivalirudin may reduce platelet and monocyte activation in patients undergoing elective PCI. Thereby, bivalirudin might reduce periinterventional thrombotic complications.

CELLULAR IMMUNITY INDEXES AND THEIR CHANGES BY INFLUENCE OF SOLUBLE STAPHYLOCOCCUS AUREUS PRODUCTS IN PATIENTS WITH PURULENT INFLAMMATORY DISEASES

Receptor and functional features of peripheral blood leukocytes in recurrent purulent inflammatory patients depending on etiology and clinical factors and influence in vitro of soluble Staphylococcus aureus products on leukocytes function were examined. A dependence of cellular immunity indexes on etiology of process was determined. It was showed that staphylococcal culture cell-free supernates stimulate in vitro phagocytosis and oxidative burst activity of leukocytes in patients who has disorders caused by Staphylococcus aureus.

Elevation of Endothelial Microparticles, Platelets, and Leukocyte Activation in Patients With Venous Thromboembolism

Elevation of Endothelial Microparticles, Platelets, and Leukocyte Activation in Patients With Venous Thromboembolism

ENDOTHELIAL MICROPARTICLE BINDING TO LEUKOCYTES AS A DETERMINANT OF LEUKOCYTE ACTIVATION AND NITRIC OXIDE EXPRESSION IN ACUTE VENOUS THROMBOEMBOLISM

PURPOSE: The pathophysiology of venous thromboembolism (VTE) involves complex cellular interactions resulting from endothelial damage, hypercoagulability and inflammation. Increased levels of endothelial microparticles (EMP) and leukocyte activation have been reported in VTE. EMP have been shown to induce monocyte and neutrophil activation in vitro. Whether this phenomenon determines leukocyte activation in vivo is unknown. The purpose of this study is to determine the correlation between the leukocyte-bound EMP and leukocyte activation in patients with VTE.

Endothelial Microparticles and Platelet and Leukocyte Activation in Patients With the Metabolic Syndrome

Accumulating evidence has shown a strong association between the metabolic syndrome (MS) and a chronic inflammatory state predisposing to atherosclerosis. We investigated leukocyte, platelet, and endothelial activation markers and cellular interactions in 33 patients with the MS and 25 healthy controls. Using flow cytometry, we measured: (1)P-selectin expression in platelets; (2) platelet microparticles identified by CD31 expression; (3) endothelial microparticles (EMPs) identified by expression of CD31 (EMP(31)), CD62E (EMP(62E)), and CD51 (EMP(51)); (4) conjugates of leukocytes with platelet microparticles/platelets and with EMPs identified by CD54 (EMP(54)); and (5) CD11b expression in leukocytes. Patients with the MS had markedly elevated EMP(31), although EMP(62E) levels were normal, suggesting that EMP(31) levels were increased because of endothelial cell apoptosis, rather than activation. EMP(51), EMP(54)-lymphocyte conjugates, platelet expression of P-selectin, CD11b expression in leukocytes, and platelet-lymphocyte conjugates were also increased in patients with the MS. Platelet-leukocyte conjugates correlated with leukocyte activation, suggesting that platelet binding to leukocytes regulates leukocyte activation in vivo. In conclusion, our data demonstrate endothelial cell microparticle release, platelet and leukocyte activation, and increased binding of EMPs and platelets to leukocytes in patients with the MS.

Elevation of Endothelial Microparticles, Platelets, and Leukocyte Activation in Patients With Venous Thromboembolism

The purpose of this research was to determine the levels of platelet, leukocyte, and endothelial activation and markers of cellular interactions in patients with venous thromboembolism (VTE). The details of interactions between endothelium, platelets, and leukocytes in VTE are not well understood. We studied 25 patients with VTE and compared 25 healthy controls. We used flow cytometry to measure: 1) endothelial microparticles (EMP) identified by CD31+/CD42b- (EMP(31)) or E-selectin (EMP(62E)); 2) platelet microparticles (CD31+/CD42b+); 3) surface expression of P-selectin in platelets and CD11b in leukocytes; 4) EMP-monocyte conjugates (percentage of monocytes positive for E-selectin); and 5) platelet-leukocyte conjugates (PLC) expressed as percentage of leukocytes positive for CD41. Patients with VTE had marked elevations of EMP(31) (2,193 vs. 383 counts/microl; p = 0.003), EMP(62E) (368 vs. 223 counts/microl; p = 0.001), and EMP-monocyte conjugates (3.3% vs. 2.5%; p = 0.002), as well as increased activation of platelets (35.2 vs. 5.0 fluorescence intensity units for P-selectin; p < 0.0001) and leukocytes (13.9 vs. 7.7 U for CD11b; p = 0.004). Also elevated in VTE were PLC (61.7% vs. 39.6%; p = 0.01). Expression of CD11b in leukocytes strongly correlated with PLC (r = 0.74; p < 0.0001). Marked activation of endothelium, platelets, and leukocytes occurs in VTE, and VTE, or the accompanying inflammatory process, involves the release of EMP and formation of EMP-monocyte conjugates and PLC. These findings support prior studies suggesting that release of EMP and their binding to monocytes are key events in thrombogenesis. Our findings also support the concept that the formation of PLC regulates leukocyte activation and participates in linking thrombosis with inflammation.

Systemic leukocyte activation in patients with central giant cell lesions

Central giant cell lesion (CGCL) is a reactive lesion of the jaws with an associated inflammatory infiltrate. Since cell circulation allows for intense communication between different compartments in the body, we investigated whether the CGCL would lead to phenotypic and/or functional changes in circulating leukocytes. We obtained lymphocytes and monocytes from CGCL patients and control subjects, to evaluate cytokine and adhesion molecule expression using flow cytometry. Our results revealed that CD4(+) T cells and CD14(+) monocytes from CGCL express elevated levels of CD11a and CD11b, respectively, when compared with controls. The frequencies of CD4(+) T cells expressing interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha and the frequencies of CD4(+) and CD14(+) cells expressing interleukin (IL)-10 were increased in CGCL group, when compared with controls. Our data indicate that, although CGCL is a localized lesion, the patients show systemic functional alterations in circulating leukocytes, suggesting their role in the inflammatory pathogenesis of CGCL.