Neutrophil Activation In Patients Research Articles

Functional activity of blood neutrophiles in acute inflammatory diseases of the abdominal organs and abdominal tuberculosis

The diagnosis of various pathomorphological forms of acute appendicitis (AA), acute mesadenitis (AM), and abdominal tuberculosis (AT) remains an urgent problem in medicine. Neutrophilic granulocytes are the first link in the nonspecific immune response to inflammation, which is mediated by phagocytosis and intracellular bactericidal systems. Therefore, the research aimed to establish the features of the functional state of neutrophilic granulocytes in the blood of patients with AA, AM, and AT.Materials and methods. 30 practically healthy, 27 patients with AM, 40 patients with acute phlegmonous appendicitis were examined; 20 patients with acute gangrenous appendicitis, 30 patients with AT. The phagocytic activity of neutrophils in the test with latex granules, redox activity (spontaneous HCT test), cationic lysosomal proteins in the cytochemical test with bromine phenol blue was determined in the blood.Results. Phagocytic activity of neutrophils in patients with destructive forms of AA and AT was lower, and in the group of patients with AM – did not differ from control values. The phagocytic index of neutrophils in patients with destructive forms of AA was 1.5 times lower (p<0.05), and the phagocytic number in patients with gangrenous AA was 1.7 times lower than in healthy individuals (p<0.05). In patients with AM, the percentage of HCT-positive neutrophils was 1.3 times higher, in patients with destructive forms of AA and AT – 2 times higher than in healthy groups (p<0.05). The number of neutrophils containing cationic lysosomal proteins was also higher in destructive forms of AA, and in AM and AT - did not differ from the value in the control group.Conclusions. It has been established that peripheral blood neutrophils in conditions of acute destructive inflammation and abdominal tuberculosis have reduced absorption capacity with simultaneous significant activation of redox processes. Cytochemical tests to detect the phagocytic and metabolic activity of blood neutrophils are available and highly informative for the diagnosis and prediction of inflammation, as well as for the differential diagnosis of bacterial and viral diseases

Association of Neutrophil Activation, More Than Platelet Activation, With Thrombotic Complications in Coronavirus Disease 2019.

BackgroundSARS-CoV-2 infection is associated with hypercoagulability which predisposes to venous thromboembolism (VTE).We analyzed platelet and neutrophil activation in COVID-19 patients and their association with VTE.MethodsHospitalized COVID-19 patients and age- and sex-matched healthy controls were studied. Platelet and leukocyte activation, neutrophil extracellular traps (NETs), and matrix metalloproteinase-9 (MMP-9), a neutrophil-released enzyme, were measured. Four patients were re-studied after recovery. The activating effect of COVID-19 plasma on control platelets and leukocytes and the inhibiting activity of common antithrombotic agents on it were studied.Results36 COVID-19 patients and 31 healthy controls were studied; 8/36 COVID-19 patients (22.2%) developed VTE. Platelets and neutrophils were activated in COVID-19 patients. NET, but not platelet activation, biomarkers correlated with disease severity and were associated with thrombosis. Plasmatic MMP-9 was significantly increased in COVID-19 patients.Platelet and neutrophil activation markers, but less so NETs, normalized after recovery.In vitro, plasma from COVID-19 patients triggered platelet and neutrophil activation and NET formation, the latter blocked by therapeutic dose low-molecular weight heparin, but not by aspirin or dypiridamole.ConclusionsPlatelet and neutrophil activation are key features of COVID-19 patients. NET biomarkers may help to predict clinical worsening and VTE, and may guide LMWH-treatment intensity.

CHARACTERISTICS OF THE DYNAMICS OF LIPID PEROXIDATION, ANTIOXIDANT PROTECTION, AND PHAGOCYTIC ACTIVITY OF NEUTROPHILS IN THE TREATMENT OF ACUTE ODONTOGENIC OSTEOMYELITIS OF THE JAW

At the present stage, odontogenic osteomyelitis is one of the common purulent-inflammatory pathologies of the maxillofacial region. There is a tendency to change the nature of the clinical course of the disease: there is a predominance of a sluggish process with a tendency to chronicity and the development of complications. An unexpressed picture of the diagnosis of acute odontogenic osteomyelitis is difficult, which explains the delay in seeking help. The aim of the study was to determine and compare indicators of lipid peroxidation, antioxidant defense and phagocytic activity of neutrophils in patients with acute odontogenic osteomyelitis who received standard and combination treatment with Melaxen. The study was conducted on 30 patients with an established diagnosis of the Department of Maxillofacial Surgery of the Republican Clinical Hospital in Vladikavkaz. The indicators of lipid peroxidation, antioxidant protection, and phagocytic activity of neutrophils in patients with acute odontogenic osteomyelitis who received standard treatment and combined with the use of the drug “Melaxen” were evaluated. It was found that the disease can cause an increase in the concentration of malondialdehyde and hydroperoxides and a decrease in catalase activity. Under the influence of pathology, the absorption and digestion functions of neutrophils are inhibited. The efficacy of the use of the drug "Melaxen" in the treatment of acute odontogenic osteomyelitis due to its ability to suppress lipid peroxidation processes, restore antioxidant defense and phagocytic activity of neutrophils has been proven. the inclusion of Melaxen in the standard treatment regimen allows for faster reduction of oxidative stress and restoration of antioxidant protection, and also helps to normalize the phagocytic activity of neutrophils in a shorter time.

Analysis of neutrophils functional activity in men with mechanical jaundice of various genesis.

Obstructive jaundice (OJ) or blockage of the bile duct code K83.1 (according to ICD 10), occurs in approximately 45-50% of cases of all varieties of jaundice, it can be both non-tumor and tumor genesis. The functional pathway plays a special role in the genesis of complications of breast the activity of neutrophils as key effector cells responsible for the development of the inflammatory process in the breast. Investigation of the metabolic mechanisms of the functioning of neutrophils allows us to identify intracellular targets, when exposed to It was possible to modulate the level of cell reactivity.The study used data from 47 men with obstructive jaundice of non-tumor origin and 45 men with obstructive jaundice of tumor origin (stage I-II of the tumor process). As a control, data from 100 practically healthy men were used. A pronounced change in the kinetics of the chemiluminescent response of neutrophils in men with obstructive jaundice was found, consisting in an increase in the time to reach the maximum intensity, maximum intensity, area under the curve and activation index for both spontaneous and luminol-dependent induced chemiluminescence. The development of the tumor process in this category of patients was accompanied by a decrease in the area parameter under the curve during spontaneous and induced reactions, time to maximum, intensity maximum and activation index during spontaneous chemiluminescence. The data obtained indicate a marked increase in the values of the functional activity of neutrophils in patients with obstructive jaundice of benign origin, as well as a sharp decrease in their values in the presence of a pathological process of malignant origin.

The functional activity of neutrophils in paranoid schizophrenia and Alzheimer's disease

To evaluate the radical-producing function of neutrophils in paranoid schizophrenia and Alzheimer's disease. The study included 40 patients with paranoid schizophrenia and 22 with Alzheimer's disease. To assess the functional activity of neutrophils, whole blood samples were analyzed using the chemiluminescent method with two-step cell stimulation. Indicators of radical-producing activity of neutrophils in patients were compared to those in age-matched healthy people. The quantitative indicators of radical-producing activity of neutrophils in the paranoid schizophrenia group and in the Alzheimer's disease group correspond to reference intervals of healthy donors, however almost one third of patients with paranoid schizophrenia had an altered shape of the chemiluminescent curve. The neutrophil immune response might be involved in the pathogenesis of paranoid schizophrenia only in some cases.

Оценка изменений концентраций цитокинов и активности нейтрофилов у больных эссенциальной артериальной гипертонией в зависимости от пораженных органов-мишеней на фоне антигипертензивной терапии

Оценка изменений концентраций цитокинов и активности нейтрофилов у больных эссенциальной артериальной гипертонией в зависимости от пораженных органов-мишеней на фоне антигипертензивной терапии

Hazardous effect of the maintenance dose of iron sucrose versus iron dextran on oxidative burst activity of neutrophils in patients on regular hemodialysis; a pilot study

Introduction: Intravenous (IV) iron which is a potent pro-oxidant may decrease the phagocytic function of neutrophils and subsequently leads to repeated bacterial infections in patients on chronic regular hemodialysis (HD).Objectives: We aimed to evaluate the effect of IV iron dextran versus iron sucrose on oxidative burst activity of neutrophils in HD patients.Patients and Methods: A crossover study was included 20 prevalent HD patients were randomly divided into two groups; 10 patients each. Each group received a single dose of 100 mg iron sucrose or iron dextran then shifted to the other type after 4 weeks as a washout period. Patients with evidence of acute infections, diabetes mellitus, chronic liver disease, active collagen disease or any other acquired immune deficiency diseases were excluded. Patients on immunosuppressive drugs or with hypersensitivity to iron therapy or evidence of iron overload (TSAT > 50% and/or ferritin > 800 ng/mL) were also excluded. Erythropoietin maintenance dose was given to all patients. Urea reduction ratio (URR %), C-reactive protein (CRP) titer, complete blood picture and iron study were conducted. Neutrophil oxidative burst test was performed by flow-cytometry with an estimation of stimulation index before and after IV iron dosage. Results: Twenty patients (9 males, 11 females; mean age 49.40 ± 9.02 years) on chronic HD for a mean time of 10 ± 7.32 years. After 100 mg of IV iron infusion in HD patients, a highly significant difference in oxidative burst test of neutrophil cells before and after administration of iron regardless of its type was detected (P < 0.01). This change over time is not significantly different between two types of iron (P > 0.05). However, a significant increase in CRP titer after administration of iron dextran was seen (P = 0.016). Conclusion: Maintenance dosage of either IV iron sucrose or iron dextran (100 mg) had a hazardous effect on the neutrophilic phagocytosis demonstrated by the significant rise in the oxidative burst index in HD patients.

SFasL-mediated induction of neutrophil activation in patients with type 2 diabetes mellitus

Fas/Fas ligand system was shown to be related to insulin resistance and type 2 diabetes mellitus (T2DM). However, the role of soluble Fas ligand (sFasL) in functioning of immune cells in type 2 diabetes mellitus (T2DM) has not been studied yet. The aim of the present study was to determine in vitro effects of sFasL on neutrophil activation and apoptosis. We demonstrate here that sFasL exhibited proinflammatory effect and induced mRNA levels of caspase-1, NF-κB, IL-1β and CD18 expression. At the same time, sFasL induced reactive oxygen species (ROS) production. Activation of caspase-1 activity abolished sFasL-dependent apoptosis, and suppressed Fas expression and mRNA levels of caspase-3 in neutrophils from T2DM patients. Collectively, our findings identify a novel proinflammatory role of sFasL in T2DM neutrophils that is dependent of caspase activity. Thus, sFasL enhances inflammatory response of neutrophils from T2DM patients without increasing apoptosis suggesting its triggering role in T2DM inflammation.

Correlation of Nucleosomes, MMP‐9, and IL‐1α with Relation to Neutrophil Activation in Patients with Atrial Fibrillation

IntroductionAtrial fibrillation (AF) is currently the most common cardiac arrhythmia encountered in clinical practice and a major cause of morbidity and mortality among adults. It is estimated that atrial fibrillation affects approximately 2.7 million people in the United States. Extracellular nucleosomes in plasma (PNs) are complexes of DNA and histones that are released during cell death. Histones have been shown to activate neutrophils by functioning as DAMPs when they are translocated from the nucleus to the extranuclear space. These nucleosomes mediate inflammatory and thrombotic responses and could serve as biomarkers in AF. MMP‐9 degrades type IV collagen and acts as a chemotactic agent for neutrophilic migration. IL‐1α is an acute phase cytokine released by neutrophils that promotes inflammation.ObjectiveTo examine the relationships of circulating levels of PNs, MMP‐9 and IL‐1α in patients with AF.MethodsIn a prospective study, plasma samples were collected from 50 patients prior to ablation surgery for AF at Loyola University Medical Center. ELISAs kits were used as per kit instructions for PNs, MMP‐9 and IL‐1α.ResultsCirculating levels of PNs were positively correlated with levels of MMP‐9 (p=0.006, r=0.49) and IL‐1α (p=0.008, r=0.50) in patients with AF. Additionally, a positive correlation was seen between circulating levels of MMP‐9 and IL‐1α (p=0.02, r=0.41).DiscussionExtracellular nucleosomes function as DAMPs by binding to receptors and triggering the activation of multiple signal pathways. Due to the involvement of inflammation and thrombosis in the pathology of atrial fibrillation, the positive correlation of nucleosome levels with the levels of MMP‐9 and IL‐1α in atrial fibrillation imply PNs may activate the immune system and generate the inflammatory response. These results may provide insight into a novel mechanism of thrombogenesis in patients with AF.

Functional Activity of Neutrophils in Burn Sepsis

A complex effect of thermal trauma on the immune system triggers the tendency to develop and generalize infection in patients with severe thermal trauma. The phagocytic system, which is represented, first of all, by neutrophilic granulocytes, is significantly altered. Objective: to determine the dynamics of changes in the functional activity of neutrophils in patients with thermal trauma, as well as its prognostic significance in the development of purulent-septic complications of a burn disease. Materials and methods. The functional activity of neutrophils was assessed by spontaneous and induced chemiluminescence responses of whole blood sequentially stimulated with phorbol-12-myristate"13-acetate(PMA) and N-formyl-methionyl-leucyl-phenylalanine (fMLP). 34 patients with a burn disease of different severity were dynamically examined during observation. Results. The newly developed technique was applied to study the responses of whole blood of patients with burn disease to stimuli; data on the functional activity of neutrophils depending on various severities of the disease and response dynamics were obtained. Analysis of chemiluminescent curves of the development of respiratory explosion of neutrophils helped to reveal parameters of the immune system state, namely: a one— or two-phase response to a stimulus, a «rapid phase» neutrophil specific activity, and a «slow phase» neutrophil specific activity. Conclusion. The results revealed trends in alterations of functional activity of neutrophils in burn disease. Diagnostic and prognostic indices of the functional activity of neutrophils for assessing the severity of the disease and the prognosis of the septic process were suggested.

Elevated Levels of Circulating Nucleosomes Are Not Associated with Venous Thrombosis or Neutrophil Activation in Patients with Multiple Myeloma

Background: Patients with multiple myeloma (MM) are at a 9-fold increased risk of venous thromboembolism (VTE). Mechanisms underlying the increased VTE risk in MM have not yet been clarified. Recently, extracellular DNA, e.g. nucleosomes, has been shown to play a central role in coagulation activation and propagation in vivo and in animal models. Increased nucleosome levels were demonstrated in patients with VTE, supporting their role in VTE development. The prothrombotic activity of nucleosomes may be of importance in systemic inflammatory states, in which activated immune cells can release nucleosomes, such as neutrophils by forming neutrophil extracellular traps (NETs). However, during inflammation and upon chemotherapy, parenchymal cells and tumor cells, respectively, can also release extracellular DNA in the form of nucleosomes, which could promote coagulation. As MM is regarded as a systemic inflammatory disease, we hypothesized to find elevated levels of nucleosomes in MM patients, with an association between the highest levels of nucleosomes and development of VTE. Therefore, we assessed levels of nucleosomes and neutrophil activation, and their association with VTE, in a cohort of patients with newly diagnosed MM.Methods: We assessed levels of circulating nucleosomes and systemic neutrophil activation, by presence of elastase-α1-antitrypsin (EA) complexes, in 131 patients with newly diagnosed MM using plasma samples. Interleukin (IL)-6 and IL-8 levels were also assessed to estimate the inflammatory state. We used a case-cohort design to investigate the association between nucleosomes, neutrophil activation and VTE, by calculating odds ratios (ORs) with corresponding 95% confidence intervals (CIs) using nucleosomes and EA complexes as categorical variables, based on the 80th percentile for MM patients without VTE, with ≤80th percentile being the reference category. To compare levels of nucleosomes and neutrophil activation complexes in MM patients with patients with other malignancies, a comparison was made with a cohort of cases with objectified deep-vein thrombosis (DVT) of the leg and screened controls without DVT, stratified by malignancy status.Results: 19 of 131 MM patients (14.5%) developed VTE during MM treatment. Levels of nucleosomes (median 39.5 U/mL; interquartile range [IQR] 7.9-156.8, vs. median 25.6 U/mL; IQR 10.9-76.4; p=0.61) and neutrophil activation, as evidenced by presence of EA complexes (median 30.1 U/mL; IQR 22.6-39.3, vs. median 31.6 U/mL; IQR 24.3-46.0; p=0.45), did not differ between MM patients with and without VTE. No association was found between high levels (>80th percentile of non-VTE MM patients) of nucleosomes or neutrophil activation, and VTE (OR 0.5; 95% CI 0.2-1.6, and OR 2.1; 95% CI 0.5-9.7, respectively) in MM patients. IL-6 and IL-8 levels were low in all MM patients, irrespective of VTE. Nucleosome levels were similarly high in MM patients compared with DVT patients with other malignancies, and significantly higher compared with DVT patient without malignancies, and with DVT controls, irrespective of malignancy status (Fig. 1A). In contrast, levels of neutrophil activation were significantly lower in MM patients in all comparisons (Fig. 1B).Conclusions: Elevated levels of nucleosomes were found in MM patients, but without correlating high levels of neutrophil activation. Nucleosome levels in MM patients with and without VTE were similar, and there was no association between high levels of circulating nucleosomes or neutrophil activation, and VTE in MM patients. Our results suggest that elevated levels of nucleosomes in MM patients are independent of neutrophil activation or inflammatory status, but rather a marker of plasma cell injury or death. This could also explain absence of a clear association with VTE. Future studies are necessary to confirm the source and the clinical relevance of high levels of nucleosomes in MM patients. [Display omitted] DisclosuresLeebeek:Baxter: Research Funding; UniQure: Consultancy; Netherlands Hemophilia Foundation: Research Funding; CSL Behring: Research Funding.

Increased Inflammatory Properties of Neutrophils in Patients with Venous Thromboembolism and the Effect of Simvastatin on Abrogating Inflamed Neutrophils Adhesiveness

Neutrophils have a complex migrating process out of the vascular lumen, that consists of chemoattraction and rolling, followed by firm attachment and migration to extravascular tissues. In these sites, neutrophils are capable of promoting phagocytosis, secreting proteases, generating reactive oxygen species (ROS), and probably releasing neutrophil extracellular traps (NETs). Furthermore, neutrophil activation also induces major tissue injury associated with acute and chronic inflammatory disorders, such as the venous thromboembolism (VTE). Recently, animal models and clinical studies in acute VTE have explored the participation of neutrophils in the pathophysiology of VTE. However, VTE has been associated with a chronic inflammatory condition, and it remains unclear whether the activation of neutrophils is persistent after the acute phase of the disease. Furthermore, there are clinical evidences supporting that simvastatin may prevent VTE, since the drug has pleiotropic anti-inflammatory effects. The aim of this study is to evaluate the occurrence of neutrophil activation in patients with VTE compared to health controls and determines the effect of simvastatin in these adhesive properties of the neutrophils. Neutrophils were separated from blood collected over Ficoll-Paque densities. Neutrophils activation was determined by the expression of activated adhesive molecules (LFA-1/CD11a and MAC-1/CD11b) and ROS generation, detected by flow cytometry. Chemotaxis assays (chemoTx, Neuro Probe, Inc) and serum nucleosome, a marker of NETs, were quantified by optical density (OD) (Cell Death Detection ELISAPLUS Kit, Roche). Serum high sensitive CRP (hs-CRP) levels were measured in BN ProSpec System (Siemens) by nephelometry. For CD11a and CD11b integrins expression, cells were evaluated under basal conditions and after TNFα inflammatory stimulus, pretreated, or not, with simvastatin. For the migration assays, neutrophils were treated, or not, with IL-8, a neutrophil chemotactic factor. The results were displayed as mean and standard deviation (±SD). The study group consisted of thirty-seven patients with personal history of VTE, the median time since VTE occurred was 25 months (range 13 - 42 months), the event was spontaneous in 51.35% of the cases and 23 patients presented proximal VTE. Thirty-seven controls, matched with patients according to age, gender and ethnicity, were also included. The mean fluorescence intensity (MFI) of CD11a was higher in VTE patient neutrophils, both in basal conditions (30.84 ± 6.82 vs. 38.72 ± 22.75, P= 0.04) and after TNFα stimulus (34.09 ± 9.64 vs. 45.65 ± 33.06, P= 0.01). Higher MFI of CD11b was observed in patient neutrophils, compared with controls, only after TNFα stimulus (149.10 ± 52.74 vs. 200.0 ± 100.5, P= 0.02), and the stimulus was reverted by pre-treatment with simvastatin (200.8 ± 100.50 vs. 174.60 ± 80.63, P= 0.001). The amount of ROS (MFI) was similar in patients and controls (908.30 ± 423.7 vs. 844.0 ± 312.0, P= 0.83). Neutrophils from VTE patients also presented increased basal chemotaxis (17.55% ± 9.79 vs. 12.64% ± 4.78, P=0.02) and IL-8-stimulated chemotaxis (63.48% ± 29.73 vs. 49.88% ± 19.48%, P=0.06). Serum levels of nucleosomes were similar in patients and controls (1.05 ± 0.81 vs. 0.88 ± 0.62, P= 0.64), however higher levels of circulating nucleosomes were observed in patients with severe post-thrombotic syndrome (PTS), compared to patients with non-severe PTS, without PTS and controls (1.49 ± 0.81 vs. 1.06 ± 0.64 vs. 0.64 ± 0.60 vs. 0.88 ± 0.62, P=0.04). Furthermore, serum levels of hs-CRP were significantly higher in VTE patients when compared with controls (0.59 mg/dl ± 0.58 vs. 0.17 mg/dl ± 0.12, P=0.00). We demonstrated that patients with VTE presented patterns of neutrophil activation long time after the acute thrombotic episode. In particular, the stimuli for neutrophil adhesion and chemotaxis were higher in patients, as detected by the increased activation of adhesive molecules and cell migration. Furthermore, we observed that simvastatin may abrogate the expression of CD11b in inflamed neutrophils. Neutrophil activities associated with ROS generation and the releases of nucleosomes were not increased in these patients. The results may support the hypothesis that increased neutrophils activation is part of the chronic inflammatory condition associated with VTE and may be downregulated by the effects of simvastatin. DisclosuresNo relevant conflicts of interest to declare.

Influence of progesterone on functional activity of neutrophils in threatening preterm labor

Aim of the study. To assess the influence of exogenous progesterone on the functional activity of neutrophils in patients, whose pregnancies proceeded with clinical signs of threatened abortion. Materials and methods. The material used for the study were peripheral blood neutrophils of 32 women, who were hospitalized with a diagnosis of "threatening premature labor" in the term of 22-34 weeks and whose pregnancies later ended in spontaneous preterm (group 1, 10 patients) and term (group 2, 22 patients) birth. Women of 1st and 2nd groups were comparable in age, gestational age and serum concentration of endogenous progesterone. The study was conducted in vitro. Neutrophils were isolated by settling for NBT-test and phagocytosis reaction, and by centrifugation on a double density gradient of ficoll-verografin in order to assess the formation of neutrophil extracellular traps. Neutrophils were added to a suspension of exogenous progesterone at concentrations of 10 ng/ml, 20 ng/ml, 100 ng/ml. The oxidation activity of neutrophils was measured in the spontaneous and stimulated (with Staphylococcus aureus at a concentration of 1×108 bacterial cells in 1 ml) types of NBT-test with microscopic evaluation of the results. The ability of neutrophils to form extracellular traps was evaluated in spontaneous and stimulated (with Staphylococcus aureus at a concentration of 1×108 bacterial cells in 1 ml) types of the test using color dye Hoechst 33342, followed by fluorescent microscopy. Results. Progesterone (10 ng/ml) significantly decreased spontaneous neutrophil oxidative activity [17.0% (12.0-26.0%) and 7.0% (4.0-10.0%), p<0.05] in patients of group 1, but not in patients of group 2. There was no effect of progesterone at studied concentrations on stimulated neutrophil oxidative activity in women of groups 1 and 2. When 20 ng/ml progesterone was added there was a decrease in neutrophil phagocytic index in patients of group 2 [66% (55-77%) and 51% (40-69%), p<0.05]. Progesterone (100 ng/ml) caused an increase in spontaneous NET-forming function in both groups and had no effect on stimulated function. Conclusions. Progesterone is able to provide various immunomodulatory effects on neutrophils of patients, whose pregnancies ended in preterm and term birth. Exogenous progesterone is able to provide various immunomodulatory effects on neutrophils of patients, whose pregnancies ended in preterm and term birth. It reduced the increased spontaneous oxidative activity of neutrophils in women with adverse pregnancy outcomes and lowered phagocytic number in patients with favorable pregnancy outcome. This drug also stimulated spontaneous NET-forming function of the cells, regardless of the pregnancy outcome.

A New Chemiluminescent Method for Evaluation of the Functional Activity of Neutrophils in Patients with Type 2 Diabetes Mellitus.

Functional activity of neutrophils was evaluated by the chemiluminescent method with successive double stimulation by soluble stimuli with different mechanisms of action: phorbol-12-myristate-13-acetate (PMA) and phormyl-methionyl-leucyl-phenilalanine (fMLP). The study was carried out in 26 patients receiving oral sugar-reducing therapy. In addition to the functional activity of neutrophils, the levels of TBA reactive products, inflammation markers, blood clotting values, and biochemical parameters were measured. The results showed mainly reduction of the granulocytic component of the immune system in the patients.

Increased Nucleosomes and Neutrophil Activation Link to Disease Progression in Patients with Scrub Typhus but Not Murine Typhus in Laos.

Cell-mediated immunity is essential in protection against rickettsial illnesses, but the role of neutrophils in these intracellular vasculotropic infections remains unclear. This study analyzed the plasma levels of nucleosomes, FSAP-activation (nucleosome-releasing factor), and neutrophil activation, as evidenced by neutrophil-elastase (ELA) complexes, in sympatric Lao patients with scrub typhus and murine typhus. In acute scrub typhus elevated nucleosome levels correlated with lower GCS scores, raised respiratory rate, jaundice and impaired liver function, whereas neutrophil activation correlated with fibrinolysis and high IL-8 plasma levels, a recently identified predictor of severe disease and mortality. Nucleosome and ELA complex levels were associated with a 4.8-fold and 4-fold increased risk of developing severe scrub typhus, beyond cut off values of 1,040 U/ml for nucleosomes and 275 U/ml for ELA complexes respectively. In murine typhus, nucleosome levels associated with pro-inflammatory cytokines and the duration of illness, while ELA complexes correlated strongly with inflammation markers, jaundice and increased respiratory rates. This study found strong correlations between circulating nucleosomes and neutrophil activation in patients with scrub typhus, but not murine typhus, providing indirect evidence that nucleosomes could originate from neutrophil extracellular trap (NET) degradation. High circulating plasma nucleosomes and ELA complexes represent independent risk factors for developing severe complications in scrub typhus. As nucleosomes and histones exposed on NETs are highly cytotoxic to endothelial cells and are strongly pro-coagulant, neutrophil-derived nucleosomes could contribute to vascular damage, the pro-coagulant state and exacerbation of disease in scrub typhus, thus indicating a detrimental role of neutrophil activation. The data suggest that increased neutrophil activation relates to disease progression and severe complications, and increased plasma levels of nucleosomes and ELA complexes represent independent risk factors for developing severe scrub typhus.

ИЗУЧЕНИЕ ВЛИЯНИЯ НАНОЧАСТИЦ СЕРЕБРА IN VITRO НА УРОВЕНЬ ЭКСПРЕССИИ МОЛЕКУЛЯРНЫХ МАРКЕРОВ АКТИВАЦИИ ЛИМФОЦИТОВ И МАРКЕРА АУТОИММУННОГО ПРОЦЕССА ПЕРИФЕРИЧЕСКОЙ КРОВИ БОЛЬНЫХ ВИРУСНОЙ ПАТОЛОГИЕЙ РОГОВИЦЫ

<p>The influence of the nanoparticles of silver on the expression of molecular markers activation of lymphoid cells CD7<sup>+</sup>, CD25<sup>+</sup>, CD38<sup>+</sup>, CD45<sup>+</sup>, CD54<sup>+</sup>, CD95<sup>+</sup>, CD150<sup>+</sup> and CD5<sup>+</sup> – marker of the autoimmune process, as well as on phagocytic activity of neutrophils in patients with viral pathologies of the cornea was studied <em>in vitro</em>. In the Laboratory of Immunology, SI Institute of Eye Diseases and Tissue Therapy NAMS of Ukraine was developed technique of cultivation of peripheral blood lymphocytes with immunomodulation drugs, followed by determination of changes in the level of expression of molecular markers of lymphocyte activation. Assessment of the level of expression of molecular markers of activation of peripheral blood lymphocytes was performed method using a panel of monoclonal antibodies, CD5<sup>+</sup>, CD7<sup>+</sup>, CD25<sup>+</sup>, CD38<sup>+</sup>, CD45<sup>+</sup>, CD54<sup>+</sup>, CD95<sup>+</sup> and CD 150<sup>+</sup>. The study was conducted <em>in vitro</em> with the peripheral lymphocytes the blood of 23 patients of viral pathology of the cornea. Our studies of the effects of nanosilver particles <em>in vitro</em> on the state of expression of molecular markers of activation of peripheral blood lymphocytes and phagocytic activity of neutrophils in patients with viral corneal pathology, showed a significant increase in the level of expression of the CD7<sup>+</sup>, CD25<sup>+</sup>, CD45<sup>+</sup> and phagocytic activity of neutrophils after application silver nanoparticles.</p> <p><em>Key words: immune system, nanoparticles of silver, monoclonal antibodies, molecular markers activation.</em></p>

Increased metabolic activity of neutrophils in patients with chronic obstructive pulmonary disease.

Aims:To compare the metabolic activity of peripheral neutrophils in patients diagnosed with chronic obstructive pulmonary disease (COPD) with that of healthy, nonsmoking volunteers.Materials and Methods:Venous blood samples were taken from patients diagnosed with COPD as well as from healthy nonsmokers. Each sample was subjected to the nitro blue tetrazolium (NBT) test in which neutrophils exhibiting elevated metabolic activity were detected by light microscopy. The test was repeated after stimulation with Escherichia coli (E. coli) endotoxin with fresh samples. Neutrophils showing dye uptake were then counted in each case.Results:We found that the mean numbers of activated neutrophils without and with the addition of endotoxin were 19% and 23%, respectively, in the control group and 56% and 62%, respectively, in the test group. Two-sample t-test statistic revealed that there was a significant (P < 0.01) increase in neutrophilic metabolic activity in patients with COPD as compared to that in healthy volunteers. This significance remained even after stimulation using E. coli endotoxin.Conclusion:The results hint at a potentially relevant pathogenic mechanism in COPD related to the metabolic activity of neutrophils. By exhibiting enhanced metabolic activity, neutrophils in the COPD patients are more likely to be involved in damaging lung tissues.

In vitro interferon γ improves the oxidative burst activity of neutrophils in patients with chronic granulomatous disease with a subtype of gp91phox deficiency.

Aim of this studyChronic granulomatous disease (CGD) is a genetically heterogeneous primary immunodeficiency caused by a defect in phagocyte production of oxygen metabolites, and resulting in infections produced by catalase-positive microorganisms and fungi. Interferon γ (IFN-γ) has a multitude of effects on the immune system. Although preliminary studies with CGD patients on treatment with IFN-γ showed that it enhanced phagocytosis and superoxide production, ongoing studies did not reveal a significant increase of this function. Here we investigated the oxidative capacity of phagocytes in different subtypes of CGD patients on treatment with IFN-γ in vitro.Material and methodsFifty-seven patients with CGD from 14 immunology centres were enrolled to our multi-centre study. Twenty-one patients were studied as controls. Oxidative burst assay with dihydrorhodamine 123 (DHR) was used and the stimulation index (SI) was calculated with respect to CGD subtypes in both neutrophils and monocytes before, and then one and 24 hours after adding IFN-γ.ResultsUpon comparison of the SIs of the patients’ neutrophils before in vitro IFN-γ at hour 0, and after adding IFN-γ at hour 1 and 24 were compared, and the differences were determined between hours 0-24 and hours 1-24. This difference was especially apparent between hours 1-24. In CGD subtypes, particularly in gp91phox subtype, it was seen that, following in vitro IFN-γ, SIs of neutrophils began to increase after hour 1, and that increase became more apparent at hour 24.ConclusionsOur study showed that IFN-γ treatment may increase the oxidative bursting activity by increasing the superoxide production in neutrophils, particularly in gp91phox subtype.

Functional and metabolic neutrophils activity in patients with acute intestinal infections and the influence of selimakcid

Aim. To explore the functional activity of neutrophils in healthy individuals and patients with acute intestinal infections and the effect of selimakcid on it in vitro. Methods. 48 patients with acute intestinal infections aged 20 to 60 years were examined. The control group consisted of 10 healthy people of the same age. Functional and metabolic activity of neutrophils was studied by a nitro blue tetrazolium chloride test and by «BURSTTEST» (Neutrophil Oxidative Burst Test). Results. Examination of the functional and metabolic activity of neutrophils in patients with acute intestinal infections revealed increased results of spontaneous nitro blue tetrazolium chloride test in acute period of the disease. The same changes were discovered while studying the Serratia marcescens stimulated cell activity in patients with dysentery. However, in patients with salmonellosis neutrophils activity in induced nitro blue tetrazolium chloride test did not differ from that of healthy controls. The number of neutrophils synthesizing reactive oxidants recorded by «BURSTTEST» in patients corresponded to the indicators of the induced nitro blue tetrazolium chloride test and did not differ in patients with different nosologies. Studying the effect of selimakcid on neutrophils functional activity in healthy controls and patients with acute intestinal infections by nitro blue tetrazolium chloride test and «BURSTTEST» showed that pre-incubation of neutrophils with selimakcid intensified the response to Serratia marcescens. The ability of the drug to modulate the neutrophils activity in patients with acute intestinal infections was higher compared to healthy individuals. Conclusion. Increased functional activity of neutrophils associated with reduced reserve capacity was revealed in patients with salmonellosis and shigellosis of moderate severity. Selimakcid showed immunomodulatory effect, increasing the functional and metabolic potential of neutrophils in patients with acute intestinal infections in vitro.

Level of mannose-binding lectin and phagocytic activity of leukocytes in patients with acute coronary syndrome

Mannose-binding lectin (MBL) is a key component of innate immunity that starts one of the ways of complement activation. Factors of neutrophil activation are cell factors of innate and acquired immunity. to study MBL levels and factors of neutrophil activation in patients with acute coronary syndrome (ACS). A total of 45 patients with ST elevation (STE) ACS and non ST-elevation (NSTE) ACS were enrolled in the study, 15 persons were age-matched controls. RESULTS. Compared with control group MBL level was higher in patients with ACS (52.7 vs 127.2 hg/ml, respectively, p = 0.07), and significantly higher in patients with NSTE ACS (52.7 vs. 164.7 hg/ml, p = 0.03). There was no difference between MBL levels in STE ACS and NSTE ACS patients. Patients with inferior myocardial infarction (MI) had significantly higher MBL level than those with anterior MI (182.8 -92.7 hg/ml, p = 0.02). Patients with concomitant diabetes had statistically higher MBL level than patients without diabetes (225 vs 100 hg/ml, OR 2.25, p = 0.03). MBL level was lower in patients with low (<40%) ejection fraction - 92.7 vs 148.9 hg/ml in patients with EF > or = 40% (p = 0.19). No difference of neutrophil activation factors between ACS patients and controls was found (phagocytic activity of neutrophils 74.5 vs 74.3%, phagocytic number 3.34 vs 4.36, phagocytic reserve 88 vs 85.5 in ACS and control group, respectively). Elevated innate immunity factor (MBL) level was associated with ACS, especially in patients with diabetes mellitus. No association between cell immunity factors with ACS was found.