Activity Of Neutrophil Granulocytes Research Articles

Vaccination Against Poly-N-Acetylglucosamine Decreases Neutrophil Activation and Gvhd While Maintaining Microbial Diversity

Allogeneic hematopoietic cell transplantation (allo-HCT) is a curative therapy for different malignant and non-malignant hematologic diseases. Major life-threatening complications after allo-HCT are acute graft-versus-host disease (aGVHD) and severe infections. The incidence of GVHD after allo-HCT remains high, despite prophylactic immunosuppressive medication. According to the CIBMTR database, 60% of patients undergoing allo-HCT develop at least grade II-IV aGVHD. Microbial invasion into the intestinal mucosa after allo-HCT triggers the activation of neutrophil granulocytes (neutrophils) and requires antibiotic treatment to prevent sepsis. However, antibiotics lead to a loss of microbial diversity, which is connected to a higher incidence of aGVHD. Anti-microbial therapies which eliminate invading bacteria and diminish neutrophil-mediated damage without reducing the diversity of the microbiota are therefore highly desirable. A potential solution for this could be the use of anti-microbial antibodies that target and mark invading pathogens resulting in their elimination by innate immune cells. Therefore we investigated the potency of both active and passive immunization against the conserved microbial surface polysaccharide Poly-N-acetylglucosamine (PNAG) which is expressed on various pathogens in a murine aGVHD model. We could detect reduced aGVHD related mortality in mice which underwent treatment with an antibody against PNAG (anti-PNAG) or were actively immunized against PNAG. Sequencing analysis of the microbiota before and after allo-HCT showed that anti-PNAG treatment did not have any effect luminal microbial diversity. Mechanistically, we could show that anti-PNAG antibody treatment caused more abundant neutrophil recruitment to the intestinal submucosa. This supports the concept that neutrophils are able to effectively eliminate any opsonized bacteria in this tissue, leading to reduced local inflammation. In agreement with this we observed lower intestinal inflammation and reduced myeloperoxidase activity as well as proliferation of neutrophils in the small intestine of mice treated with anti-PNAG antibody. We demonstrate the potency of targeting PNAG as a novel antimicrobial treatment option in aGVHD by reducing uncontrolled neutrophil activation and thereby interfering with aGVHD without affecting commensal intestinal microbial diversity. Anti-PNAG-antibodies enhance anti-microbial immunity, while reducing the activation of neutrophil-mediated downstream immunopathology which is a novel approach to reduce the severity of GVHD. This could be translated into a clinical application given the modest toxicity profile of vaccination and the availability of fully human anti-PNAG antibodies, both of which have been tested in phase 1 trials in humans (ClinicalTrials.gov Identifier: NCT02853617). Disclosures Cywes-Bentley: OneBiopharma, Inc.: Honoraria. Koenecke:Novartis: Other: none. Pier:OneBiopharma, Inc.: Equity Ownership, Honoraria; Alopexx Vaccine, LLC: Equity Ownership, Honoraria.

Biological activity of a polyphenolic complex of Arctic brown algae

One of the most significant groups of compounds that determine the pharmacological significance of Arctic brown algae is the polyphenols, namely phlorotannins. Brown algae of the family Fucaceae, growing in the waters of the Arctic seas, accumulate a large number of polyphenolic compounds. The pharmacological significance of plant polyphenols is related to their structure; however, it is worth noting that the relationship between the molecular weight and the biological activity of brown algae phlorotannins is still not well understood. Thus, the aim of this work was the isolation and characterization of the polyphenol fraction of the Arctic brown algae Fucus vesiculosus with the following evaluation of the radical scavenging, bacteriostatic, fungistatic, and phagocytic activities. The highest radical scavenging activity (700–900 mg of ascorbic acid g−1 of extract) has fractions whose composition is characterized by the predominance of oligomeric phlorotannins containing from 3 to 8 structural units of phloroglucinol in the range of 374–994 Da. The polyphenol fractions obtained exhibit bacteriostatic effects in 45–85% of the studied pathogenic bacterial cultures. The fungistatic effect reaches a maximum of 35%. The test samples do not possess immunosuppressive properties and do not inhibit the phagocytic activity of neutrophilic granulocytes of human blood.

Assessment of the functional activity of neutrophilic granulocytes in patients with metabolic syndrome

Assessment of the functional activity of neutrophilic granulocytes in patients with metabolic syndrome

Changes in cellular factors of local immunity udder of cows with mastitis

The immunological aspects of lactation of cows are investigated in this work. The authors present modern scientific data on the local immune protection of the mammary gland of cows. The purpose of the research was to improve the laboratory methods of diagnosis of mastitis and to study the cytological composition of the secretion of the breast of the cows for subclinical and purulent catarrhal inflammation of the udder. The cytological differentiation of milk cells was carried out according to its own developed, tested and patented method. Our experimental data convincingly confirm that in subclinical and purulent-catarrhal mastitis in cows there are significant changes in changes in the cytological composition and the system of local immune defense of the mammary gland. An important role in inflammation of the mammary gland is played by neutrophilic granulocytes that migrate to the parenchyma at the beginning of the clinical pathology (with subclinical inflammation) and release a range of inflammatory mediators in the inflammation zone. Subclinical mastitis of cows is accompanied by a sharp increase in the total somatic cells count in the secretion of the mammary gland. This is due to the activity of neutrophilic granulocytes, monocytes of histiocytes. Corresponding changes were found in the cytological balance of immunocompetent cells of the secretion of cows at from subclinical mastitis. The most significant shift was noted in the index of migratory activity of neutrophils (P < 0.01), which is a specific “indicator” indicator of inflammatory response in the body of diseased animals. Subclinical inflammation was also manifested by a decrease in the migration activity of lymphocytes. At purulent-catarrhal mastitis of cows there is a significant increase in the number of somatic cells. Also, the migration activity of neutrophils and histiocytes in the pathologic process zone is increasing and changes in the balance of immune cells are established. Thus, our experimental data convincingly confirm that in subclinical and purulent-catarrhal mastitis in cows there are significant changes in the system of local immune defense of the mammary gland. It has an important diagnostic value and also needs immunocorrection in the treatment process.

Functional activity of neutrophils and local manifestations of oxidative stress in the mucous membrane of the nasal cavity in the early postoperative period

The purpose of the study was to study the factors of antimicrobial, antioxidant defense, variability of nasal secretion lipid peroxidation in patients with curvature of the nasal septum in the early postoperative period. METHODS: microbiological, immunological (assessment of the activity and intensity of phagocytosis, nitrous tetrazolium reduction test) biochemical (study of content diene conjugates, ketodienes, conjugated trines and Schiff bases in the lipid extract of neutrophilic granulocytes), statistical. RESULTS: The dynamics of the content, functional and biochemical activity of neutrophilic granulocytes in nasal washings in individuals who underwent rhinosurgical intervention testifies to the presence of qualitative and quantitative changes in neutrophilic granulocytes of the nasal secretion, recorded in reactions of oxygen-dependent metabolism, free radical oxidation and antioxidant protection.

Changes in Functional Activity of Neutrophils and Monocytes Isolated from the Peripheral Blood of Women at Different Phases of the Menstrual Cycle.

We studied functional activity of neutrophilic granulocytes and monocytes isolated from the peripheral blood of women during the follicular and luteal phases of the menstrual cycle. It was shown that phagocytic activity of neutrophilic granulocytes increases, their intracellular oxygen-dependent bactericidal activity decreases, and the number of monocyte extracellular traps increases in women in the luteal phase of the menstrual cycle in comparison with the follicular phase.

Peculiarities of the exposure of the antibiotics of the macrolid group on the functional activity of neutrophilic granulocytes of peripheral blood in vitro

Dose-dependent eff ects of antibiotics and their eff ect on immune cells of the body, in particular neu-trophilic granulocytes, are still poorly understood. The method of fl ow cytometry was determined by the minimum eff ective dose of chemomycin by assessing the degranulation of neutrophils. It was found that under the action of minimum eff ective dose chemomycin neutrophils form neutrophil extracellular traps (NETs), which are one of the mechanisms of adaptive immunity. It is noteworthy that the release of NETs in response to the action of chemomycin was not dependent on gender.

Influence of natural and technogenic low-intensity electromagnetic radiation on the phagocytarial activity of neutrophilic granulocytes, their ability to formation of neutrophilic extraculative traces and biological film formation of microorganisms

Modeled NEMI, similar to natural, without aff ecting the persistence of resident representatives of microfl ora, increasing the phagocytic capacity of neutrophilic granulocytes and stimulating the formation of NVL, can participate in enhancing the anti-infection protection of the human body. Otherwise, the technogenic NEMI acts, in which the phagocytic capacity of neutrophils is suppressed, and every 5 of them forms an extracellular network of DNA strands. Technogenic NEMI inhibits the ability to synthesize biofi lm representatives of the normal fl ora of the human body, thereby violating the colonization resistance of the mucous membranes. Technogenic NEMI can contribute to disruption of homeostasis and development of pathological processes in the human body.

Methods of correction of the functional infringements of phagocytes and local manifestations of oxidative stress in the multiple shell of the nose region with use of ultrasound cavitation

The results of studying the eff ectiveness of the use of ultrasound cavitation therapy in patients with curvature of the nasal septum in the early postoperative period are presented. Research methods: microbiological, immunological (assessment of phagocytosis activity and intensity, nitrous tetrazolium reduction test), biochemical (investigation of lipoperoxidation intensity in terms of primary, secondary and fi nal products of lipid peroxidation: diene conjugates, ketodienes and conjugated trines, Schiff bases in lipid extract), statistical. Positive dynamics of functional and biochemical activity of neutrophilic granulocytes in nasal washings in persons who underwent rhinosurgical intervention after ultrasound cavitation therapy is shown, which indicates the presence of qualitative and quantitative changes in neutrophilic granulocytes of nasal secretion, recorded in reactions of oxygen-dependent metabolism, free radical oxidation and antioxidant protection using the method ultrasound cavitation therapy in early postoperative period.

Phasor-Based Endogenous NAD(P)H Fluorescence Lifetime Imaging Unravels Specific Enzymatic Activity of Neutrophil Granulocytes Preceding NETosis.

Time-correlated single-photon counting combined with multi-photon laser scanning microscopy has proven to be a versatile tool to perform fluorescence lifetime imaging in biological samples and, thus, shed light on cellular functions, both in vitro and in vivo. Here, by means of phasor-analyzed endogenous NAD(P)H (nicotinamide adenine dinucleotide (phosphate)) fluorescence lifetime imaging, we visualize the shift in the cellular metabolism of healthy human neutrophil granulocytes during phagocytosis of Staphylococcus aureus pHrodo™ beads. We correlate this with the process of NETosis, i.e., trapping of pathogens by DNA networks. Hence, we are able to directly show the dynamics of NADPH oxidase activation and its requirement in triggering NETosis in contrast to other pathways of cell death and to decipher the dedicated spatio-temporal sequence between NADPH oxidase activation, nuclear membrane disintegration and DNA network formation. The endogenous FLIM approach presented here uniquely meets the increasing need in the field of immunology to monitor cellular metabolism as a basic mechanism of cellular and tissue functions.

Immunological NBT Test – Indicator of Phagocytic and Metabolic Activity of Neutrophilic Granulocytes in Complications of Ent Organs Diseases

Infectious diseases associated with diseases of ENT organs are quite common. Therefore, this problem requires detailed study. The main goal of the work is to characterize the immunological NBT test – the index of phagocytic and metabolic activity of neutrophilic granulocytes in cases of complications of ENT diseases. To achieve this goal and study the problem in detail, the authors used experimental methods. The experiment was conducted on 40 patients from 32 to 65 years. The authors found that during the eruptive phase of the disease, the percentage of NBT positive granulocytes and tetrazolium activity significantly increased and amounted to 30-40%, while the control group’s data were the following – the activity of the NBT test was 4.9-6.8%. It was also found that the NBT test in patients with otitic meningitis was 26-62%, while in all patients with serous meningitis, the level of the NBT test was low – 6-11.

ВПЛИВ МАЗІ, ЩО МІСТИТЬ СУХИЙ ЕКСТРАКТ КОРЕНЯ СОЛОДКИ, НА ФАГОЦИТАРНУ АКТИВНІСТЬ ГРАНУЛОЦИТАРНИХ НЕЙТРОФІЛІВ КРОВІ В МУРЧАКІВ З ЕКСПЕРИМЕНТАЛЬНИМ ДЕРМАТИТОМ

У статті наведено результати вивчення впливу мазі «Алерголік», що містить сухий екстракт кореня солодки, на фагоцитарну активність гранулоцитарних нейтрофілів крові мурчаків з експериментальним алергічним контактним дерматитом, викликаним 2,4-динітрохлорбензолом. Згідно з отриманими даними, мазь «Алерголік», що застосовувалась в умовах лікувально-профілактичного режиму, мала виражену імунотропну дію, яка полягала в достовірному зниженні, відносно групи контрольної патології, фагоцитарної активності гранулоцитарних нейтрофілів, чого не спостерігали в групі тварин, яких лікували препаратом порівняння «Фладекс».

FEATURES OF FREE RADICAL OXIDATION AND ANTIOXIDANT PROTECTION IN CHILDREN WITH SUGAR DIABETES OF THE FIRST TYPE

Aim. Assessment of the state of lipid peroxidation, antioxidant defense and oxidative stress indicators on the model of functional activity of neutrophil granulocytes of blood in children with type 1 diabetes mellitus depending on the stage of endocrinopathy compensation. Materials and methods. The material of laboratory-diagnostic and clinical studies was the results of a survey of 38 children without endocrine pathology and 89 children with type 1 diabetes mellitus at various stages of disease compensation. The state of antioxidant protection was assessed taking into account the overall antioxidant activity, superoxide dismutase activity, retinol level, α-tocopherol, the content of reduced and oxidized glutathione. The intensity of lipoperoxidation processes was studied by the level of substrates with conjugated double bonds, diene conjugates, ketodienes, conjugated trienes, malonic dialdehyde, common lipids. Functional activity of neutrophil granulocytes was investigated by means of luminol-dependent chemiluminescence. Results. In children with type 1 diabetes mellitus, the intensification of lipid peroxidation processes and the activation of the antioxidant defense system, including variously directed changes in non-enzymatic mechanisms, are noted against the negative effect of active oxygen species. In children with decompensated type 1 diabetes mellitus, severe lipid metabolism disorders combined with oxidative stress indices increase the course of endocrine pathology, significantly increasing the likelihood of intravascular complications. Conclusion. The coordinated generation of active forms of oxygen and the processes of oxygen-dependent metabolism of blood cells in children with compensated type 1 diabetes mellitus indicates the development of the second stage (resistance) of oxidative stress. Reduction of reactive oxygen species production, reduced activation rate оxygendependent metabolism phagocytes incompleteness phagocytosis mechanisms, correlating with an increase in lesion area (destruction of insulin-producing β-cells) of the pancreas in children with decompensated type 1 diabetes, indicating the occurrence of the third step (debilitation) oxidative stress. Violations of metabolic pathways in children with type 1 diabetes are determined by the intensity of «respiratory burst» of neutrophil granulocytes in the «Lipid peroxidation – antioxidant protection».

The effect of continuous low-intensity laser irradiation of the red spectrum on the changes in the functional activity and speed of NADPH-oxidase response of human peripheral blood neutrophils

Registration of the changes in the functional metabolic status of neutrophils under the influence of low-intensity laser radiation (LILR) can be useful for the choice of the exposure parameters in the studies designed to evaluate immunotropic effects. Such investigations, in their turn, are very promising in clinical terms since their results can be used to optimize the laser therapy techniques. The objective of the present study was to evaluate the degranulation potential of neutrophils and the intensity of their NADPH-oxidase reaction in response to the exposure to laser radiation as exemplified by the release of lysosomal granules in vitro. The source of radiation was the LASMIK device operated in the continuous mode at the following parameters: wavelength 635 nm, power density 0.12 mW/cm2, exposure time 10, 30, 90, 120, 150 s in the case of the automatic timer control and 100 s in the case of manual shutdown. It was shown that the maximum lysosomal activity and release of lysosomal granules took place at a 90-10 s exposure and a wavelength of 635 nm with the appearance of a plateau within 120 and 150 s after the onset of irradiation. In the case of a shorter exposure of neutrophil granulocytes to LILR (10 and 30 s) no pronounced effect was observed. It means that low-intensity laser radiation of the red spectrum with a wavelength of 635 nm is a physical stimulus reinforcing exocytosis of lysosomal granules by neutrophils in vitro. The reliable changes of NADPH-oxidase activity of neutrophil granulocytes isolated from donor peripheral blood are recorded at an optimum exposure time of 90-100 s. It is concluded that the laser therapy techniques, at least those designed to regulate neutrophils should be applied with the optimum exposure time of 90-100 s.

Pathophysiology of peritonitis

Despite intensive research efforts peritonitis leading to subsequent sepsis remains associated with a high mortality. The initial effector cells are the locally residing cells of the peritoneum, such as mesothelial cells, mast cells, macrophages and lymphocytes. Through the secretion of chemokines, an influx of neutrophils initially takes place followed by monocytes. The latter can differentiate into inflammatory macrophages. The non-directed activity of neutrophilic granulocytes is limited by the induction of apoptotic programs. Through the breaching of cytokines, bacteria and microbial products into the circulation, a systemic reaction in the sense of systemic inflammatory response syndrome (SIRS) or sepsis arises. This is viewed as a concomitant derailing of inflammatory as well as anti-inflammatory responses, which leads to extensive apoptosis of lymphocytes. The presentation of apoptotic cells leads to a strong immunosuppression. Due to the coexistence of hyperinflammation and immunosuppression, exact knowledge of the current immune status of the patient is a prerequisite in the development of immunotherapies for the treatment of sepsis.

HEMOSTASIS AND FUNCTIONAL PROPERTIES OF NEUTROPHILS IN PATIENTS WITH VARIOUS SENSITIVITY TO ACETYLSALICYLIC ACID IN ACUTE CORONARY SYNDROME

Aim. To study the specifics of relation between hemostasis parameters and functional activity of neutrophil granulocytes in sensitive and resistant to acetylsalicylic acid (ASA) patients with acute coronary syndrome. Material and methods. Totally 53 patients included during the first 24 hours of ACS. Controls consisted of 50 relatively healthy individuals. All patients before the treatment and revascularization were assessed for the resistance to ASA, and selected into groups of sensitive (AS) and resistant (AR). Sensitivity test was done in vitro. In all participants the coagulation and platelet clotting systems were studied. With the luminescent method the neutrophil activity was assessed. Results. In AR with ACS there was decrease of platelets in blood, decreased level of ADP- (ADP 5 mcM) and adrenalin-induced aggregation, increases the level of ADP-induced platelets aggregation (PA) with ADP 0,1 mcM, and von Willebrand factor level. In the ASA sensitive patients there was a decrease of adrenalin induced aggregation. There were no specifics of coagulation hemostasis according to ASA sensitivity. Chemiluminiscence (CL) of neutrophils in ACS has a maximized out time. In AR there was decrease of activation. In AS functional activity of neutrophils more significantly correlates with the parameters of clotting, but in AR — with fibrinolytic and anticoagulation parameters. Conclusion. In ACS patients depending on ASA sensitivity, there are character specifics of platelet clotting: increase of ADP-induced aggregation in AR, decrease of platelet amount and the level of von Willebrand factor. In ACS patients there is a decreased velocity of respiratory boost in neutrophils. In AR there is more prominent dysbalance of coagulation hemostasis and respiratory boost of neutrophils, with the decrease of neutrophil reactivity related to clotting activity and appearance of relations with the parameters of fibrinolytic and anticoagulation activity.

Stress-induced adaptation of neutrophilic granulocyte activity in K and R3 carp lines

Both in mammals and fish, stress induces remarkable changes in the immune response. We focused on stress-induced changes in the activity of neutrophilic granulocytes in the R3 and K lines of common carp, which showed differential stress responses. Our study clearly demonstrates that a prolonged restraint stress differentially affects the activity of K and R3 carp neutrophils. In the K line, stress decreased the respiratory burst, while in the R3 line it reduced the release of extracellular DNA. Surprisingly, the stress-induced changes in ROS production and NET formation did not correlate with changes in gene expression of the inflammatory mediators and GR receptors. In neutrophilic granulocytes from K carp, gene expression of the stress-sensitive cortisol GR1 receptor was significantly higher than in neutrophils from R3 fish, which will make these cells more sensitive to high levels of cortisol. Moreover, upon stress, neutrophilic granulocytes of K carp up-regulated gene expression of the anti-inflammatory cytokine IL-10 while this was not observed in neutrophilic granulocytes of R3 carp. Therefore, we can hypothesize that, in contrast to R3 neutrophils, the more cortisol sensitive neutrophils from K carp respond to stress with up-regulation of IL-10 and consequently reduction of ROS production. Most probably the ROS-independent NET formation in K carp is not regulated by this anti-inflammatory cytokine. These data may indicate a predominantly ROS-independent formation of NETs by carp neutrophilic granulocytes. Moreover, they underline the important role of IL-10 in stress-induced immunoregulation.

Correction: Mesenchymal Stem Cells Augment the Anti-Bacterial Activity of Neutrophil Granulocytes

Correction: Mesenchymal Stem Cells Augment the Anti-Bacterial Activity of Neutrophil Granulocytes

Mesenchymal stem cells augment the anti-bacterial activity of neutrophil granulocytes.

BackgroundMesenchymal stem cells (MSCs) participate in the regulation of inflammation and innate immunity, for example by responding to pathogen-derived signals and by regulating the function of innate immune cells. MSCs from the bone-marrow and peripheral tissues share common basic cell-biological functions. However, it is unknown whether these MSCs exhibit different responses to microbial challenge and whether this response subsequently modulates the regulation of inflammatory cells by MSCs.Methodology/Principal FindingsWe isolated MSCs from human bone-marrow (bmMSCs) and human salivary gland (pgMSCs). Expression levels of TLR4 and LPS-responsive molecules were determined by flow cytometry and quantitative PCR. Cytokine release was determined by ELISA. The effect of supernatants from unstimulated and LPS-stimulated MSCs on recruitment, cytokine secretion, bacterial clearance and oxidative burst of polymorphonuclear neutrophil granulocytes (PMN) was tested in vitro. Despite minor quantitative differences, bmMSCs and pgMSCs showed a similar cell biological response to bacterial endotoxin. Both types of MSCs augmented anti-microbial functions of PMNs LPS stimulation, particularly of bmMSCs, further augmented MSC-mediated activation of PMN.Conclusions/SignificanceThis study suggests that MSCs may contribute to the resolution of infection and inflammation by promoting the anti-microbial activity of PMNs. This property is exerted by MSCs derived from both the bone-marrow and peripheral glandular tissue.

IMMUNOLOGICAL PARAMETERS IN PATIENTS WITH MONOINFECTION BY HUMAN PAPILLOMAVIRUS AND IN PATIENTS CO-INFECTED BY PAPILLOMAVIRUS AND UROGENITAL PATHOGENS

Aim of this study was to investigate the characteristics of cellular and humoral immunity and functional activity of neutrophilic granulocytes in women with monoinfection by human papillomavirus and combination of human papillomavirus (HPVI) and urogenital pathogens (UGI). It was established that more serious disorders in the immune system in women were revealed in case of mixed HPVI and UGI in compare with case of monoinfection by human papillomavirus. Only in patients with combined HPVI and UGI the content of total lymphocytes, T-cells and cytotoxic T lymphocytes was reduced. In patients with combined HPVI and UGI the humoral immunity was characterised by increased concentration of IgM, its level of relative synthesis and content of circulating immune complexes. Independently of the infection type in women the reduction of CD4+ lymphocytes, NK-cells, increased number of γδТ-lymphocytes, as well as increase the functional activity of blood neutrophils have been detected. However, in case of combined HPVI and UGI an inhibitory effect on the reserve capacities of phagocytic cells was revealed.