Activity Of Neutrophil Granulocytes Research Articles

Disruption of the Functional Activity of Neutrophil Granulocytes as a Risk Factor for the Development of Lung Damage in Pregnant Women with COVID-19.

Currently, the assessment of immune status in patients with COVID-19 is limited to determining the count of polymorphonuclear leukocytes and the phagocytic function of neutrophils, which is insufficient to understand the regulatory role of innate immunity cells in the development of pneumonia. However, no such studies have been conducted in pregnant women with COVID-19. The aim of this study was to investigate the functional state of neutrophil granulocytes in order to identify predictors of pneumonia severity risk in pregnant women with COVID-19. A clinical characterization of pregnant women with COVID-19 in addition to minimal and average lung changes was provided. The composition and ratio of morphological forms of leukocyte cells were studied. Cytochemical studies of neutrophil granulocytes were carried out and calculations of the mean cytological index (MCI) for succinate dehydrogenase, myeloperoxidase, and cationic proteins were performed. The number of NETs in blood smears was counted. Independent predictors of pneumonia severity in pregnant women with COVID-19 were calculated using regression analysis. The quality of the model was assessed using ROC analysis. In pregnant women with COVID-19 and an average volume of lung changes, the number of polymorphonuclear leukocytes (p = 0.03) and band neutrophils (p = 0.002) in the blood was significantly higher than in pregnant women with minimal lung changes. The MCI indicators of succinate dehydrogenase, cationic proteins, and myeloperoxidase in pregnant women with COVID-19 were reduced in relation to the control group (p < 0.0001). In blood smears of pregnant women with COVID-19 and an average volume of lung changes, the number of NETs increased (p = 0.002). Regression analysis showed that succinate dehydrogenase and NETs are independent predictors of pneumonia severity in pregnant women with COVID-19. Our study confirms the prognostic significance of low levels of neutrophilic succinate dehydrogenase and high levels of NETs in the blood of pregnant women with COVID-19. The combination of these two biomarkers is a significant reflection of the severity of pneumonia development in pregnant women with COVID-19. However, further research is needed to identify the mechanisms underlying this association.

Prediction of the risk of recurrence of the leprosy process

The aim of the study was to study the degree of neutrophil granulocyte myeloperoxidase activity in peripheral blood of patients with leprosy in order to predict the possible recurrence of the infectious process.Materials and methods. Electron grams of biopsies of the affected skin and blood smears of 8 (5 women and 3 men) patients with borderline (BB), 9 (6 women and 3 men) patients with subpolar lepromatous (LLs) and 7 (4 women and 3 men) patients with polar lepromatous (LLr) were used as the object of the study forms of leprosy. At the ultramicroscopic level, the degree of MP activity detected by the electron-cytochemical method in the Mf of biopsies of the affected skin, both first registered before the start of therapy and with the activation of the process during treatment, was taken into account. The percentage of mitochondria with MP Mf activity relative to the MP of inactive cells was calculated. In peripheral blood smears, the activity of neutrophilic granulocytes (NG) MP was evaluated by the mean cytochemical coefficient (SCC).Results. It was found that in patients with the highest percentage of peroxidase-active mitochondria (79-92%), the presence of MP activity on the membranes of phagosomes and in the electron-transparent zone (around M. leprae) correlated with rapid and persistent regression of the leprosy process and long periods of relapse-free period, which was determined by comparing the data of medical histories with the results obtained in during our research. It was found that in patients without relapse with forms of BB and LLs leprosy, the activity of MP in neutrophils was 1.3 times higher (p&lt;0.05), and the percentage of peroxidase-active mitochondria was 3 times higher (p&lt;0.01) compared with these indicators of patients with relapse of the disease. When comparing the studied markers of groups of patients without relapse with the form of leprosy LLp, the degree of MP NG activity increased by 1.4 times (p&lt;0.05), the percentage of mitochondria with MP Mf activity increased by 3 times (p&lt;0.01) relative to patients with relapses.Conclusion. Thus, the determination of myeloperoxidase activity of neutrophil granulocytes of peripheral blood can act as a prognostic marker for determining the effectiveness of anti-leprosy therapy and the risk of relapses of the disease during and after treatment.

Number of blood neutrophil granulocytes is associated with diffuse coronary involvement in patients with increased lipoprotein (a)

Aim. To assess the level of lipoprotein (a) (Lp(a)) and other lipid profile parameters, the composition of blood leukocytes in patients with focal and diffuse coronary atherosclerosis who underwent coronary artery bypass surgery (CABG), as well as to study the effect of Lp(a) on the activation of leukocytes in vitro.Material and methods. The study included 120 male patients (mean age, 67±5 years) with multivessel coronary artery disease after elective CABG surgery. According to the type of coronary artery involvement, patients were divided into groups of diffuse (n=77) and focal (n=43) atherosclerosis. In peripheral blood samples, the content of the main fractions of lipoproteins, including Lp(a), was determined. Activation of neutrophil granulocytes upon addition of Lp(a) was assessed in whole blood and in primary cell culture.Results. In patients with diffuse coronary stenosis, there was a tendency to higher Lp(a) levels; no changes in other lipid profile parameters were detected. In this group of patients, an increase in the absolute leukocyte count was noted due to neutrophilic granulocytes. An absolute neutrophil count &gt;2,9 million/mL was associated with diffuse disease (AUC, 0,69 (95% confidence interval (CI) 0,600,77), p=0,0002, sensitivity 85%, specificity 42%). In patients with Lp(a) ³30 mg/dL, the diagnostic value of a neutrophil count &gt;2,9 million/mL for diffuse stenosis was higher (AUC, 0,74 (95% CI 0,60-0,86), p= 0,005, sensitivity 90%, specificity 48%). According to logistic regression analysis with the introduction of neutrophils and conventional risk factors (age, body mass index, hypertension and smoking) into the model, only the number of neutrophils above the threshold level remained an independent factor in diffuse coronary stenosis. Lp(a) can induce neutrophil activation, detected by increased CD66b overexpression in whole blood, as well as the formation of neutrophil extracellular traps in primary cell culture.Conclusion. Neutrophil granulocytes can participate in the formation of diffuse atherosclerotic coronary lesions. One of the neutrophil activators may be Lp(a). The combination of high neutrophil count and Lp(a) level in the blood increases the risk of diffuse coronary atherosclerosis.

Immune system dysfunction in purulent inflammatory diseases of the maxillofacial area in pediatric patients

The issues of treatment of purulent inflammatory diseases of the maxillofacial region (PMMA) are quite urgent, due to increasing number of such patients. Their clinical course is getting worse, and the efficiency of antibiotic therapy is decreasing. Clinical outcomes of purulent maxillofacial pathology in children are complicated by potential severe local deformities at the growth areas of the jaw bones which are difficult to eliminate. The number of cases of prolonged and chronic inflammatory processes, development of local and general complications is increased. The reason for these complications may be an impaired susceptibility to infectious agents, which is determined by the state of the immune system. Therefore, our aim was to reveal some features of immune functions in children with purulent-inflammatory diseases of maxillofacial area.&#x0D; The study included a group of pediatric patients 8-17 years old with maxillofacial inflammatory diseases of the maxilla (the study group), and 13 conditionally healthy children (the comparison group). The contents of T cells (CD3+CD19-, CD3+CD4+, CD3+CD8+, CD3+CD4+/CD3+CD8+), and В cells (CD3-CD19+), NK (CD3-CD16+CD56+) was determined using Cytomics FC-500 (Beckman Coulter, USA);concentrations of serum IgA, IgM, IgG were determined by ELISA technique (Vector-Best, Russia). Phagocytic activity of neutrophilic granulocytes (NG) was evaluated as percentage of actively phagocytic NGs, capturing processes were assessed by appropriate phagocytic indices, and the digestive activity was evaluated against S. aureus (strain 209).&#x0D; Combined defects of immune response in children with maxillofacial hypertension were established: decrease of T lymphocytes contents along with decrease of T helpers and CTL ratio along with unchanged content of NK cells and B lymphocytes. Increase of IgA and IgG levels was also found. Defects of phagocytosis were revealed, primarily connected with the processes of completed phagocytosis and increased content of actively phagocytizing NG.&#x0D; Treatment of children suffering with purulent inflammatory diseases of maxillofacial region is still an urgent problem in dentistry. The revealed dysfunction of immune response to pathogens in the purulent maxillofacial disorders may explain a prolonged clinical course of inflammatory processes, thus determining a need for usage of immunotropic therapy in complex treatment schedules including operative aid as well as conventional drug and physiotherapeutic treatment aiming for increase of rehabilitation efficiency and prevention of postoperative complications in these patients.

Efficiency of combined postoperative treatment including an immunomodulatory Hexapeptide in children with acute destructive pneumonia

Acute destructive pneumonia (ADP) is a severe purulent and septic infectious disorder of childhood, characterized by a high level of morbidity and associated with imbalance of the immune system (IS). Hence, there is an obvious need to study the immunopathogenesis of this disease in order to develop new therapeutic strategy aimed at eliminating the pathogen, detoxifying the body, relieving respiratory failure and correcting functional immune deficiency. Our aim was to perform a clinical and immunological study in order to evaluate efficiency of immunomodulatory therapy using a medical drug with hexapeptide as an active substance. This drug was included into the complex postoperative treatment of children with acute destructive pneumonia. Clinical and immunological examination of 15 children 2-5 years old with ADP was performed before (study group 1 – SG1) and after (study group 1a – SG1a) combined postsurgical treatment including immunomodulatory therapy with a Hexapeptide-based pharmaceutical (HP, Arginyl-alpha-Aspartyl-Lysyl-Valyl-Tyrosyl-Arginine). Comparison group (CG) included twenty healthy children. The contents of T and B lymphocytes, natural killer cells (NK) were measured by means of flow cytometry (CYTOMICS FC 500, USA). Serum levels of IgA, IgM, IgG (ELISA), phagocytic and microbicidal activity of neutrophil granulocytes (NG) were also evaluated. Prior to the treatment in SG1 patients, a decreased number of CD3+CD19-T lymphocytes, CD3+CD8+ TCTL lymphocytes was revealed along with significant decrease in the contents of CD3-CD16+CD56+ NK (p1-3 0.05). It was found that in children with ADP, the IgG level did not differ from indices of control group, with a decrease of IgM (p1, 2 0.05), and increased level of IgA (p 0.05). We have also found a deficiency of NG effector functions, i.e., insufficiency of active phagocytic NG with impaired capture and killing of bacterial antigen. Assays of NADPH-oxidase activity showed lacking response to both inflammation and additional induction by S. aureus. After complex treatment including immunomodulatory therapy in the SG1a group, we revealed a recovery in CD3+CD19-T lymphocyte contents, CD3+CD8+TCTL lymphocytes, CD3-CD16+CD56+ NK (p1-3 0.05). The trends towards normalization of IgA, IgM, improved NG effector functions (killing ability) were revealed due to activation of NADPH-oxidases. The restoration of immunological parameters in ADP was associated with earlier recovery from the purulent-destructive process in lungs, absence of postoperative complications including the prevention of septic process. The clinical and immunological effects of the immunomodulatory therapy program with HP-based pharmaceutical preparations suggest its potential usage during postoperative period in immunocompromised children with ADP.

Correlation between the Chemiluminescent Activity of Neutrophilic Granulocytes and the Lipid Peroxidation-Antioxidant Defense System in Gastric Cancer Associated with Helicobacter pylori Infection.

To study the processes of lipid peroxidation and the activity of antioxidant defense enzymes depending on the chemiluminescent activity of neutrophilic granulocytes in patients with gastric cancer associated with H. pylori infection, depending on the stage. A total of 39 patients with stage I-II gastric cancer and 30 patients with stage III-IV gastric cancer were examined. A study of the chemiluminescent activity of neutrophilic granulocytes was carried out and the parameters of the lipid peroxidation system and antioxidant protection in plasma were determined using the spectrophotometric method. Statistical data processing was performed using the Statistica 7.0 software package (StatSoft, St Tulsa, OK, USA). In patients with gastric cancer associated with H. pylori infection, regardless of stage, the proportion of neutrophilic granulocytes with normal activity did not exceed 1/3 of the total number of patients, and the remaining 2/3 of patients had altered chemiluminescent activity of neutrophilic granulocytes. In patients with gastric cancer, by I-II stage of the disease, the majority revealed a reduced function of neutrophilic granulocytes, and in patients with gastric cancer in stage III-IV of the disease, the majority showed increased chemiluminescent activity of neutrophilic granulocytes. In all patients with gastric cancer associated with H. pylori infection, regardless of the stage of the disease, an increase in lipid peroxidation processes with activation of antioxidant defense enzymes was detected. At the same time, there were no statistically significant differences between the indicators of the system lipid peroxidation-antioxidant protection depending on the stage of gastric cancer and the chemiluminescent activity of neutrophilic granulocytes, which likely indicates that all reactive oxygen species produced by neutrophilic granulocytes in the respiratory burst are consumed locally, minimally affecting the development of oxidative stress in the blood plasma.

Glycopeptide Antibiotics Impair Neutrophil Effector Functions

Introduction: Neutrophilic granulocytes represent the first line of defense against microorganisms. Granulocytes phagocytose microorganisms and specifically synthesize oxygen radicals against them, which eventually kills the invaders. Methods: Neutrophilic granulocytes were isolated from peripheral blood of healthy volunteer donors. Putative interference of new-generation antibiotics with neutrophil function was tested using a collection of granulocyte-stimulating agents and Amplex™ Red-based plate assay and flow cytometry-based respiratory burst assays. In addition, phagocytosis of E. coli, IL-8 production, bactericidal activity, and CD62L expression of granulocytes were evaluated. Results: Of note, we found that the two glycopeptide antibiotics dalbavancin and teicoplanin inhibited ROS production upon granulocyte activation via different signaling pathways in a dose-dependent manner. Dalbavancin also blocked the PMA-induced shedding of CD62L. In contrast, the oxazolidinone antibiotics tedizolid and linezolid had no effect on neutrophil function, while the combination of ceftazidime/avibactam dose dependently inhibited the fMLP/Cytochalasin B-induced granulocyte burst in a dose-dependent manner. Additionally, we showed that dalbavancin and teicoplanin as well as sulfametrole/trimethoprim and ceftazidime/avibactam inhibited baseline and PMA-induced IL-8 production by neutrophilic granulocytes. Moreover, dalbavancin impaired the bactericidal activity of neutrophilic granulocytes. Conclusion: We here identified hitherto unknown inhibitory effects of several classes of antibiotics on the effector functions of neutrophilic granulocytes.

Analysis of leukogram parameters and blood cytokine profile in tick-borne infections of various etiologies

Introduction. Studies aimed at identifying pathogenetic features of tick-borne natural focal infections depending on etiological agent are relevant to seek out for new associations of biomarkers characterizing the structural and functional phenotype of immune cells significant for the differential diagnosis and prognosis of diseases. The aim of the study was to evaluate the parameters of a leukogram with an expanded profile in relation to the parameters of cytokine status in patients with viral tick-borne encephalitis as well as erythemic tick-borne borreliosis at acute stage of the disease. Materials and methods. The study involved 28 patients with the non-erythemic tick-borne borreliosis and 27 patients with tick-borne encephalitis at the acute stage of the disease, as well as 16 healthy individuals (control group). Venous blood samples were examined using a Sysmex XN1000 analyzer based on reflex testing technology with extended profile allowing to characterize leukocyte reaction during infection and inflammation, including parameters such as the absolute and relative count of immature granulocytes (IG), neutrophil granularity index (NEUT-GI), neutrophil reactivity index (NEUT-RI), count of reactive lymphocytes (RE-LYMP), count of antibody-producing lymphocytes (AS-LYMP). Concentrationof IL-2, IL-4, IL-6, IL-8, IL-10, IFN and TNF in blood serum was determined by enzyme immunoassay using specific reagent kits (Vector-Best, Russia). Results. It was found that changes in the quantitative composition of blood leukocytes in patients with viral tick-borne encephalitis and non-erythemic form of tick-borne borreliosis have a unidirectional tendency characterized by increased count of metabolically active neutrophils (NEUT-RI) and a decreased count of reactive lymphocytes (RE-LYMP) compared to healthy individuals. Changes in the leukogram occur along with high blood concentration of proinflammatory cytokines IL-8 and TNF and a low level of IFN. A positive direct correlation was revealed between the fluorescence intensity parameter characterizing the metabolic activity of neutrophil granulocytes (NEUT-RI) and blood serum IL-8 level both in tick-borne encephalitis (r = 0.422, p 0.05) and non-erythemic form of tick-borne borreliosis (r = 0.551, p 0.05). Additionally, in the former, a positive correlation was established between the total leukocyte count (WBC) and concentration of TNF in the blood serum (r = 0.532, p 0.05).

Composition and Biological Activity of Fetoplacental Tissues-Derived Cryoextracts Being Differently Obtained

The protein-peptide composition of fetal tissues (FTCEs) and placenta (PCE) cryoextracts of rats was investigated by gel permeation chromatography. Cryoextracts were derived from tissue homogenates using the freeze-warming modes: 1 – single (–20°С); 2 – double (–20; –196°С) and 3 – triple (–20; –196; –196°С) ones. The biological activity of cryoextracts was in vitro evaluated by the phagocytic activity of neutrophil granulocytes (NG) of the blood of intact rats after incubation with an inactivated culture of Staphylococcus aureus (2 × 109 cells/ml) for 45 and 120 min for the concentrations of extracts in the incubation medium: 1.6; 3.1; 6 mg/ml in terms of dry substance. The use of mode 3 ensured a greater yield of total protein and low molecular weight compounds of a protein-peptide nature in the composition of cryoextracts. The content of low-molecular-weight fractions in FTCE exceeded that in PCE, regardless of the obtaining mode. Incubation of NG with cryoextracts did not significantly increase the number of neutrophils which entered into phagocytosis. However, all the samples of PCE and FTCE were characterized by a dose-dependent rise in the NG absorption activity during the 45-minute incubation compared to the control.

Differences in bacteria nanomotion profiles and neutrophil nanomotion during phagocytosis.

The main goal of this work is to highlight the connection between nanomotion and the metabolic activity of living cells. We therefore monitored the nanomotion of four different clinical strains of bacteria (prokaryotes) and the bacterial phagocytosis by neutrophil granulocytes (eukaryotes). All clinical strains of bacteria, regardless of their biochemical profile, showed pronounced fluctuations. Importantly, the nature of their nanomotions was different for the different strains. Flagellated bacteria (Escherichia coli, Proteus mirabilis) showed more pronounced movements than the non-flagellated forms (Staphylococcus aureus, Klebsiella pneumoniae). The unprimed neutrophil did not cause any difference in cantilever oscillations with control. However, in the process of phagocytosis of S. aureus (metabolically active state), a significant activation of neutrophil granulocytes was observed and cell nanomotions were maintained at a high level for up to 30 min of observation. These preliminary results indicate that nanomotion seems to be specific to different bacterial species and could be used to monitor, in a label free manner, basic cellular processes.

THE EFFECT OF ALLOGENEOUS TRANSFUSION OF ERYTHROCYTE MASS ON INDICATORS OF THE FUNCTIONAL ACTIVITY OF NEUTROPHIL GRANULOCYTES IN THE BODY OF RECIPIENT RABBITS

The relevance of the study is due to the spread among animals of pathologies in the therapy of which transfusion of blood components is used, which is used both for acute blood loss and for diseases of the blood and hematopoietic organs, if their deficiency cannot be compensated by other methods. In this regard, this article is aimed at evaluating the state of phagocytic activity of phagocytic cells (neutrophils and macrophages) according to indicators of phagocytic index, phagocytic number and oxygen-dependent phagocytosis during allogeneic transfusion of erythrocyte mass in the blood of recipient rabbits. The leading approach to the study of this problem was the determination of the absorption activity of phagocytes, the determination of the oxygen-dependent bactericidal activity of blood neutrophils, which was studied using the spontaneous test with nitroblue tetrazolium (NST-test). The material for research was blood serum samples obtained from 5 clinically healthy rabbits 3, 7 and 23 days after transfusion. It was found that the phagocytic index increases by 47.7% on the 3rd day, by 34.3% on the 7th day, and by 59.1% on the 23rd day, relative to the initial state. The indicator of the phagocytic number on the 3rd day of the experiment decreased by 22.5%, on the 7th day by 21.4%, on the 23rd day by 7.2%, relative to the initial state. It was found that the metabolic activity of neutrophil granulocytes in the peripheral blood of recipient animals in the spontaneous HST test, compared to the initial state on the 3rd and 23rd day of experimental studies with transfusion of erythrocyte mass, decreases by 4.8 times. The material of the article is of practical value for the application of transfusion of erythrocyte mass in the treatment of animals with anemia of various genesis.

Effect of allogeneic blood transfusion on neutrophil functional activity and lymphocyte cytotoxicity in recipient rabbits

The relevance of this paper is that transfusion of allogeneic blood to recipient animals is always associated with immunological risks. In this regard, the purpose of this study was to assess the state of phagocytic activity of blood neutrophils by indicators of phagocytic index, phagocytic number, and oxygen-dependent bactericidal activity, as well as to establish changes in antibody-dependent cytotoxic activity of lymphocytes in recipient rabbits during allogeneic whole blood transfusion. Modelling of blood transfusions was performed on five clinically healthy rabbits by intravenous administration of whole blood at the rate of 5.5 ml/kg of body weight. Blood samples were taken from animals on Days 3, 7, and 23 after blood transfusion. Neutrophil populations were obtained from blood samples by centrifugation on a double density gradient of 1.077 and 1.093 Ficoll-Verografin. The absorption activity of phagocytes was determined in a microscopic test. To investigate the oxygen-dependent bactericidal activity of neutrophils, a spontaneous test with nitroblue tetrazolium was performed. Antibody-dependent cytotoxic activity of lymphocytes was investigated by colorimetric method. It was found that after the transfusion of whole blood, the phagocytic activity of neutrophils increases with a simultaneous decrease in their absorption capacity. On Days 3 and 7, the results of the spontaneous test with nitroblue tetrazolium decreased. This indicates inactivation of the oxygen-dependent bactericidal activity of neutrophil granulocytes during the first phase of post-transfusion immunological reactions. On Day 23, there was an increase in the values of the indicators of the spontaneous test with nitroblue tetrazolium, which indicates the activation of the bactericidal properties of phagocytes. It was found that on Day 3, the antibody-dependent cytotoxic activity of lymphocytes significantly decreased relative to the initial state, and on Days 7 and 23, it increased. An increase in the antibody-dependent cytotoxic activity of lymphocytes should be associated with the active synthesis of antibodies of the late phase of the immune response. Consequently, transfusion of allogeneic blood causes an immune response in recipient rabbits, without causing immediate and long-term transfusion reactions (changes in heart rate, respiratory rate, body temperature). The obtained results are of practical value both for scientists and practising doctors who use transfusion of whole blood and its components to animals with acute anaemia, impaired functional activity of blood coagulation factors, parasitic, and oncological diseases

Renal manifestations in vasculitides of small and medium-sized vessels

Small-vessel vasculitides, in particular, are frequently manifested in the kidneys. A distinction is made between antineutrophil cytoplasmic antibody (ANCA)-associated vasculitides (AAV) and immune complex vasculitides. Even within the AAVs there are differences with respect to renal involvement, which manifest as necrotizing glomerulonephritis (GN) but renal involvement is much rarer in eosinophilic granulomatosis with polyangiitis than in microscopic polyangiitis and granulomatosis with polyangiitis. Disease progression, organ manifestation and prognosis vary according to the ANCA status. In immune complex vasculitides (cryoglobulinemic vasculitis, IgA vasculitis, hypocomplementemic urticarial vasculitis and antiglomerular basement membrane, GBM, disease), endothelial-adjacent activation of neutrophilic granulocytes leads to local vessel wall damage with subsequent ischemic tissue damage, similar to AAV. The sparse evidence of immune complexes is different in pauci-immune AAV. Polyarteritis nodosa is adisease with variable clinical presentations with necrotizing vasculitis of small and medium-sized arteries. Intrarenal aneurysms and hemorrhages but not GN lead to renal damage. Diagnostically, the detection of specific autoantibodies (e.g. anti-GBM), cryoglobulins or increased complement turnover can be decisive. Renal biopsy with qualified immunohistopathology is particularly important in cases of initial manifestation and unclear constellation of findings. The treatment of renal vasculitis is adapted to the severity, stage of disease, extrarenal organ manifestation and pathogenesis. It ranges from glucocorticoid monotherapy to moderate immunosuppression, up to targeted biologic therapy, chemotherapy and plasmapheresis.

Whole blood transcriptome characterization of young female triathlon athletes following an endurance exercise: a pilot study.

The vast majority of studies focusing on the effects of endurance exercise on hematological parameters and leukocyte gene expression were performed in adult men, so our aim was to investigate these changes in young females. Four young (age 15.3 ± 1.3 yr) elite female athletes completed an exercise session, in which they accomplished the cycling and running disciplines of a junior triathlon race. Blood samples were taken immediately before the exercise, right after the exercise, and then 1, 2, and 7 days later. Analysis of cell counts and routine biochemical parameters were complemented by RNA sequencing (RNA-seq) to whole blood samples. The applied exercise load did not trigger remarkable changes in either cardiovascular or biochemical parameters; however, it caused a significant increase in the percentage of neutrophils and a significant reduction in the ratio of lymphocytes immediately after exercise. Furthermore, endurance exercise induced a characteristic gene expression pattern change in the blood transcriptome. Gene set enrichment analysis (GSEA) using the Reactome database revealed that the expression of genes involved in immune processes and neutrophil granulocyte activation was upregulated, whereas the expression of genes important in translation and rRNA metabolism was downregulated. Comparison of a set of immune cell gene signatures (ImSig) and our transcriptomic data identified 15 overlapping genes related to T-cell functions and involved in podosome formation and adhesion to the vessel wall. Our results suggest that RNA-seq to whole blood together with ImSig analysis are useful tools for the investigation of systemic responses to endurance exercise.

Local immune state in the women with miscarriage

Functional state of immunity provides maintenance of immunological tolerance for allogeneic fetus, and high level of local protection against antigenic stimulation. Increased functional activity of phagocytic cells at systemic and local levels may cause impairment of complete fetoplacental complex, thus leading to abortion. The purpose of our study was to assess the state of local immunity in women with pregnancy loss.&#x0D; The study involved 174 women in their reproductive age. The first group consisted of 65 patients (37.4%) with a diagnosis of miscarriage, the history of 2 abortions over the period from conception to the 20th week of gestation. The second group included 37 patients (21.2%) with documented recurrent pregnancy loss, with a history of 3 miscarriages. The control group consisted of 72 conditionally healthy women (41.4%) who had 2 or more term pregnancies with the same partner, without a history of obstetric and gynecological complications. Their mean age was 366 years old. Total number and viability of leukocytes, the indices of functional neutrophil activity, their functional reserve and neutrophil stimulation index were determined in cervical mucosal samples over the first phase of the menstrual cycle.&#x0D; In the first phase of menstrual cycle, the women with miscarriage and recurrent pregnancy loss exhibited a statistically significant increase of phagocytosis by the neutrophils from cervical mucus as compared to the group of conditionally healthy women. Intensity of neutrophil phagocytosis in cervical mucus reached higher values in the women with recurrent pregnancy loss more often, compared to the control group. Evaluation of the functional reserve of cervical mucus neutrophils in the subjects with recurrent pregnancy loss and miscarriage showed a trend towards statistically significant differences: this parameter was higher in the patients than in control group. The ability of cervical mucosal neutrophils to produce reactive oxygen species (both spontaneous and induced) did not show statistically significant differences between the patients and controls.&#x0D; Hence, we have observed aberrant functional activity of neutrophilic granulocytes from cervical mucosa in the groups of women with 2 or more abortions, without changing ability of the cells to produce reactive oxygen species. This finding may be explained by prevalence of oxygen-independent mechanisms of intracellular killing, thus suggesting a role of neutrophils for impaired balance of immunological tolerance in pregnant women at the local level.

Bioinformatics analysis of prognostic value and immune cell infiltration of SERPINA1 gene in cutaneous melanoma.

BackgroundCutaneous melanoma (CM) has a poor overall prognosis. Immune checkpoint inhibitor (ICI) therapy effectively improves overall survival in individuals with advanced melanoma, but only some patients benefit. Serpin Family A Member 1 (SERPINA1), a type of proteinase inhibitor that is used for many targets, is abnormally expressed and plays a vital role in multiple cancers. However, little is known about the clinical significance of SERPINA1 in CM.MethodsThe Cancer Genome Atlas (TCGA) and the gene expression omnibus (GEO) datasets were used to compare SERPINA1 expression levels. The association between SERPINA1 and other clinical factors were examined with R software, and receiver operating characteristic (ROC) curves for identification was developed. The Tumor IMmune Estimation Resource (TIMER) was used to examine the invasion of immune cells, markers for immune cells, and immunological checkpoints. The predictive value of SERPINA1 DNA methylation levels for every CpG was analyzed with the MethSurv web tool. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used to assess the roles of genes that interacted with SERPINA1. The Tumor Immune Dysfunction and Exclusion (TIDE) algorithm was used to predict SERPINA1’s response to ICIs.ResultsSERPINA1 was substantially expressed in CM. Overexpression of SERPINA1 was significantly associated with CM severity. The outcome for individuals with elevated SERPINA1 expression was good (HR =0.54, P<0.001). Using SERPINA1 expression levels, tumors and normal tissues could be reliably differentiated [area under the curve (AUC) =0.889]. Positive associations were found between SERPINA1 in CM and the infiltration of immune cells and immunological checkpoints [programmed cell death-1 (PD-1) and CTLA-4]. The efficacy of immune checkpoint blockade (ICB) in patients with a low expression of SERPINA1 was good. The GO pathway enrichment analysis showed that activation of neutrophil granulocytes participated in enrichment in the immune response pathway. Patients with low SERPINA1 expression had low TIDE scores.ConclusionsSERPINA1 is involved not only in the development and progression of CM but also in the immunological control of CM. Thus, SERPINA1 may serve as a possible biomarker for CM diagnosis, as well as its therapeutic target.

The role of immunological factors in the development of abnormal uterine bleeding in women of reproductive age with extragenital disorders

The aim of the study was to assess the role of immunological factors in the development of abnormal uterine bleeding in women of reproductive age with extragenital disorders.&#x0D; Materials and methods. The study involved 100 women with abnormal uterine bleeding and accompanying extragenital disorders (main group) and 50 somatically healthy women (control group). Autoimmune antibodies to platelets, phagocytic activity of neutrophil granulocytes, concentration of circulating immune complexes (CICs), total level of membranotropic cytotoxic factors, content of CD4+T-helper subpopulations and cytotoxic CD8+T-killer lymphocytes were evaluated as immunological markers.&#x0D; Results of the study. The study showed that thrombocytopenia, caused by the presence of autoimmune antibodies to their own platelets, can be one of the pathogenic factors of bleeding in women with AUB. In 41 % of women with AUB, phagocytic reactions were found to be intense, which was expressed by an increase in chemotaxis and adhesion functions, and in 46 % of women by an increase in the absorption capacity of phagocytes. In the main group, 48 % of the examined women had insufficient phagocyte enzymatic activity, which was evidenced by a decrease in the index of completion of phagocytosis. In 79 % of women of the main group, violations of the formation and elimination of circulating immune complexes were detected. The formation of low-molecular-weight CICs in 82 % of women of this cohort contributed to the induction of autoimmune reactions. The total content of membranotropic cytotoxic factors, which was evaluated according to the lymphocytotoxic test, exceeded the reference values in 88 % of women of the main group. In the main group, the average content of CD4+ T-helpers was 23 % lower, and the content of suppressor CD8+ T-lymphocytes was twice as low compared to the control group, resulting in a significant increase in the immunoregulatory index by 30 %.&#x0D; Conclusion. The women of the main group with abnormal uterine bleeding were found to have a violation of the functional activity of cellular factors of innate immunity, accompanied by changes in the absorption and digestive capacity of phagocytic cells. Assessment of secondary adaptive reactions showed induction of humoral sensitization and formation of autoimmune reactions (presence of antiplatelet autoantibodies, increase in CICs and LCT, decrease in the subpopulation of CD8+-suppressor T-lymphocytes). The detected violations indicate the pathogenic role of immunological reactions in women with abnormal uterine bleeding

Phagocyte Activity in Response to Exposure to Methicillin-Resistant &lt;i&gt;Staphylococcus aureus&lt;/i&gt; Strains

The research focuses on the study of phagocyte activity of blood neutrophilic granulocytes, monocytes, and eosinophils under the exposure to methicillin-resistant strains of Staphylococcus aureus bacteria. The subjects of the research were represented by blood neutrophilic granulocytes, monocytes, and eosinophils, isolated from healthy people, as well as methicillin-resistant and methicillin-sensitive S.aureus bacteria strains (MRSA and MSSA). Phagocytosis functions (phagocytic number and phagocytic index) were estimated by FITC-labeled bacteria. The analysis of stained cells was performed using FC-500 flow cytofluorimeter (Beckman Coulter, USA) for whole peripheral blood. Phagocytosis research resulted in the following findings. In response to MRSA, the percentage of neutrophils involved in phagocytosis and average number of bacteria being present inside the cells increase as compared to those within sensitive strains. In response to MRSA exposure, the phagocytic index is higher in monocytes with CD14+CD16¯phenotype. At the same time, phagocytic number is higher in non-classic populations of monocytes CD14+CD16+and CD14lowCD16+. Moreover, it should be noted that the classical type of monocytes CD14+CD16¯tends to be activated faster in regard to MRSA, but phagocytosis efficiency is lowered. Blood eosinophils also actively respond to MRSA. Thus, changes in functional activity of neutrophilic granulocytes, monocytes, and eosinophils in peripheral blood were established during MRSA induction. The bacterial receptor apparatus changes due to the modification of cellular wall as a result of acquired resistance to antibiotics.

A complex diagnostics of early gastric cancer associated with &lt;i&gt;Helicobacter pylori&lt;/i&gt; infection in the adult population of the Krasnoyarsk Territory

The aim of our study was to evaluate the clinical-anamnestic, serological, immunological and biochemical tests used for early diagnostics of gastric cancer associated with Helicobacter pylori infection in the adult population of the Krasnoyarsk Territory. Materials and methods. The control group consisted of 104 apparently healthy blood donors, the comparison group – 97 patients with chronic atrophic gastritis as well as a group of patients with early gastric cancer comprising 98 subjects. Assessment of monocyte and neutrophil spontaneous and induced chemiluminescence (CL) was carried out on a 36-channel biochemiluminometer BLM-3607. Phagocytosis was measured by using a Beckman Coulter FC 500 flow cytometer. A Varian Cary Eclipse spectrofluorometer was used to study lipid peroxidation and factors of the antioxidant defense system. Results and discussion. While studying the phagocytic arm of immunity, it was found that all patients with early gastric cancer were reported to have parameters of the maximum intensity for neutrophil spontaneous CL from 17 831 c.u. and lower, whereas induced CL reached at least 30 000 c.u. Phagocytic activity of neutrophilic granulocytes in patients with early gastric cancer was 36% or less. While studying the indicators of monocytes, it was found that spontaneous and induced CL decreased from 454 c.u. and 1186 c.u., respectively, in the patients with early gastric cancer. Monocytic activity in early gastric cancer was 34% or less. In the study of lipid peroxidation, an antioxidant defense in patients with chronic atrophic gastritis and gastric cancer had increased malondialdehyde (MDA) level. Patients with gastric cancer had decreased activity of the enzyme catalase (CAT), whereas subjects with chronic atrophic gastritis had reduced glutathione peroxidase (GPO) level. In contrast, patients with early gastric cancer were featured with increased GPO activity. We have proposed coefficients for assessing the factors of the antioxydant defence system in patients: the ratio for superoxide dismutase to catalase activity (SOD/CAT) as well as the ratio for superoxide dismutase to glutathione peroxidase activity (SOD/GPO). Conclusion. During the study, threshold values of parameters were obtained for assigning groups at high risk of developing early gastric cancer, which can be used for screening in adult population.

The role of T cells and interleukin‐17A in Behçet disease

British Journal of DermatologyVolume 185, Issue 6 p. e201-e201 Plain Language Summary The role of T cells and interleukin-17A in Behçet disease First published: 17 December 2021 https://doi.org/10.1111/bjd.20788AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Abstract Linked Article: Vural et al. Br J Dermatol 2021; 185:1209–1220. Behçet disease (BD) is a severe chronic inflammatory disease. It damages blood vessels and can affect all organs, including the skin, eyes and brain. This can lead to blindness or even death. BD is common along the historical ‘Silk Road’, extending from Europe to the Far East, especially in Turkey. To find out what causes the disease, researchers from Turkey, Germany and Japan investigated skin lesions from patients with BD. BD is associated with the genes for HLA-B*51, ERAP1 and the interleukin (IL)-23 receptor. These genes determine targets of CD8+ T-cell responses and the production of the immune mediator IL-17. High levels of IL-17 in the blood are characteristic of BD. In psoriasis, another inflammatory skin disease, an HLA-class I molecule, together with ERAP1, induces an autoimmune response (where the body’s immune system attacks its own healthy cells and tissues) and high IL-17 production by CD8+ T cells against the pigment-producing cells of the skin, the melanocytes. This suggests a similar autoimmune disease pathway in BD. We found that in skin lesions from patients with BD activated CD8+ T cells producing high levels of IL-17 induced by IL-23 prefer to accumulate around blood vessels. This causes recruitment and activation of neutrophilic granulocytes, which contribute to tissue damage by forming networks of released DNA. These findings propose that in BD dysregulated CD8+ T cells wrongly attack the body’s own blood vessels. This study improves our understanding of how these genetic traits contribute to a disease pathway, which may help to develop new and effective treatments for BD targeting IL-17 or IL-23. Volume185, Issue6December 2021Pages e201-e201 RelatedInformation