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The results of recent studies show that tumor biology, coagulation activation, and inflammatory reactions profoundly contribute to the thrombosis pathogenesis in cancer as well as tumor progression, metastasis, and developing chemoresistance. Cancer is an independent predictor of thrombosis. During carcinogenesis, tumor cells express proinflammatory cytokines, proangiogenic and procoagulant factors, and also stimulate other cells to express various components promoting emerging thromboinflammation. The discovery of neutrophil extracellular traps (NETs) provides an opportunity to take a new look at biology and a role neutrophils may play in thromboinflammation and tumorigenesis. The close interplay between tumor cells, tumor-associated neutrophils and NETs as well as other players in the tumor microenvironment underlies activation of thromboinflammation in cancer patients not only resulting in thrombus formation, but also promoting tumor growth and dissemination.

Scientific and technological progress contributes to the discovery and production of innovative materials. The emergence of graphene is a clear example of this. Graphene is considered a promising material for use in nanobiomedicine and nanobiotechnology. It is therefore important to understand how it affects human immune cells. In a study, the effects of 5 and 25 μg/mL graphene oxide nanoparticles with lateral sizes of 100-200 nm and 1-5 μm, modified with linear and branched polyethylene glycol, on human neutrophils were investigated. The formation of reactive oxygen species was evaluated with a lucigenin as a chemiluminescence activator.Inaddition, we investigated theeffect of a 60-minute incubation of neutrophils with pegylated graphene oxide nanoparticles on the viability of these cells by staining with trypan blue and a 30-minute incubation on the uptake of fluorescein isocyanate-labelled E. coli. The percentage of neutrophils which engulfed E. coli and the uptake index were determined. Samples without added nanoparticles served as controls.A decrease in lucigenin-enhanced chemiluminescence of neutrophils was observed under the influence of two types of graphene oxide nanoparticles: 1-5 μm in size coated with linear polyethylene glycol, and 100-200 nm in size coated with branched polyethylene glycol, at a concentration of 25 μg/mL in the zymosan-stimulated version of the assay. No dependence of the effect on the particle size and the type of polyethylene glycol was observed. The indicators for spontaneous chemiluminescence of neutrophils did not change with the addition of PEGylated graphene oxide nanoparticles.A thirty-minute incubation of human neutrophils at 37 °C with PEGylated graphene oxide nanoparticles with lateral dimensions of 100-200 nm and 1-5 μm had no effect on the viability of these cells and on the percentage of neutrophils that engulfed E. coli. However, 1-5 μm graphene oxide modified with linear polyethylene glycol at a concentration of 25 μg/mL increased the amount of E. coli engulfed by neutrophils per cell.Thus, in the absence of cytotoxicity, PEGylated graphene oxide particles have multidirectional immunomodulatory effects on neutrophils. In this case, their concentration is decisive and not the size of the graphene oxide particles and the type of polyethylene glycol.

Antineutrophil cytoplasmic antibody-associated systemic vasculitis (ANCA-SV) is a group of rare and potentially severe systemic diseases. The search for reliable methods to assess ANCA-SV activity remains relevant. Among the indicators of neutrophil activation that have emerged in clinical practice, the level of serum calprotectin (CLP) stands out, which can be a marker for monitoring vasculitis activity and identifying patients at risk of disease relapse. Objective: to determine serum CLP levels in patients with ANCA-SV. Material and methods. The study group comprised 64 patients (37 with granulomatosis with polyangiitis, 11 with eosinophilic granulomatosis with polyangiitis and 16 with microscopic polyangiitis) aged 18 years and older with a confirmed diagnosis of ANCA-SV. The control group consisted of 30 healthy individuals. ANCA-SV activity was determined using the BVAS index; high activity corresponded to a BVAS value of >3. Damage was assessed using the VDI index. Depending on ANCA-SV activity, patients were divided into two groups: high activity group (group 1, n=33) and low activity group (group 2, n=31). In addition to the generally accepted indicators, serum CLP levels were assessed in all patients with ANCA-SV and healthy donors. Results and discussion. Statistically significant differences (p

The purpose of this study was to investigate the effect of neutrophils and their antimicrobial factors, hydrogen peroxide and defensin α, on the biofilm biomass, the viability of bacteria in the biofilm and the efficiency of conjugative transfer of the pOX38:Cm plasmid from the E. coli N4i pOX38:Cm strain into different E. coli strains (commensal К12 TG1 and uropathogenic DL82, R32 and R45). The biofilm of the recipient E. coli TG1 with the donor E. coli N4i pOX38:Cm increased when 10⁵ cells/ml of neutrophils were added compared to the control, while the biofilm biomass of the uropathogenic E. coli recipient strains DL82/E. coli R45 with the donor E. coli N4i pOX38:Cm decreased when 10⁶/10⁴–10⁶ cells/ml of neutrophils were added, respectively. The survival of recipient E. coli TG1 cells and transconjugants in the biofilm was, compared to the control, higher when 10⁴, 10⁵, 10⁶ cells/ml of neutrophils were added. The addition of 0.1 mM H₂O₂ increased biofilm formation of E. coli DL82 and E. coli R45, and addition of 0.5 mM H₂O₂ reduced biofilm formation of E. coli DL82, while 0.5 mM or 2.5 mM reduced the E. coli R45 bacterial biofilm biomass in the conjugative mixture. The frequency of the pOX38:Cm conjugative transfer was lower in the presence of 2.5 mM H₂O₂ in the N4i pOX38:Cm × DL82 biofilm, and also in the presence of 0.5 and 2.5 mM H₂O₂ in the N4i pOX38:Cm × R45 biofilm, compared to the control. The frequency of pOX38:Cm conjugation from the donor E. coli N4i pOX38:Cm into E. coli DL82 decreased, when 5 or 25 ng/ml defensin α were added and the conjugation frequency in the mating mixture N4i pOX38:Cm×R45 decreased, when 5 ng/ml were added, while, when 25 ng/ml of defensin α were added it increased.

The paper reports on the research into the immunotherapeutic properties of a conjugate based on BCG antigens with betulonic acid after experimental infection of guinea pigs with Mycobacterium scrofulaceum culture, belonging to nontuberculosis mycobacteria type II according to the Runyon classification. Fifteen guinea pigs were used for the experimental purposes, divided into 3 groups. Mycobacterium scrofulaceum was subcutaneously injected into animals of Groups 1 and 2 (n = 10) at a dose of 5 mg. Fourteen days later, a conjugate based on BCG antigens with betulonic acid was subcutaneously injected into animals of Group 2 (n = 5) at a dose of 500 µg/mL of protein. Five intact animals were used as controls. During the experiment, neutrophil bactericidal activity was assessed, and histopathological examination of inguinal lymph nodes was done. The experiment showed that the inoculation of Mycobacterium scrofulaceum into guinea pigs activates cationic proteins and neutrophil myeloperoxidase, and on experiment day 42 (preceded by mycobacteria withdrawal from the body) their concentration reduced to the level of the control group. The vaccine administration induced a more active intracellular phagocyte metabolism during the entire observation period, which resulted in the elimination of nontuberculosis mycobacteria in animals as early as day 7 after treatment with the conjugate. The elimination was confirmed by the absence of mycobacterial antigen in blood smears tested in indirect immunofluorescence, as well as by histopathological changes in inguinal lymph nodes demonstrated as a reduction of germinal centers within lymphoid follicles.

Asthma, caused by eosinophilic inflammation of the airways, is caused by exposure to allergen, is well treated with glucocorticostroids or monoclonal antibodies to interleukin-4 and 5, but some patients do not develop an answer to this therapy. The given type of asthma is classified as non-eosinophilic asthma. Depending on the proportion of infiltrating cells, non-eosinophilic asthma can be divided into neutrophil asthma (NA), mixed granulocytic asthma, and small ranulocytic asthma. It should be noted that the NA criteria are not clearly defined. For example, in one study, inflammation is thought to be neutrophilic with less than 2.5% of eosinophils and more than 65% of neutrophils in the sputum, while in another study 61% of neutrophils and more are thought to be. The role of neutrophils in asthma has been studied, but there is debate about the presence of NA. Several studies have shown that neutrophilic inflammation is associated with severe asthma and the risk of increased asthma. The article considers pathogenesis, definition, biomarkers NA and potential therapy NA.

Aim. To assess the prognostic value of the formation of neutrophil extracellular traps (NETs) in blood smears of operated patients with infective endocarditis (IE).Material and methods. We prospectively included 46 patients with verified IE and 50 patients with valvular heart disease without IE, hospitalized in a cardiac surgery hospital in 2021-2022 (Moscow), comparable by sex and age. In all patients, NETs were determined upon admission and 7 days after surgery using the MECOS-Ts2 automated microscope (Russia). Patients included in the study were prospectively followed during the in-hospital period (ME [IQR] 30,0 [21,0-41,0] days) for the primary composite endpoint (in-hospital all-cause mortality, embolic, intracardiac, infectious complications) and its individual components.Results. Patients with IE were predominantly male (n=37, 80,4%) with a median age of 55,5 [44,0-70,0] years. The primary combined endpoint was recorded in 76,1% (n=35) of those examined with IE. The NET level after 7 days was significantly higher in patients with IE who had primary combined endpoint events than in the group of operated patients with heart defects without IE (Me [IQR] 4,4 [0,6-26,6] vs 2,9 [1,1-4,3], respectively, p

Background. Intensive milk production technology creates a number of factors that can put an animal into a state of technological stress. The effect of stress is reflected in nonspecific resistance, which represents the body's ability to resist the effects of adverse factors by maintaining protective forces at the proper level. Neutrophils are one of the first protective cellular barriers that are rapidly mobilized when exposed to stress factors. To decipher the changes taking place in response to the action of stressors, the involvement of various evaluation tools is required. One of the possible methodological approaches is the use of various visualization methods that provide access to spatio-temporal information at the level of individual organelles and Purpose. The study of the morpho-functional state of neutrophils under technological stress in cattle by laser interference and electron scanning microscopy. Materials and methods. The work was carried out on 30 clinically healthy highly productive black-and-white cows. A combination of impacts was considered as technological stress: the rearrangement of animals and the change of service personnel. Neutrophils were analyzed by laser interference and electron scanning microscopy. Results. A change in the morphological structure and functional activity of neutrophils after the action of technological stress is shown. The analysis of interferograms revealed an increase in the heterogeneity of neutrophils due to the appearance of degeneratively altered cell forms with an increase in the number of functionally active neutrophils. Electron scanning microscopy revealed the appearance of NEToses and neutrophils with emptied granules and vacuolization of the cytoplasm after technological stress. Conclusion. The revealed morphofunctional changes in neutrophils prove the strain of adaptation mechanisms under technological stress. This circumstance must be taken into account in order to prevent disruption of the adaptive capabilities of the organism of highly productive animals.

Aim. To evaluate the diagnostic and prognostic value of circulating mature and aging neutrophils in relation to hypoechoic carotid plaques and short-term progression of carotid and multifocal atherosclerosis.Material and methods. The study included 200 patients (89 males and 111 fe­males), aged 40-64 years. All patients underwent duplex ultrasound of the carotid and lower extremity arteries at the first visit and at a repeat visit after 12-24 months. Ultrasound morphology of carotid plaques was assessed using greyscale median analysis. Phenotyping and differentiation of neutrophil subpopulations was carried out using flow cytometry.Results. The absolute and relative number of mature neutrophils directly correlated with ultrasound indicators of carotid atherosclerosis, while the number of aging neutrophils — with the degree of lower extremity artery stenosis. Patients with hypoechoic carotid plaques were characterized by a significantly higher absolute number of mature neutrophils (p=0,0340). An increase in the number of mature neutrophils over 3023,0 cells/μL made it possible to predict the hypoechoic carotid plaques with a sensitivity of 75,0% and a specificity of 69,5%. Patients with carotid atherosclerosis progression had a higher absolute number of mature neutrophils (p=0,0140), as did patients with progression of multifocal atherosclerosis (p=0,0162). An increase in the number of mature neutrophils more than 3223,0 cells/μL was associated with an increase in the relative risk of polyvascular disease progression by 3,09 times (95% confidence interval, 1,34-7,17; p=0,0082) after adjustment for baseline cardiovascular disease risk.Conclusion. Among patients aged 40-64 years, increased numbers of circulating mature neutrophils are associated with an increased carotid plaque burden and hypoechoic carotid plaques. An increase in the number of mature neutrophils over 3223,0 cells/μL was associated with a 3,09-fold increase in the relative risk of polyvascular disease after adjustment for baseline cardiovascular risk.

COVID-19 is an infectious disease that ranges from the common cold to severe interstitial pneumonia, respiratory failure and septic shock. Patients with severe COVID-19 are characterized by a dysfunctional immune response, whereby neutrophils may amplify pathological damage or control other cell subsets. Despite the rapid study of all parts of the immune system in COVID-19 patients, there remain many unknown critical features of neutrophils in the control of viral infections. This article reviews the role of neutrophils in the pathogenesis of COVID-19 with emphasis on the release of neutrophil traps, neutrophil heterogeneity, and mechanisms involved in immunothrombosis. The knowledge summarized in this study should help integrate neutrophil biology to develop new and more effective COVID-19 therapies.

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

The similarity of clinical manifestations and some pathogenetic mechanisms in rheumatoid arthritis (RA) and osteoarthritis (OA) is of particular interest in studying the features of extracellular neutrophil traps (NETs) formation in these musculoskeletal diseases. Our objective was to evaluate circulating neutrophil extracellular trap formation related to autoimmune inflammation in RA and reactive inflammation in OA.39 RA patients comprised the main group; 35 OA patients, control group; 33 healthy individuals were included into the reference group. Circulating neutrophils were isolated by the one-step centrifugation in double layer of iohexol density gradient. NETs were induced by phorbol-12-myristate-13–acetate (PMA) being visualized by fluorescence microscopy.The RA disease activity at the inclusion period should not exceed 2.6 DAS28 points. ОA patients were in clinical remission at the inclusion timepoint. In 17 RA patients, an increase in DAS28 exceeded 3.2 points; in 25 OA patients, an exacerbation was diagnosed during the study. The mean percentage of spontaneous and induced NETs in reference group was 3.8% (2.6-5.0) and 12.2% (9.0-15.4), respectively. Appropriate values for the patients with inactive RA were 5.9% (5.6-6.2) and 26% (23.9-28.1); 16.6% (16.1-17.1) and 38.0% (36.6-39.4) in active RA cases, respectively; 5.4% (5.2-5.6) and 20.3% (18.3-22.3) in OA without synovitis; 13.1% (12.6-18.1) and 28.3% (6. 4-30.2) in OA with synovitis, respectively. Spontaneous increase of NETs during RA activation was 181.4%; induced increase in NETs was 46.2%; and in cases of OA exacerbation these values were 142.6% and 39.4%, respectively. In RA patients, the rates of spontaneous NETs formation were 3.9 times higher than for induced values. Appropriate index was 3.6 times higher among the OA patients. The increase in spontaneous -to-induced NETs formation was more pronounced in active RA than in OA with synovitis. The main difference in NETs composition for active RA patients (in 88% of cases) and OA synovitis (in 50% of cases) is related to the contents of citrulline epitopes (p = 0.03).Transition of RA from the remission state to active inflammation, as well as OA exacerbation are accompanied by a significant increase in NETs, especially, spontaneous formation. The degree of increase in RA was higher than in OA, thus, probably, indicating higher involvement of neutrophils in generation of NETs during autoimmune inflammation rather than in reactive inflammation. A sufficient role of citrullinated epitopes in NETs demonstrates their influence upon induction and maintenance of autoimmune response to RA-specific autoantigens.

Polyphenols exert a wide range of biological effects, including immunomodulatory action. Studying the effects of flavonoids on phagocytic activity of specialized phagocytic cells seems to be a rather promising direction for their further usage as pharmacological (therapeutic) agents. Quercetin and luteolin are the most commonly studied flavonoid substances with pleiotropic action. In-depth study and understanding of immunotropic mechanisms (e.g., regulation of phagocytosis) is a prerequisite for adequate pharmacotherapy in infectious conditions, nonspecific inflammatory diseases, autoimmune and oncological disorders. The aim of our work is to study the effect of flavonoids upon phagocytic activity of professional phagocytes (neutrophils) using an in vitro test system. The biological material (venous blood) from 30 practically healthy people (adults n = 15, children n = 15) was used in the present work. The study was carried out in accordance with established international regulations. 0.5 mg/L of Luteolin (basic substance content ≥ 98%) and Quercetin (basic substance content ≥ 95%) were added to experimental samples and incubated for 20 min at 37 °С. The percentage of phagocytosis, phagocytic number (mean number of formalin-treated sheep erythrocytes engulfed per one neutrophil) was determined in control and experimental samples using light microscopy. The unidirectional nature of phagocytosis inhibition by quercetin and luteolin was noted in the test experiments. A statistically significant (p < 0.05) decrease in the phagocytosis intensity by 10% was shown in experimental blood samples obtained from adult patients compared with control values. When quercetin and luteolin were added to blood samples obtained from children, a statistically significant (p < 0.05) decrease in phagocytosis by 30% was noted against control values. At the same time, the mean percentage of phagocytosis and phagocytic number in blood samples after the addition of flavonoids were found to be in the range of reference values, thus suggesting adequacy and physiological suppression of excessive activities of innate immunity compartments by quercetin and luteolin. At this concentration, the flavonoids were found to exert a more pronounced suppressive effect on phagocytic activity in children. Modeling of immune response using the phagocytosis indices assayed in experimental in vitro models with neutrophils from practically healthy adults and children enables us to expand the knowledge of mechanisms underlying the immunotropic effects of flavonoids (quercetin and luteolin), in order to correct immunopathological conditions.

Hematopoietic stem cell transplantation is used to treat hemoblastoses and some other diseases. Depending on the diagnosis, autologous cells of the patient or allogeneic cells obtained from a healthy related or unrelated donor are used. A sufficient count of harvested hematopoietic progenitor cells is necessary for successful transplantation. Currently, stimulation of their egress from the bone marrow into the peripheral blood by a granulocyte colony-stimulating factor, followed by collection by leukapheresis, is widely used for their preparation. Hematopoietic stem cells leave the bone marrow niche formed by the stromal microenvironment when their holding bonds are interrupted. Mesenchymal stromal cells regulate the release of hematopoietic precursors by producing various cytokines and other biologically active substances. Therefore, the study of the functional properties of bone marrow mesenchymal cells can help in solving the problem of “poor” mobilizations that occur in patients with multiple myeloma.The aim of the study was to estimate the effectiveness of mobilization of hematopoietic stem cells depending on the cytokine-producing ability of mesenchymal stromal cells in the bone marrow of donors and patients with multiple myeloma.The yields of hematopoietic progenitors were studied in 10 donors (median age 27 years) and 18 patients with multiple myeloma (median age 57 years). In donors, the release of hematopoietic stem cells into the peripheral blood was stimulated by subcutaneous administration of G-CSF at a dose of 10 μg/kg/day. Patients with multiple myeloma received vinorelbine at a dose of 35 mg/m2 of body surface, followed by administration of G-CSF (10 μg/kg/day subcutaneously). A culture of mesenchymal stromal cells was obtained from bone marrow taken prior to mobilization and the content of interleukins IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-α and IFN-γ in supernatants was studied by enzyme-linked immunosorbent assay using kits of reagents produced by Vector-Best (Novosibirsk).Patients with multiple myeloma were characterized by high secretion of the pro-inflammatory cytokine IL-2 (4.70 pg/ml vs 3.55 pg/ml in healthy individuals, p=0.003) and low IFN-γ (0.41 pg/ml vs 2.14 pg/ml in healthy, p

Purpose of the study. To carry out a comparative analysis of lipid peroxidation intensity and antioxidant system indices in blood neutrophils of patients with primary and relapsed soft tissue sarcomas (STS) depending on sex and age.Patients and methods. Of the 81 patients included in the study, 48 had primary STS, 5 patients with continued growth, and 28 patients with recurrent STS. The patients were divided by sex and age; the level of lipid peroxidation products, superoxide dismutase (SOD) activity, total peroxidase activity (TPA), glutathione peroxidase (GPx) activity and reduced glutathione (RG) content in blood neutrophils were investigated by conventional spectrophotometric methods. The comparison groups (donors) consisted of 12 men and 17 women divided into the same age subgroups: ≤ 45 years and > 45 years.Results. In donors, there was a multiple decrease in the SOD/TPA ratio with age, especially in women, and in men this was accompanied by an age-related decrease in GPx activity. In primary sarcomas in older women and in men with continued growth and recurrence, there was an increase in diene conjugates (DC). In men over 45 years of age, there was an increase in SOD, TPA, and GPx. Women of the older age group were characterized by activation of SOD and GPx, expressed in the relapsed process to a significantly greater extent than in men, and an increase in RG was observed in women. The increase in both components of the glutathione system and SOD activity was especially significant in the development of relapses in women for periods exceeding three years, which was accompanied by a decrease in DC content.Conclusions. Neutrophils are characterized by an increase in DC content in patients of both sexes in the older age group with primary and recurrent STS, with its highest content in men with continued growth and relapses. GPx activity, increased in both men and women in the older age group in all variants of STS development, may play an important role in the antioxidant protection of blood cells in STS. Thus, the most pronounced activation of GPx, accompanied by a maximum increase in reduced glutathione and activation of SOD, contributes to a decrease in the level of DC and the absence of MDA increase in women with slow development of recurrences.

Uncontrolled activation of neutrophils is considered an important mechanism of thromboinflammation and fibrosis in immunemediated rheumatic diseases (IMRD), malignant neoplasms, atherosclerosis, COVID-19 and many other acute and chronic inflammatory diseases of humans. Particular attention has been drawn to the ability of neutrophils to form “network” (web-like) structures, called “neutrophil extracellular traps” NETs. The process associated with the formation of NETs and the weakening of their degradation is called “NETosis”. The publication summarizes data on the role of NETosis in the pathogenesis of IMRD and discusses the prospects for pharmacotherapy aimed at preventing the formation and destruction of NETs.

Background. The intensification of animal husbandry leads to a violation of the adaptive capabilities of the body, a decrease in productivity and natural resistance of animals. The study of nonspecific factors of protection of the cow's body will allow us to develop ways to increase the immune status and as additional information can be used in breeding work for the selection of highly resistant parent pairs. Purpose. The study of the cellular and humoral link of nonspecific resistance in the development of a stress reaction in cattle after the action of technological stress on the body of cows. Materials and methods. The work was carried out on 20 clinically healthy highly productive black-and-white cows. The stressor for the cows was the regrouping, the change of attendants, the conduct of veterinary and sanitary manipulations. The total number of leukocytes, leukogram, bactericidal and lysozyme activity of the blood serum of cows were studied by standard clinical and laboratory methods and the morphology of leukocytes on a scanning electron microscope before and 3, 14, 30 days after technological stress. Results. On days 3-14 after stress, leukocytosis, neutrophilia, monocytosis and lymphopenia were detected relative to the initial level. When using high-resolution microscopy using a scanning electron microscope, the appearance of NEToses in the blood of cows after technological stress was detected. A decrease in the bactericidal activity of serum on the 3rd day after stress with its subsequent increase was established, on the contrary, an increase in lysozyme on the 3rd day was accompanied by its subsequent decrease by 30 days after the technological stress relative to the initial level. Conclusion. The data obtained indicate that the body's defenses are a dynamic physiological indicator, which must be considered as a general resistance of the cattle body to stressors in order to prevent the disruption of the body's adaptive capabilities.

The role of neutrophils in kidney cancer is currently being studied. Their role in carcinogenesis is ambiguous. As one of the most abundant blood leukocytes, neutrophils play an important role in cancer progression through multiple mechanisms, including promotion of angiogenesis, immunosuppression, and cancer metastasis. Neutrophils synthesize and release pro-angiogenic factors that are able to directly or indirectly stimulate the growth and migration of endothelial cells, which in turn causes the formation of new blood vessels from pre-existing ones. The production of various factors by neutrophils, including proangiogenic ones, is mediated by the expression of the genes of these molecules. Functional heterogeneity is characterized by differences in neutrophil gene expression patterns. The aim of this study was to evaluate the angiogenic potential of circulating neutrophils in kidney cancer. The object of the study were blood neutrophils of patients with verified clear cell kidney cancer at stage I (T1N0M0G1, n = 28, median age 60), stage II (T2N0M0G2, n = 15, median age 61) and stage III (T3N0M0G2, n = 15, median age 63) before surgery. The control group consisted of apparently healthy donors (n = 15, median age 54). Serum levels of IL-8 and VEGF-A were assessed by enzyme immunoassay. Expression of the CXCL8 and VEGF-A genes in circulating neutrophils was determined by reverse transcription quantitative PCR. As a result of our study, an increase in the level of IL-8 and VEGF-A in the blood serum of patients with kidney cancer in all studied groups compared with the control group was revealed. We observed a direct correlation between serum levels of IL-8 and VEGF-A in patients with kidney cancer (r = 0.429; p = 0.016), which confirms the relationship of these angiogenic factors. A significant increase in CXCL8 gene expression by circulating neutrophils was found in patients on II (2.91, Q0.25-Q0.75: (1.296-4.99), p = 0.02) and III (1.93, Q0.25-Q0.75: (0.755-11.36, p = 0.014) stages of kidney cancer compared with the control group (1.50, Q0.25-Q0.75: (0.80-4.05)). However, VEGF-A gene expression by circulating neutrophils did not differ from those in the control group. Blood neutrophils in kidney cancer exercise their angiogenic potential through the production of IL-8.

The transition of cervical neoplasia (CIN) to cervical cancer occurs with the active participation of IL-4, for which both pro- and antitumor effects have been shown with tumors of various localizations. The expression of cytokines is regulated at the transcriptional level in the promoter region of the gene. It has been shown that the genotype IL4 (589C/T) (rs2243250) is associated with the development of gastric and breast cancer. The contribution of IL-4 genotypic variations to the development of CIN has not yet been studied. The aim of the study was to assess the risk of developing cervical neoplasia by the presence polymorphism of IL4 (589C/T) and the level of IL-4. The object of the study was circulating neutrophils, serum and genomic DNA of 36 patients with CIN and 20 women without dysplasia (comparison group). Using ELISA, the level of IL-4 was determined in neutrophil lysate and serum. Phagocytic activity and adhesive ability (CD11b) of neutrophils were assessed. Allele-specific real-time PCR using Taq-Man probes was used to analyze of the IL4 589C/T (rs2243250). Statistical processing was carried out using Statistica 13 and Jamovi 1.6.5.0. As a result of the study, it was found that the level of IL-4 in serum and circulating neutrophils in patients with CIN is significantly higher than in the comparison group. The -589C* allele of the IL4 gene and the TT genotype are more common in the group with CIN (55.5%) than in the control (25%). At the same time, a direct relationship was established between the presence of polymorphism and increased adhesive ability and with indicators of the phagocytic number of circulating neutrophils. Analysis of the incidence of IL4 C589T by the «case-control» method showed that the chances of CIN formation in carriers of the -589C allele and the TT genotype were 3.75 (95% CI: 1.013 - 13.880, Chi-square = 4.161, p = 0.042). The -589C* allele and TT IL4 genotype, neutrophil and serum IL-4 levels are associated with HPV infection. Using a binary logistic regression model, we demonstrated the possibility of using IL-4 levels in circulating neutrophils and IL-4 gene polymorphism (589C/T) for the differential diagnosis of patients with CIN (χ2 = 15.6, p = 0.001). Significant significance for their combination was assessed by ROC-curve analysis (IL-4 in neutrophils; IL4 (-589С*), 75% probability. Thus, the IL4 (589C/T) is associated with the adhesive and phagocytic activity of circulating neutrophils. In HPV-infected patients, IL4 gene polymorphism (589C/T) can serve as a marker for early detection and prognosis of CIN.

Bruton's tyrosine kinase (BTK) inhibitors represent a class of drugs that have demonstrated their efficacy and safety in patients with chronic lymphocytic leukemia and non-Hodgkin's lymphomas who were considered refractory to any previously used type of therapy. BTK plays a key role in all stages of B lymphocyte development, but in recent years, there have been data indicating that BTK is also involved in the activation of myeloid cells.The aim of this study is to analyze and systematize all published materials on the immunomodulatory effects of BTK inhibitors (ibrutinib, acalabrutinib, etc.).A systematic review of the scientific literature was performed using a step-by-step search process in electronic databases (PubMed, Web of Science, ScienceDirect, and Scopus). The following keywords were used in the database search: “CLL”, “BTK”, “ibrutinib”, “COVID-19”, “allergy”, “inflammation.” The search for studies was conducted from the time of the first BTK inhibitor drug (ibrutinib) appearance in 2009 until December 2022.The results of the study on the influence of BTK inhibitors on the functional state of B and T lymphocytes, neutrophils, and monocytes/macrophages are presented. The immunomodulatory effects of ibrutinib on adaptive and innate immune system cells, including CD4+ and CD8+T lymphocytes and NK cells, are described. Since BTK inhibitors alter the functional activity of phagocytic cells and the ratio of T cell populations, there is a suggestion about the possibility of using these drugs for the treatment of other nosological forms, not only B cell malignancies, which is currently being studied in clinical trials. Data on the use of BTK inhibitors to combat hyperacute inflammation and to suppress allergic reactions, including anaphylaxis, are summarized. In addition, the expediency of short-term use of BTK inhibitors to reduce the risk of side effects during oral immunotherapy and for desensitization to drugs is discussed.The presented data indicate that BTK inhibitors are promising drugs with immunomodulatory effects. However, BTK inhibitors need to increase selectivity to reduce off-target effects on other kinases.

Daily inhaled antigens induce cellular immune response in the airways. In case of allergens, allergic airway inflammation is usually represented by eosinophils, however, neutrophil infiltration is also observed during severe asthma. Animal models contribute to investigation of the mechanisms that involve the switching to eosinophil- or neutrophil-mediated inflammation. Data about the spatial location of eosinophils and neutrophils in the airways are necessary for both the understanding of allergic airway inflammation mechanisms and the drag potential estimation, however, not completely investigated. In the present study, we characterized the model of Aspergillus fumigatus extract-induced allergic airway inflammation that allowed investigating the early stage of inflammation development. The model adequacy was confirmed according to the blood and bronchoalveolar lavage eosinophilia. Using immunohistochemical staining of conducting airway as a whole-mount and confocal laser scanning microscopy, we estimated neutrophil and eosinophil spatial location: in the luminal side of the epithelium, in the airway wall or in the submucosal compartment close to the smooth muscle layer. An allergic airway response activation was detected upon significant elevation of blood eosinophil percentage compared to intact mice. Simultaneously, the number of eosinophils in the bronchoalveolar lavage was also significantly increased compared to the intact mice. At this time point, eosinophils predominated both in bronchoalveolar lavages and in conducting airway mucosa compared to neutrophils. Spatial location of conducting airway mucosal cell analysis demonstrated that eosinophils mostly located in the submucosal compartment, in a lesser extent in the airway wall, and a few eosinophils were detected in the luminal side of the epithelium. Neutrophils mainly infiltrated the luminal side of the epithelium, and a few neutrophils were detected in the submucosal compartment, while no neutrophils were detected in the airway wall. The data suggests that in response to the further allergen challenge, evidently eosinophils but not neutrophils will migrate through the airway wall to the airway lumen. Thus, eosinophils can be expected to damage airway epithelium in allergic airway inflammation development. Simultaneously, neutrophils located in close proximity to the smooth muscle layer together with eosinophils can contribute to bronchoconstriction induction.

Recurrent urinary tract infections (UTIs) are associated primarily with the ability of Escherichia coli to form biofilms. The interaction of neutrophils, factors of innate immunity, with microorganisms in biofilms is difficult compared to planktonic forms due to the lack of direct contact, as well as due to the antiphagocytic action of the extracellular matrix of biofilms. The purpose of this study was evaluation of neutrophils phagocytic and oxidative activity during interaction with biofilms of uropathogenic E. coli (UPEC) DL82 and R44. Peripheral blood neutrophils from healthy men were isolated using ficoll-urographin double gradient, incubated for 1 h with bacterial cells from biofilms or their supernatants, then leukocytes functional activity was evaluated. Phagocytic activity of neutrophils was determined by the degree of bioluminescence inhibition of bioluminescent strain E. coli K12 TG1 lux+ (pXen) upon their absorption by neutrophils. Production of extracellular reactive oxygen species (ROS) was analyzed by the intensity of luminol-dependent chemiluminescence in spontaneous and stimulated by E. coli K12 variants. Significance of differences was determined using Student’s t-test at p < 0.05. It was found that neutrophils interaction with UPEC biofilm cells or supernatants did not affect the phagocytic activity. E. coli DL82 supernatants reduce neutrophils spontaneous ROS production compared to control and biofilm cells. E. coli R44 supernatants with a low virulence potential did not affect ROS production, while biofilm cells stimulated it. When assessing stimulated ROS production, exposure to R44 strain supernatants did not cause a decrease in neutrophils activation in response to an external stimulus (E. coli K12 cells). Preliminary contact of neutrophils with E. coli R44 bacteria resulted in a high and prolonged level of ROS production compared to the control. Neutrophils interaction with DL82 cells resulted in a higher level of ROS compared to supernatants, however a subsequent rapid depletion of neutrophils oxidative potential was observed. Thus, cells and supernatants of UPEC biofilms can determine the activation of neutrophils.

Osteoarthritis is caused by a complex interplay of genetic, metabolic, immunological, inflammatory, biochemical, and biomechanical factors. In recent years, a fairly large number of studies have been devoted to the role of cellular factors of the immune system in the development of osteoarthritis. The aim of the study was to analyze scientifc publications devoted to the study of cellular factors in the pathogenesis of osteoarthritis and to assess their signifcance in the development of joint pathology. Material and methods. The search for publications by keywords was carried out in the PubMed, Google Scholar, eLibrary databases and specialized journals related to therapy, rheumatology, traumatology and immunology from 2000 to 2022. Results and discussion. Summarizing modern ideas about the role of cellular factors of the immune system in the pathogenesis of osteoarthritis, it is necessary to note the presence of synovial inflammation, a key role in the development of which is assigned to macrophages. At the same time, patients with osteoarthritis are characterized by the predominance of classically activated macrophages with a pronounced pro-inflammatory effect. In addition, T lymphocytes also play an important role in the pathogenesis of joint damage. Among them, a special role is given to T helper cells, cytotoxic T lymphocytes and memory T cells. An imbalance of cytokines and chemokines produced by subpopulations of T lymphocytes is the reason for triggering a number of mechanisms for the onset and progression of osteoarthritis. A signifcant role in the development and progression of osteoarthritis is also assigned to neutrophils, which contribute to the development of inflammation. Neutrophil-produced elastase enhances cartilage degradation, chondrocyte apoptosis, unbalanced subchondral bone remodeling, and osteophyte formation. Conclusions. Knowledge of the role of cellular immune factors in the pathogenesis of osteoarthritis and ways to implement their effects determines the prospects for the use of immunotropic agents. Also, it should be taken into account that the occurrence and progression of osteoarthritis is due to the simultaneous combination of the influence of a wide range of various components, including risk factors, traumatic joint injury, etc.

The studies were carried out in 24 sexually mature male guinea pigs. They were used as model of immediate-type allergic reaction (active cutaneous anaphylaxis). The animals were divided in 2 groups: control and experimental. The control group was housed in the mountain valley during the study, and the experimental group stayed for 45 days in high mountains (Anzob, 3375 m above sea level). On the day +30, all animals were sensitized with horse serum. On the 12th day of sensitization, blood was taken for analysis, and on the 15th day, an allergic reaction was provoked. We have revealed that, under high-altitude conditions, the severity of reaction was 1.5 times lower than in controls. Moreover, the animals kept in highlands exhibited lower contents of T and B lymphocytes, and IgE antibodies than in the control group. On the contrary, the numbers of phagocytically active neutrophils, as well as total effect of phagocytosis, proved to be higher in this group. This shift may be facilitated by hypoxia, since aerobic processes are known to prevail in the energy metabolism of lymphocytes, and anaerobic processes dominate in neutrophils. The allergic conditions are developed in three stages. Immunological stage is the first and main one, and allergic restructuring of the body immunity largely depends on it. What is the matter of reconstruction under the high-altitude conditions? We suggest, that, along with Haeckel–M ller biogenetic rule, there is also an immunogenetic law: “The activation sequence of events in the body’s defense system is a reproduction of phylogenesis, i.e., switching of subsequent link in defense system follows the evolutionary principles of complication and improvement. Namely, with respect to failing of previous link to completely eliminate the antigen in defense system”. One may conventionally consider that the first protective barrier against antigens is the most evolutionarily ancient structure, i.e., skin and mucous membrane; the second represents factors of nonspecific defense (phagocytosis, lysozyme, interferon, etc.); the third barrier is presented by cellular immunity (T effector cells, etc.), with humoral immunity serving as the fourth barrier. The fifth protective barrier provides evolutionarily late defense, i.e. allergic reaction, which is triggered by Ig E antibodies. On the mentioned basis, one may state that the decreased allergic reaction (the 5th barrier) in the animals from the experimental group was caused by adaptive changes in protective mechanisms of body. One may suggest that, under the high-altitude conditions an increase was observed in functional activity of phagocytes (2nd barrier), along with a decrease in activity of lymphocytes, i.e., cell populations responsible for the evolutionary later protective barriers.

Granulocyte-macrophage colony stimulating factor (GM-CSF) is a myelopoietic growth factor that exerts pleiotropic effect not only on the differentiation of immature progenitor cells into polymorphonuclear neutrophils, monocytes/macrophages and dendritic cells, but also controls the functioning of differentiated cells. GM-CSF is currently being investigated in clinical trials as an immunomodulator and adjuvant. However, a wide range of biological activities and, sometimes, paradoxical effects of this cytokine require more thorough studies of its action, in order to predict its efficacy under different conditions of immunotherapy. In this work, we have studied the effect of recombinant human GM-CSF on metabolic activity of mouse peritoneal exudate cells in primary cell cultures. Metabolic (redox) activity of the cells was assessed by their ability to reduce nitroblue tetrazolium (NBT) in the course of MF- and Fc-dependent phagocytosis triggered by addition of opsonized zymosan, or sheep erythrocytes to the culture medium. We have shown the dose-dependent stimulatory effect of GM-CSF on the oxidative metabolism of phagocytic peritoneal macrophages and neutrophils. Upon culturing the pepton-elicited cells at wide range of GM-CSF concentrations (5 to 40,000 ng/mL) for 2 and 24 hours, a more pronounced effect of the substance was observed for neutrophils. The GM-CSF preparation caused a significant increase (by 13-17%) in the redox activity of neutrophils induced by opsonized zymosan that persisted at a low dose range, and was retained after 24 hours. The stimulatory effect of GM-CSF on macrophages with NBT index increase by 16% was observed in the short-term cultures. In general, the elicited cells of both types showed a more pronounced response to lower concentrations of GM-CSF (5-125 ng/mL), and weaker effect at higher doses of the preparation. A similar dependence was found when studying the resident macrophages. Culturing of resident cells with GM-CSF at the doses of 5,000 to 40,000 ng/mL for 24 hours caused a significantly increased redox activity of the cells induced by zymosan, or sheep erythrocytes (by 33-52%). In both cases, the maximal response was detected at a dose of 5,000 ng/mL and decreased with increasing dose. The stimulatory effect of GM-CSF upon resident macrophages was more pronounced as compared to elicited cells, which was characterized by the prolonged period of cell activation (up to 24 hours of culture). The data obtained are of interest, in view of prospective usage of GM-CSF as a component of immunomodulatory and adjuvant therapy for various infectious diseases.

Introduction. One of the leading causes in the mortality pattern of cancer patients is accounted for by thrombotic complications. Recent studies have shown that neutrophil extracellular traps (NETs) are involved in the activation of coagulation, contribute to the initiation and progression of thrombosis. In addition, NET-related effect on tumor progression and metastasis has been actively studied.Aim: to evaluate NET-related procoagulant activity in gynecological cancer patients.Materials and Methods. From April 2020 to October 2022, a prospective controlled interventional non-randomized study was conducted with 120 women. The main group included 87 patients aged 32 to 72 years with malignant neoplasms of the female genital organs and mammary glands who were hospitalized for elective surgical treatment or chemotherapy: uterine body cancer (subgroup 1; n = 18), ovarian cancer (subgroup 2; n = 26), cervical cancer – adenocarcinoma of the cervical canal (subgroup 3; n = 13), breast cancer (subgroup 4; n = 30). The control group consisted of 33 healthy women aged 32 to 68 years. In all women, plasma concentrations of citrullinated histone H3 (citH3), myeloperoxidase antigen (MPO:Ag), D-dimer, and thrombin–antithrombin (TAT) complexes were evaluated.Results. The magnitude of NETosis in cancer patients, assessed by level of citH3 (2.5 ± 0.7; 1.9 ± 0.8; 2.5 ± 0.7; 0.7 ± 0.5 ng/ml in four subgroups, respectively) and MPO:Ag (29.5 ± 13.1; 12.8 ± 3.7; 22.8 ± 8.7; 6.6 ± 2.5 ng/ml in four subgroups, respectively) was significantly higher compared to women in the control group (0.3 ± 0.1 ng/ml; p = 0.0001 and 2.5 ± 0.2 ng/ml; p = 0.0001). In parallel with increased NETosis markers in accordance with the disease stage, there was an increase in the concentration of hemostasis activation markers – D-dimer (1.7 ± 0.6; 2.0 ± 0.7; 1.4 ± 0.5; 1.5 ± 0.7 µg/ml in four subgroups, respectively) and TAT complexes (729.8 ± 43.9; 794.1 ± 164.8; 636.2 ± 149.5; 699.6 ± 165.7 pg/ml in four subgroups, respectively) exceeding their level in the control group (respectively, 0.4 ± 0.1 μg/ml; p = 0.0001 and 362.3 ± 0.1 pg/ml; p = 0.0001). The maximum values of parameters occurred at later stages according to the Classification of Malignant Tumours (tumor, nodus, metastasis, TNM). A significant correlation between TAT level and the concentrations of citH3 (r = 0.586; р = 0.04) and MPO:Ag was revealed (r = 0.631; р = 0.04).Conclusion. Tumor tissue creates milieu that stimulates NETs release, which, in turn, not only contribute to the creating a procoagulant state, but also might act as one of the factors that ensure tumor progression and metastasis. The development of targeted therapies acting on NETs has a potential to affect hemostasis in cancer patients and reduce rate of tumor growth and metastasis.

Atrial fibrillation and ischemic heart disease are the key problems in cardiology. Despite of numerous clinical trials and researches underlying molecular biology remains uncertain. Atrial fibrillation and ischemic heart disease are often combined. During ischemic heart disease progression myocardial tissue structure are changing which lead to structural and electrophysiological remodeling and promote atrial fibrillation. It has been shown a crucial role of oxidative stress and chronic systemic inflammation in ischemic heart disease and atrial fibrillation. Myeloperoxidase (MPO) is one of marker of oxidative stress and inflammation that located in azurophilic granules of neutrophils and monocytes. There are a numerous articles showed a relation between MPO level and cardiovascular disease. MPO is a peroxidase enzyme that is important part of immune system. During disease MPO could facilitate chronic inflammation and local tissue damage through active oxygen forms. MPO releases after lysosome conjunction with phagosome. Oxygen reductase activity of MPO lead synthesis of hypochlorous acid that play role not only in organism protection from infection agents but in matrix transformation and fibrosis. It has been shown MPO can destabilize atherosclerotic plaque and modifies low- and high-density lipoproteins that promote atherosclerosis and ischemic heart diseaseу progression. This review summarizes current data about role of MPO in atrial fibrillation and ischemic heart disease pathogenesis.

Objective. Studying of mutual role of the main pathogenetic factors of abdominal sepsis – the inflammation origin and intraabdominal hypertension, from the one side, endotoxicosis and the abdominal compression syndrome (ACS), from the other side. Materials and methods. Into the investigation 196 patients with abdominal sepsis were included, in whom after the certain preliminary preparation the operative procedure was done. The levels of neutrophils, macrophages, interleukin–6, interleukin–10 in peritoneal exudate were studied as indices of the inflammation source for abdominal cavity. These indices were characteristic for systemic answer on general reaction, the degree of which have determined the abdominal sepsis course and result. Determination of intraabdominal pressure was applied for estimation the intraabdominal hypertension role in abdominal sepsis. Results. The ratio of the endotoxicosis and the ACS indices in the complications structure, not dependent on some differences due to the complication type present, was practically similar. Conclusion. The results of the investigation conducted, witnesses the presence of interrelationship of roles of endotoxicosis and the abdominal compression syndrome in pathogenesis of abdominal sepsis, their indices changes are characterized by mutual activity strengthening, accompanied by clinical manifestations of the organs dysfunction, determining the severity of course and result of pathological process in the organism.

Purpose. To investigate the cell composition of the tear in healthy women and endometriosis patients in order to find the potential diagnostic criteria of endometriosis. Material and methods. Tear samples obtained from the lower fornix of the conjunctiva using a glass capillary tube were microscopically examined in 100 females aged 18 to 48: 60 healthy women, who made up the control group, and 40 patients with endometriosis. Results. The share of women with endometriosis who were found to have erythrocytes was 32.5 %, which was significantly higher as compared with the control group, where it was 25.0%. 69.2 % of women with endometriosis showed haemolacria in the follicular phase of the menstrual cycle, which significantly exceeded the percentage of women with erythrocytes in the tear (30.8 %) who had them in the luteal phase. Epitheliocytes were found in the tear of 97.5 % of patients with endometriosis, versus 75.0 % of healthy women. In all 100 % of such patients, these cells were found in the follicular phase of the menstrual cycle, compared with 74.3 % of healthy women who showed these cells in the same phase of the cycle. The share of women with endometriosis showing the ++ quantity of epitheliocytes in the tear was 56.4 %, which was significantly higher than the 20 % of the control group subject with the same level of epitheliocytes. It is to be noted that group accumulations of epitheliocytes were found in 15 % of endometriotic patients, which was significantly higher than in healthy women, of whom only 8.3 % had such accumulations. 90 % of endometriosis group cases revealed glandular cuboidal epithelial cells in the tear, which morphologically resembling endometrial cells. Conclusion. The revealed properties of the cytological composition of the tear of patients with endometriosis reflected particular links in its etiopathogenesis, which suggests that the study of tear parameters can help predict the incidence of the "endometrioid disease" and develop pathogenetically oriented treatment methods.

The results of recent studies demonstrating the presence of pro-inflammatory mediators in osteoarthritis (OA), as well as known similarity of histological signs of inflammation in rheumatoid arthritis (RA) and OA, suggest an important role of NETosis in immune inflammation in OA patients. Our objective was to assess ability of blood neutrophils from ОA patients to generate NETosis spontaneously and after in vitro induction, and impact of reactive synovitis on the dynamics of NETosis indexes. Thirty-one patients with verified OA and 30 healthy volunteers were included into the study. Circulating neutrophils were isolated with one-step density gradient centrifugation using double layers of Iohexol gradient. Subpopulational profile of isolated neutrophil fractions, their viability, and nonspecific activation were evaluated microscopically using Trypan Blue exclusion test, as well as nitro-blue tetrazolium test. NETs were induced by phorbol-12-myristate13-acetate (PMA). Spontaneous and induced formation of NETs was assessed using fluorescence microscopy. The ОA patients were in clinical remission at the time of inclusion in the study. In 23 OA patients, an exacerbation was diagnosed during the study. The neutrophil fractions showed high purity and a high content of viable nonactivated cells. These parameters were comparable in the study groups. Mean percentage of spontaneous NETs in OA patients in remission was significantly increased comparing to healthy controls. Usage of PMA, as inducing agent was accompanied by a significant increase in ability of neutrophils to form NETs. Transition of OA to exacerbation was characterized by further significant increase in spontaneous and PMA-induced NETs. Spontaneous and induced NETs in OA patients at acute stage of the disease are significantly higher than in OA patients in remission state. The growth rate of spontaneous NET formation is 3.74 times higher than the induced NET formation in OA patients upon exacerbation. Statistically significant increase in the ability of peripheral neutrophils to spontaneous and induced formation of extracellular traps was found, depending on the stage of osteoarthritis. The data obtained suggest an opportunity for participation of circulating neutrophils via NETosis in pathogenesis of immune inflammation in OA.

Background. Due to their very small size, nanomaterials, in particular cucurbiturils, have unique physical and chemical properties that find their application in medicine. However, the toxicity of cucurbiturils is not fully understood; in particular, we are interested in the immunological safety of their use. One of the mechanisms of nanotoxicity is the formation of reactive oxygen species (ROS) by macrophages and neutrophils. Hyperproduction of ROS can lead to oxidative stress and further damage to cell DNA with loss of physiological function and development of pathology. The aim. Evaluation of the effect of cucurbit[n]urils (n = 6, 7, 8) on the production of reactive oxygen species by mice macrophages and neutrophils. Materials and methods. F1 hybrid mice (CBAxC57Bl/6) aged 2 months (n = 11) were used in the work. Evaluation of superoxide radical production by peritoneal mouse neutrophils and macrophages was carried out by spectrophotometric method for determining the reduction of p-nitroblue tetrazolium (NBT) to formazan. Results. It was shown that CB[6] and CB[7] at concentrations of 0.5 and 0.3 mM do not have an inhibitory effect on ROS synthesis, but, on the contrary, significantly increase ROS production by macrophages. In addition, CB[6] 0.3 mM increases the level of ROS in neutrophils. Conclusion. Cucurbiturils can lead to an increase in the production of ROS in immunocompetent cells, depending on the concentration used (0.3 mM and higher).

Relevance. The inflammatory process and endogenous intoxication of the patient’s body has a negative impact on the course of many malignant neoplasms, including salivary gland cancer.The objective of the study was to determine the influence of some factors of systemic inflammation and endogenous intoxication in salivary gland cancer.Methods and materials. A prospective study included the data of 59 patients with salivary gland cancer. The influence of peripheral blood parameters and relative indicators characterizing systemic inflammation on overall and disease-free survival was assessed.Results. As a result of the multivariate analysis, it was revealed that an increase in the level of the relative number of peripheral blood neutrophils by more than 60.08 % increases the risk of death in patients from salivary gland cancer by 3.90 times (p=0.0456; HR 3.90: 95 % CI 1.03–14.79). The level of the absolute number of peripheral blood lymphocytes, not exceeding 1.49x109 /l, increases the risk of disease progression by 8.72 times (p=0.0002, R 8.72: 95 % CI 2.78–27.28).Conclusion. Individual factors of systemic inflammation and endogenous intoxication, it is advisable to evaluate at the stage of planning the primary treatment of patients with salivary gland cancer, in order to determine the prognosis of the disease and optimize the choice of tactics for the primary treatment of patients.

Rheumatoid arthritis (RA) ranks first among chronic joint diseases. The disease often affects people at their working age, being accompanied by significant decrease in the life quality of patients and their early disability. Rheumatoid arthritis is an immunoinflammatory rheumatic disease. Therefore, the immune system provides evolving focus of primary damage, its persistence and periodic exacerbation. Elucidation of intercellular relationships mediated by cytokines at various stages of the chronic inflammatory process is required in order to develop immunotherapeutic approaches, aimed for both recovery from exacerbations and maintenance of remission state. Purpose of our study was to evaluate cellular composition and cytokine profile of synovial fluid in the patients with rheumatoid arthritis at acute phase and in remission state.We have studied the samples of synovial fluid taken in 60 patients with rheumatoid arthritis, with 30 subjects being at acute stage of the disease, and 30 patients in remission. Cellular composition and cytokine profile were assessed in the clinical samples. There were 21 women and 9 men at the acute stage (57.0±15.4 years old), with the disease duration of 8.55±6.9 years. The average age of 19 women and 11 men examined in remission state was 53.5±10.9 years, with comparable duration of illness (6.9±5.8 years). The leukocyte phenotyping was performed with a CytoFLEX flow cytometer (Beckman Coulter, USA). The cytokine contents were measured by enzyme immunoassay using a standard set of reagents from the “Proteinovy Contour” LLC (Russia). The results were registered by a Multiscan photometer (Labsystems, Finland).During the disease exacerbation, the leukocyte contents in synovial fluid increased 2.4-fold, as compared to the remission values. The cellular infiltrate was represented by neutrophils, whereas the contents of lymphocytes and monocytes did not change. Increased migration of neutrophils was accompanied by an 8-fold increase in TNFα levels, compared with remission state, and IL-1β levels were increased by 6.3 times. The absolute number of CD3+T lymphocytes, CD16+CD56+B cells, and CD3-CD19+NK during exacerbation was similar to the remission levels. However, the number of T cell subpopulations was changed, i.e., the number of CD4+ lymphocytes was decreased, and CD8+ cell counts were increased, like as numbers of Treg lymphocytes and NKT cells which showed a significant increase. A 4.3-fold increase in the IL-4 concentration during the RA exacerbation suggested the predominance of Th2 immune response. During remission, the concentrations of IL-6 and IFNγ in synovial fluid were increased, respectively, by 1.5 times and by 2.5 times, which is typical for activated Th1 response.

The aim of the present study was to investigate the possibility of using immunophenotyping of circulating lymphocytes, neutrophils and monocytes to diagnose prognostically unfavorable carotid atherosclerosis using machine learning algorithms. Material and methods. A sample of patients aged 40 to 64 years, who underwent duplex scanning of the carotid arteries and the lower limb arteries, served as a source of patients for analysis. Phenotyping and differentiation of subpopulations of lymphocytes, neutrophils, and monocytes were performed by flow cytometry using the “Navios 6/2” device (Beckman Coulter, USA) device. Data were statistically processed using software package SPSS 23 and our own software programs created using main libraries of Python 3.8 programming language (Scikit-learn, Pandas, Numpy, Scipy) and IDE Spyder. Results. Seventy-eight patients, 39 (50.0 %) males and 39 (50.0 %) females, median age 50.0 years, were included in the study. Age over 56 (р = 0,001), elevated low density lipoprotein (LDL) cholesterol (р < 0.001) and uric acid (р = 0.001), as well as immunosuppressive neutrophils level (р = 0.005) were statistically significantly associated with the development of carotid plaque. At the same time, decreased cell populations of proangiogenic neutrophils (р = 0.009), classical monocytes expressing CD36 (р = 0.019), nonclassical monocytes (р = 0.021), intermediate monocytes expressing TLR4 (р = 0.033), and classical monocytes expressing TLR2 (р = 0.044) were statistically significantly associated with an increased chance of carotid plaque. Taking into account the selected parameters, two prognostic models were created. The first model included basic clinical and laboratory parameters (age, LDL cholesterol and uric acid), and the second model included all selected parameters as well as immunological parameters. Inclusion of the identified immunological predictors in the model resulted in a significant increase in all standard quality metrics of the binary classification. Model accuracy increased by 13 % (р = 0.014), sensitivity by 20 % (р = 0.003), specificity by 6 % (р = 0.046), predictive value of a positive result by 9 % (р = 0.037), predictive value of a negative result by 16 % (р = 0.011). According to the ROC analysis, without the inclusion of immunological predictors in the model, the area under the ROC curve (AUC) was 0.765 [0.682; 0.848], the inclusion of immunological predictors resulted in a statistically significant increase in AUC (0.906 [0.854; 0.958], р = 0.041). Conclusions. In patients 40–64 years old without established atherosclerotic cardiovascular disease, inclusion of immunological markers derived from leukocyte immunophenotyping in the model allowed increasing its diagnostic efficacy with regard to the detection of prognostically unfavorable carotid atherosclerosis. Subpopulations of monocytes expressing TLR2, TLR4, and CD36, as well as immunosuppressive and proangiogenic neutrophils, demonstrated diagnostic value.

Behcet's disease (BD) is a systemic vasculitis of unknown etiology, characterized by damage of vessels of any type and caliber, manifested by recurrent ulcerative process in the oral cavity and genital organs, eye damage, joints, gastrointestinal tract, central nervous system and other organs involvement. The pathogenesis of the disease is complex. The contribution of both innate and adaptive immune responses is noted. A feature of BD is hyperactivation of neutrophils and neutrophilic infiltration of affected tissues.The review presents data from studies related to the assessment of the main functions of neutrophils in this disease.

The incidence of ulcerative colitis has been increasing in recent years, and its manifestation at a young age has become a trend that is prognostically unfavorable. The clinical picture of ulcerative colitis is often vague, which leads to an initially erroneous diagnosis. One of the main problems is to assess the effectiveness of treatment and the risk of recurrence of ulcerative colitis, which requires invasive intervention. The aim of the study was to analyze the data of modern scientific literature on noninvasive biomarkers of ulcerative colitis. The data of foreign and domestic articles on the research topic published in Pubmed and eLibrary over the past 5-10 years are analyzed. Biomarkers of neutrophil origin are a promising direction in the primary diagnosis and assessment of ulcerative colitis activity.

Complexity and multifactorial nature of potential pathogenic consequences of SARS-CoV-2 infection in human body, discovery of new virus-induced mechanisms triggering a cascade of pathological responses in the cells of host organism leading to development of multiple organ failure elicited increasing interest in morpho-functional state of blood cells in reconvalescent persons after COVID-19 infection. The aim of the present work is to characterize morphofunctional pattern of blood cells at different periods of recovery, depending on the severity of COVID-19. We examined 55 convalescents after bearing COVID-19 infection: Group I included the convalescents 30 days after the disease (n = 39); Group II consisted of the persons 60 days after recovery (n = 16); Group III included clinically healthy volunteers with no history of clinical SARS-CoV-2 infection (n = 11). The cells were examined by means of Olympus CX41 microscope (Olympus, Japan), and VZ-C31S digital videocamera (VideoZavr, Russia) using the VideoZavr software (version 1.5). Assessment of neutrophil populations in the whole blood samples was performed with BD Accuri C6 Plus flow cytometer (USA) with automatic differentiation of cells between lymphocytes and monocytes, according to the degree of granularity. Cytokine production was determined using commercial kits for detection of IFNγ, TNFα, IL-4, IL-8, IL-10 (JSC Vector-Best, Russia), IL-17A (eBioscience, Austria) was assayed with automatic enzyme immunoassay analyzer “LAZURIT” (Dynex Technologies, USA). Among the convalescents who suffered the moderate-degree COVID-19 (45.5% and 50% of cases, respectively) on days +30 and +60 after clinical recovery, a significantly increased ratio of morphologically altered forms of erythrocytes (echinocytes, ovalocytes, dacryocytes, codocytes) was noted as compared with group III (p = 0.00001 and p = 0.001, respectively). Regardless of clinical severity of the disease; a mean of 40.6% convalescents from groups I and II had moderate disturbances in the neutrophil morphology (cytoplasmic vacuolization, chromatin decondensation at the pre-netosis stage, transformation of cells by the netosis type), and, in 27.4% of cases, the areas of neutrophilplatelet aggregation were seen. In blood supernates from recovered patients, we have revealed a significantly decreased content of IFNγ (P = 0.02), TNFα (p = 0.03), IL-10 (p = 0.04) and IL-17A (p = 0.02). The revealed morphological and functional changes in blood cells in the persons who underwent COVID-19 infection suggest long-term maintenance of toxic damage to erythrocytes, neutrophils and lymphocytes over the recovery period. The effects of the detected morphological and functional disorders of blood cells following COVID-19 recovery leading to increase in blood viscosity and microcirculation, formation of neutrophil-platelet aggregates, may cause higher risks of thrombotic complications at the long-range period as well as decreased levels of regulatory cytokines, thus confirming slow recovery of the lymphocyte populations (Th1, Th2, Th17) of the immune system.

Chronic rhinosinusitis (CRS) affects 5-12% of the world’s adult population. Chronic rhinosinusitis with nasal polyps (CRSwNP) accounts for 25-30% of all cases of CRS. CRSwNP-associated inflammatory process in nasal mucosa and paranasal sinuses depends on the characteristics of local immunity, including expression of a number of cytokines. The aim of this work was to investigate the parameters of local immunity in various clinical forms of CRSwNP. In this work, the concentrations of pro-inflammatory cytokines, i.e., interleukin-1β (IL-1β) and IL-8, antimicrobial function of neutrophils from the nasal cavity was evaluated, along with histological and immunohistochemical studies of polyposis tissue. The study included 4 groups of patients: a control group of practically healthy individuals, patients with CRSwNP, clinical cases with chronic purulent rhinosinusitis and nasal polyps (CPRSwNP), and patients with CRSwNP complicated by bronchial asthma (CRSwNP + BA), including the cases with asthmatic triad (CRSwNP + intolerance to NSAIDs + BA). The patients were classified on the basis of their clinical characteristics and severity of the course of the disease. Interleukin-1β (IL-1β) and IL-8 concentrations in nasal secretions were determined by enzyme-linked immunosorbent assay (ELISA). To assess functional activity of neutrophils, a lysosomal cationic test was used on the smears from mucous surface of inferior turbinate. Histological examination of the polypous tissue biopsies was performed in slices stained with Carazzi’s hematoxylin and eosin. IL-1β and IL-8 location in the polypous tissue were detected by indirect immunohistochemistry. In all groups of the patients, IL-1β and IL-8 concentrations exceeded those in the control group. The levels of IL-1β in the groups with CPRSwNP, CRSwNP + BA were significantly increased as compared with the CRSwNP group. IL-8 concentrations in the CRSwNP and CPRSwNP groups were significantly higher than in the CRSwNP + BA group. When analyzing antimicrobial function of neutrophils, the decreased average values of cytochemical coefficient were shown in CPRSwNP and CRSwNP + BA groups, compared with the control group and CRSwNP. In all clinical forms of CPMS, complex histopathological changes were observed, including leukocyte infiltration, fibrosis, edema, and collagen depositions. In addition, the integrity of epithelial layer was found to be damaged in polyposis, epithelial metaplasia is detected as well as increased mucus production. These disorders lead to a decrease in muco-ciliary clearance in nasal cavity. The most significant pathomorphological changes occur in CRSwNP + BA, especially in cases of asthmatic triad. According to immunohistochemical data, in various forms of CRSwNP, IL-1β- and IL-8-positive leukocytes, predominantly macrophages, are detected in the polypous tissue both subepithelially and in the connective tissue stroma of the polyps. Changed concentrations of pro-inflammatory cytokines in nasal secretion of the patients, altered antimicrobial activity of mucosal neutrophils, and characteristic pathomorphological disorders in polypous tissue of patients with CRSwNP are associated with severity of inflammatory process and clinical course of the disease. The results obtained are essential to understanding the mechanisms of pathogenesis in various subgroups of CRSwNP, assessing severity of the disease and efficiency of the treatment applied.

Background: C-reactive protein (CRP) is a key laboratory biomarker for anti-inflammatory treatment initiation. Unfortunately, biochemical blood analyzers are not always easily accessible in medical institutions located far from major regional health facilities. Aim: To develop an approach for inflammatory status estimation based on blood test results in patients with COVID-19 in cases with limited laboratory equipment availability. Methods: The present retrospective study included 423 patients (male 54.6%; female 45.4%; mean age 59.1 years) receiving hospital treatment due to COVID-19 in Medical Research and Educational Center of Lomonosov Moscow State University from April 21st to June 13th 2020. All patients donated blood for full biochemistry and hematology testing and underwent chest computer tomography (CT). Results: CRP levels (60 mg/L) qualitative estimation model was developed based on hematologic test results. It included erythrocyte sedimentation rate and neutrophils to lymphocytes ratio. According to the results of Receiver Operating Characteristic (ROC) analysis present model was characterized by sensitivity of 70.2%, specificity of 74.6%, and area under the ROC-curve of 0.781. Comparison of key clinical parameters reflecting COVID-19 severity, such as length of hospitalization, lung damage at CT (hospital admission and discharge), revealed statistically significant difference between groups with routinely measured CRP levels 60 mg/L and 60 mg/L for all the above-mentioned parameters (p 0.05). These differences remained significant when measured CRP levels were substituted with estimated CRP values, indicating interchangeability of these approaches to CRP levels determination, regarding clinically important parameters. Conclusions: Presented model for inflammatory status estimation based on hematologic test results might be used to overcome clinical challenges in cases with limited laboratory equipment availability.

The last fifteen years have been marked by rapid progress in the study of neutrophils. The discovery of transcriptional plasticity of neutrophils, their phenotypic and functional heterogeneity contributed to launching active interdisciplinary studies on the role of neutrophils in various chronic inflammatory diseases. Increased systemic circulation of immunosuppressive neutrophils can be observed not only in sepsis, but also in chronic systemic inflammation, which, along with disorders of lipid metabolism, is the major mechanism of atherosclerosis development and progression. Monocytes, dendritic cells, Tlymphocytes and neutrophils are key participants and modulators of inflammation in atherosclerosis. Potential significance of immunosuppressive neutrophils in atherogenesis and regulation of inflammatory response in atherosclerosis has not been currently established. However, taking into account their possible effects upon T lymphocytes and innate immunity cells, the study of immunosuppressive neutrophils seems promising in the context of atherosclerosis and atherosclerotic cardiovascular diseases. The purpose of this study was to evaluate relationship between the numbers of circulating immunosuppressive neutrophils and subpopulations of T cells and monocytes in the patients with subclinical atherosclerosis. The study enrolled patients aged 40-64 years with subclinical atherosclerosis of peripheral arteries. Subpopulations of neutrophils, lymphocytes and monocytes were phenotyped by flow cytometry using “Navios 6/2” (Beckman Coulter). 133 patients, 65 (48.8%) males and 68 (51.2%) females were included into the study. Correlation analysis showed that increased number of circulating CD16hiCD11bloCD62Lbr neutrophils was associated with increased number of regulatory T lymphocytes. The patients with subclinical atherosclerosis and absolute numbers of circulating immunosuppressive neutrophils within the first quartile (

Currently, there are only scarce data on dynamics of biologically active substances in the lesions associated with atopic dermatitis. Persistence of microorganisms in atopic dermatitis is high on the skin surface. However, pathophysiological significance of ENA-78/CXCL5 for development of atopic dermatitis was not studied so far. The ENA-78/CXCL5 is known to be produced by endotheliocytes, keratinocytes, eosinophils, fibroblasts to activate neutrophil migration, especially under the influence of LPS-containing microorganisms. The aim of this study was to evaluate the dynamics of ENA-78/CXCL5 chemokine levels in blood serum and skin exudates in the patients with atopic dermatitis, as well as to determine pathophysiological role of the chemokine in pathogenesis of dermatosis. 80 patients with limited and widespread forms of atopic dermatitis and 15 volunteers were under observation. The dynamics of ENA-78/CXCL5 levels was studied in blood sera and skin exudates. Blood samples for the study were drawn at the time periods of exacerbation and remission. Skin exudates were taken from the patients during the exacerbation period using disposable insulin syringes and 20-G disposable needles. In healthy volunteers, the skin exudate was obtained by the “skin window” technique as described by V.V. Klimov and coauthors “A method for assessing minimal inflammatory activity of skin in atopic dermatitis in remission”. The cell analysis was conducted by flow cytofluorimetry using the LEGEND plex TM Human Proinflammatory Chemokine Panel (USA) according to the manufacturer’s protocol. Serum concentrations of chemokine ENA-78/CXCL5 in adolescents with atopic dermatitis, exceeded the range for healthy volunteers. During remission of dermatitis, the chemokine level did not reach the indices in the control group. In adults, the ENA-78/CXCL5 concentration, both at the onset of symptoms and upon their resolution, was below the control levels. Maximal concentrations of ENA-78/CXCL5 chemokine were detected in the skin exudates. As based on our data on the dynamics of ENA-78/CXCL5 chemokine levels, it could be assumed that this substance may represent a sufficient link in pathogenesis of atopic dermatitis, by causing migration of neutrophils and monocytes to the affected area. The ENA-78/CXCL5 chemokine may be a marker of microbial pathogenesis and cellular damage in atopic dermatitis.

Priming and activation mediated by cytokines cause transient reactions of actin polymerization in neutrophils (Nph), expansion and softening of cells, changes in receptor status, phagocytic ability, and generation of extracellular traps (NET), thus ultimately determining pro- or antitumor phenotype of Nph. To assess the effects of pro-inflammatory cytokines on membrane rigidity and morphofunctional state of neutrophils in benign tumors and ovarian cancer, the levels of circulating cytokines (IL-2, IL-18, MCP-1, TIMP-1), expression of adhesion markers (CD11b), degranulation (CD63), as well as FcãRIIIb receptors promoting phagocytosis (CD16). Ability of neutrophils to phagocytosis and the formation of NET was determined. Statistical evaluation of the data obtained was carried out using Statistica 13.0, Jamovi 1.6.5.0 software. An increase in membrane rigidity was found in benign and malignant ovarian tumors. In benign ovarian tumors, phagocytic activity and expression of CD11b were enhanced. In ovarian cancer, the number of CD11b+Nph and CD63+Nph were increased. Meaanwhile, at initial cancer stage, the ability to form NET predominates, and phagocytic activity increases with advancing ovarian cancer. Serum MCP-1 levels are elevated in benign tumors and at all stages of ovarian cancer. IL-2 is elevated at early stage and in advanced ovarian cancer. The level of IL-18 and TIMP-1 in the serum of patients with benign ovarian tumor did not differ significantly from the norm. Using multiple regression approach, the dependence of neutrophil membrane rigidity on the levels of circulating IL-2, TIMP-1, MCP-1, IL-18 was revealed in benign ovarian tumors, and a direct correlation was found between the neutrophil membrane rigidity and CD11b expression. Only IL-2 was associated with neutrophil membrane rigidity in ovarian cancer. At the same time, the rigidity of the neutrophil membrane directly correlated with CD16, CD63, expression like as with phagocytic index and inversely correlates with the number of traps. A combination of IL-2, MCP-1 and membrane rigidity of circulating neutrophils (based on multivariate analysis) could be used for differential diagnosis of ovarian cancer. Thus, in a benign ovarian tumor, circulating proinflammatory cytokines are associated with increased rigidity of neutrophil membrane and increase in their adhesion capacity. In ovarian cancer, only IL-2 is associated with altered rigidity of circulating neutrophils. Increase of the latter index is accompanied by elevated phagocytic activity and decreased ability to form NET.

Microflora of the oral cavity forms a biofilm that induces response of immune system at the mucous membranes. Transition to periodontal lesion is provided by certain classes of resident mucosal immune cells and inflammatory/immune cells migrating to the periodont. In periodontal diseases, Th1, Th2, Th17, Treg are detected. T regulatory cells (Tregs) are proven to comprise the main anti-inflammatory cell population. Th17 cells and Treg cells play an important role in osteoclast differentiation. IL-17 secreted by Th17 cells affects osteoclastogenesis and may induce macrophages to enhance the local inflammatory response. In this regard, the aim of our work was to identify the local immune cells in oral cavity which are associated with severity of chronic generalized periodontitis. The oral cavity cells from 58 persons aged 38-65 years of both sexes in their mature age with a diagnosis of «chronic periodontitis» were examined by means of flow cytofluorometry. When determining levels of CD64+CD16+CD14- neutrophils in the patients with periodontitis of different severity, a statistically significant increase of this cell population was revealed upon development of this disease. In mild cases of periodontitis, a significant increase of relative CD64+CD16+CD14- neutrophil contents was revealed (Me = 36.16%, p < 0.05) compared to the control group (Me = 7.7%, Q0.25 = 2.4%, Q0.75 = 12%). When assessing relative numbers of CD14+ monocytes in periodontitis of various severity, we revealed a significant increase in the number of these cells in severe cases. When studying levels of regulatory T lymphocytes (CD4+CD25+CD127low) in periodontitis of different severity, we revealed significantly decreased amounts of this cell population during development of the disease. In mild cases of periodontitis, a decreased level of CD4+CD25+CD127low cells (p < 0.05, Me = 1356 cells/ml) was revealed, as compared with control group (Me = 10666 cells/ml). Although the concentration of CD4+CD25+CD127low (Me = 4709 cells/ml) in the patients with moderate periodontitis was higher than the values in milder cases, the range of the main values was comparable and lower, than in control group. In severe periodontitis, a significantly decreased concentration of regulatory T lymphocytes was revealed (Me = 2637 cells/ml). These results confirm the anti-inflammatory regulatory function of Tregs. Understanding the osteo-immune mechanisms of bone remodeling control will help to understand the pathophysiology of accelerated bone loss observed in severe chronic periodontitis.

Aim. To study the activity of neutrophilic granulocytes in patients with opisthorchiasis, depending on the severity of liver fibrosis. Materials and methods. A total of 74 patients with chronic opisthorchiasis (39 men and 35 women, average age 42.3 years) and 32 practically healthy patients (17 men and 15 women, average age 41.5 years) aged 24 to 60 years were examined. Diagnosis of opisthorchiasis was carried out by two methods: coprooscopy and identification of eggs or bodies of adult parasites in duodenal content. Liver fibrosis was determined by the method of elastometry according to the METAVIR scale in all 74 patients with opisthorchiasis. The study of the functional activity of neutrophils in the blood was performed to all 74 patients with opisthorchiasis and 32 healthy individuals from the control group by chemiluminescent analysis with measurement of the reactive oxygen species (ROS) production intensity in a spontaneous and zymosan-induced reaction in lucigenin and luminol-dependent processes. Results. Liver fibrosis F2 by METAVIR was registered in 20.3% of the examined individuals, liver fibrosis F3F4 by METAVIR was detected in 17.6% of patients with opisthorchiasis. In patients with opisthorchiasis with liver fibrosis F3F4 by METAVIR, a significant decrease in the functional activity of neutrophilic granulocytes was registered in comparison with individuals with liver fibrosis F0F1 by METAVIR, as evidenced by a significant decrease in the maximum intensity of ROS production (Imax) and the area under the curve (S) chemiluminescence in lucigenin and luminol-dependent processes both in the spontaneous and zymosan-induced reaction. Conclusion. These results provide new information to explain the mechanisms of liver fibrosis in patients opisthorchiasis and create opportunities for the development of diagnostics and preventive technologies.

Tuberculosis (TB) continues to be an important and unresolved medical problem. About a quarter of mankind is infected with Mycobacterium tuberculosis, and about 5–10% of these people eventually develop TB. Macrophages and CD4+ T cells are considered the key cells providing defense against TB infection. The role of neutrophils in TB is less well defined. Neutrophils are short-lived granulocytes among first migrate into the infectious lung tissue and phagocy tose mycobacteria. On the one hand, there is evidence for protective role of neutrophils in TB released via anti-microbial peptides inhibiting mycobacterial growth, up-regulation of CD4+ T-cell activation, and dendritic cell migration in the lymph nodes. On the other hand, infection of genetically TB susceptible animals leads to an overwhelming lung neutrophil inflammation, development of necrotic granulomata, and a rapid death. Neutrophils act directly or indirectly on mycobacteria by different oxidative or other reactions including neutrophil extracellular traps (NETs) formation. Phagocytosis of mycobacteria by neutrophils is accompanied by the production of pro-inflammatory factors, thus making neutrophils active participants of inflammation in all stages of the infectious process. Finally, neutrophils die by apoptosis or necrosis. Necrosis of neutrophils, which is activated by reactive oxygen species, also prolongs the inflammation. In this way, there is strong evidence that neutrophils are the cells involved in the transition of infection to the terminal stage, participating in lung tissue destruction. Although neutrophils evolutionary developed many ways to resist pathogens, it is likely, that neutrophils do not possess sufficient anti-mycobactericidal capacities due to the development of many adaptations allowing mycobacteria to survive inside the neutrophils. Neutrophils effectively phagocytose but poorly kill mycobacteria, thus hiding bacilli from more efficient killers, macrophages, and playing the role of the “Trojan Horse”. In this review, we summarize the data on the involvement of neutrophils in TB inflammation. We discuss their ambiguous role in pathogenesis which depends upon mycobacterial virulence, host genetics, dynamics of migration to inflammatory foci, and persistence during initial and chronic stages of the infectious process.

Introduction. This article is devoted to the discussion of a life-threatening condition - basilar artery thrombosis (BAT). The article presents the results of a comparative analysis of two groups of patients with angiographically confirmed BAT: with and without reperfusion therapy.Relevance. Basilar artery thrombosis is a life-threatening condition in which mortality can reach 95%, and severe neurological deficits and dependence in everyday life on others (modified Ran-kin Scale (mRS), score 4–5) are observed in 65% of surviving patients. Ischemic stroke (IS) due to BAT is diagnosed in 1–4% of patients with acute cerebrovascular event (ACVE). Currently, reperfusion therapy (RT) in the form of systemic thrombolytic therapy (sTLT) with rt-PA and/or thrombectomy (TE) is the only effective and safe method for treating patients with IS. In randomized trials, it was found that RT, whether it is sTLT, TE or a combination of these techniques, despite the best degree of functional recovery in patients by day 90 from the onset of the disease, does not lead to a statistically significant decrease in mortality. Nevertheless, the results of 10 prospective studies of the use of endovascular treatment in patients with IS in the posterior circulation system indicate that this type of therapy leads to a decrease in 90-day mortality from 95% to 16–47% in case of successful recanalization (mTICI (modified Thrombolysis in Cerebral Infarction), 2–b–3).Aim of study. To assess the safety and efficacy of TE in patients with BAT.Material and methods. This study included 15 patients with IS, admitted within the 6-hour therapeutic window, and confirmed BAT by CT and/or MRangiography. The baseline level of wakefulness was assessed using the Glasgow Coma Scale (GCS), and the severity of neurological deficit was assessed by the National Institutes of Health Stroke Scale (NIHSS).Results. Basilar artery thrombectomy was performed in 7 patients (endovascular treatment group), 8 patients received no endovascular treatment (standard therapy group). Good clinical outcomes by day 90 from the onset of the disease (mRS, score 0–2) were observed in 57.1% of patients in the endovascular treatment group and in 12.5% of patients in the standard therapy group. However, these differences were not statistically significant (p>0.05). There were also no differences between the two groups in functional outcomes and Rivermead mobility index (RMI) by day 90 from the onset of the disease (Barthel Index of Activities of Daily Living (BI) score 97±5.0 and 100, RMI score 14.0±0.0 and 15 in the endovascular treatment group and standard therapy group, respectively). Mortality by day 90 in the endovascular treatment group was 42.9% (3 patients), in the standard therapy group — 87.5% (7 patients). In the subgroup of patients with GCS score at admission no more than score 8, there was a statistically significant strong two-way negative correlation between TE performance and mortality (r=-1.0; p=0.000), as well as a positive correlation between TE and a Rankine score by day 90 (r=0.956; p=0.003). Mortality was statistically significantly lower in the endovascular treatment group compared to the standard therapy group (0% versus 100%, respectively, p=0.025). Clinical outcomes of the disease by day 90 were statistically significantly better in the endovascular treatment group: mRS, score 0–2 in 100% of cases versus 0% in the standard treatment group (p=0.028).Conclusion. Currently, the only effective treatment for basilar artery thrombosis is thrombectomy. The possibility of performing thromboectomy in these cases should be considered in all patients, regardless of the severity of the stroke and the decrease in the level of wakefulness, since endovascular treatment in this case is a life-saving procedure. The symptom of a hyperdense basilar artery can be used as a diagnostic tool for suspected basilar artery thrombosis, but should not rule out angiography. In our series of observations, the mortality rate in patients who underwent thrombectomy was 42.9%. Good functional outcomes (mRS, score 0–2) and the absence mobility limitation (RMI, score 14) were observed in 57.1% of patients by day 90 after thrombectomy.

Background. The syndrome of systemic inflammatory response, which underlies the damaging effect of factors of infectious and non-infectious genesis, may cause multiple organ failure. The degree of its severity is determined, among other things, by the activation of neutrophils. The paper highlights new mechanisms of the anti-inflammatory action of the inhalation anesthetic xenon, mediated by a decrease in the ability of neutrophils to pro-inflammatory response.Aim of study. To evaluate the effect of xenon on the activation of human neutrophils under ex vivo conditions.Material AND methods. We studied the effect of xenon inhalation on reduction of the ability of neutrophils to be activated proinflammatory by reduced expression of adhesion molecules CD11b and CD66b on the surface of neutrophils and on the phosphorylation of proinflammatory kinases: ERK 1/2 and kinase — p38 in neutrophils of healthy volunteers.Results. The use of xenon at a dose of 30 vol. % within 60 minutes in healthy volunteers statistically significantly reduces the ability of neutrophils to proinflammatory activation. The addition of lipopolysaccharide (LPS) to the incubation medium of neutrophils causes their pronounced activation, statistically significantly increasing the phosphorylation of key proinflammatory neutrophil kinases ERK1/2 and kinase p38. Inhalation of xenon in volunteers (30% within 60 minutes) has a pronounced anti-inflammatory effect on LPS-stimulated neutrophils, decreasing their activation by inhibiting pro-inflammatory kinase ERK1/2 and pro-inflammatory MAP kinase p38.Conclusion. The actual study, performed on isolated neutrophils from volunteers who underwent xenon inhalation, revealed the anti-inflammatory properties of the inert gas xenon, which, in our opinion, may have a direct relationship to the identification of the mechanism of its neuroprotective properties. Thus, the research results available today suggest that xenon has a pronounced pleiotropic mechanism of brain protection. This is a partial blockade of NMDA receptors, and phosphorylation of the enzyme glycogen synthase-3β, and limitation of the inflammatory activation of neutrophils.Findings. Inhalation of xenon in volunteers (30% within 60 minutes) has a pronounced anti-inflammatory effect on neutrophils stimulated by lipopolysaccharides, decreasing their activation by inhibiting proinflammatory ERK 1/2 kinase and proinflammatory MAP kinase p38, as well as reducing the expression of markers of activation and degranulation CD11b and CD66b on the surface of neutrophils. Stimulation by lipopolysaccharides statistically significantly reduces spontaneous apoptosis of neutrophils, while xenon increases the ability of neutrophils to apoptosis, which is likely to contribute to the resolution of inflammation.

Infectious disease COVID-19 caused by the SARS-CoV-2 coronavirus is characterized by high contagiousness, complexity of pathogenesis and unpredictability of the clinical course. In severe cases, which are especially susceptible to men, the elderly and people with underlying medical conditions such as obesity, diabetes, hypertension, cardiovascular and chronic respiratory diseases, the infection leads to respiratory failure and death due to the development of an extensive inflammatory reaction. As a result of many studies, it has been established that one of the leading causes of the severe course and death of patients with COVID-19 is the development of coagulopathy, that is, increased thrombus formation in small vessels due to excessive activity of neutrophils, which form the so-called neutrophil extracellular traps (NETs). Although NETs play a useful role in protecting their host from pathogens, their overgrowth can trigger a cascade of adverse reactions including: the production of antibodies against the host’s DNA (autoimmunization); damage to surrounding tissue; or the occurrence of thromboembolic complications. Therefore, extracellular neutrophil traps and their markers have been identified as targets for new therapeutic strategies aimed at reducing the severity of COVID-19 disease and/or mortality. This article describes the structure of NETs, as well as analyzes the molecular mechanisms that contribute to their overgeneration. In addition, the prospects for COVID-19 therapy aimed at regulating the formation of extracellular traps by creating drugs both limiting the production of NET structures and dissolving their excess amounts in the body of patients are discussed.

Numerous studies have proven a close relationship between inflammatory diseases and the state of hypercoagulability. In fact, thromboembolic complications represent one of the main causes of disability and mortality in acute and chronic inflammatory diseases, cancer and obstetric complications. Despite this, the processes of hemostasis and immune responses have long been considered separately; currently, work is underway to identify the molecular basis for a relationship between such systems. It has been identified that various pro-inflammatory stimuli are capable of triggering a coagulation cascade, which in turn modulates inflammatory responses. Neutrophil extracellular traps (NETs) are the networks of histones of extracellular DNA generated by neutrophils in response to inflammatory stimuli. The hemostasis is activated against infection in order to minimize the spread of infection and, if possible, inactivate the infectious agent. Another molecular network is based on fibrin. Over the last 10 years, there has been accumulated a whole body of evidence that NETs and fibrin are able to form a united network within a thrombus, stabilizing each other. Similarities and molecular cross-reactions are also present in the processes of fibrinolysis and lysis of NETs. Both NETs and von Willebrand factor (vWF) are involved in thrombosis as well as inflammation. During the development of these conditions, a series of events occurs in the microvascular network, including endothelial activation, NETs formation, vWF secretion, adhesion, aggregation, and activation of blood cells. The activity of vWF multimers is regulated by the specific metalloproteinase ADAMTS-13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13). Studies have shown that interactions between NETs and vWF can lead to arterial and venous thrombosis and inflammation. In addition, the contents released from activated neutrophils or NETs result in decreased ADAMTS-13 activity, which can occur in both thrombotic microangiopathies and acute ischemic stroke. Recently, NETs have been envisioned as a cause of endothelial damage and immunothrombosis in COVID-19. In addition, vWF and ADAMTS-13 levels predict COVID-19 mortality. In this review, we summarize the biological characteristics and interactions of NETs, vWF, and ADAMTS-13, the effect of NETs on hemostasis regulation and discuss their role in thrombotic conditions, sepsis, COVID-19, and obstetric complications.

Introduction. The role of neutrophils in the initiation and progression of atherosclerosis as well as in the development of its complications has received scientific attention only in the recent years. Today, there is growing evidence to support a role of the CXCL12/CXCR4 axis in sustained inflammation during different chronic inflammatory diseases by retaining neutrophils at inflammatory sites.The aim of the study is to assess the diagnostic and prognostic significance of circulating CD62LloCXCR4 hi neutrophils in patients with carotid atherosclerosis.Materials and methods. A total of 75 patients (52% of men and 48% of women) aged 40 to 64 years were examined. None of them were diagnosed with atherosclerotic cardiovascular diseases. All the patients underwent carotid artery duplex scanning. The flow cytometry and CD16, CD11b, CD62L, CD182 (CXCR2) and CD184 (CXCR4) conjugated monoclonal antibodies were used for phenotyping and differentiation of neutrophil subpopulations.Results. Atherosclerotic plaques in carotid arteries were detected in 72% of the patients; most of the patients were diagnosed with stenosis development in more than one of the carotid arteries (CA). The elevated levels of circulating CXCR4h neutrophils were associated with the levels of total cholesterol (r = 0.377; p = 0.001), low-density lipoprotein (LDL) cholesterol (r = 0.293; p = 0.014) and triglycerides (r = 0.388; p = 0.003). The study revealed direct correlation between the circulating CXCR4 hi neutrophil count and the cumulative percentage of CA stenosis (r = 0.300; p = 0.011), including the number of stenosed CA (r = 0.291; p = 0.034). It was also found that CXCR4 hi neutrophil counts demonstrated a statistically significant increase along with the increased number of stenosed CA (p = 0.025). The ROC analysis findings show that the elevated CXCR4 hi neutrophil counts ≥260 cells/μL made it possible to diagnose stenotic lesion of 4 CAs with a sensitivity of 71.4% and specificity reaching 76.6%.Conclusion. In patients with carotid atherosclerosis, the increased count of circulating CD62LloCXCR4 hi neutrophils was associated with the increased number of stenosed CAs, while no significant changes were observed in the other examined subpopulations of neutrophil granulocytes. The increased CD62LloCXCR4 hi neutrophil count made it possible to diagnose stenotic lesion of 4 CAs with a sufficient sensitivity and specificity.