Monocyte Activation Research Articles

Rapid recruitment and IFN-I-mediated activation of monocytes dictate focal radiotherapy efficacy.

The recruitment of monocytes and their differentiation into immunosuppressive cells is associated with the low efficacy of preclinical nonconformal radiotherapy (RT) for tumors. However, nonconformal RT (non-CRT) does not mimic clinical practice, and little is known about the role of monocytes after RT modes used in patients, such as conformal RT (CRT). Here, we investigated the acute immune response induced by after CRT. Contrary to non-CRT approaches, we found that CRT induces a rapid and robust recruitment of monocytes to the tumor that minimally differentiate into tumor-associated macrophages or dendritic cells but instead up-regulate major histocompatibility complex II and costimulatory molecules. We found that these large numbers of infiltrating monocytes are responsible for activating effector polyfunctional CD8+ tumor-infiltrating lymphocytes that reduce tumor burden. Mechanistically, we show that monocyte-derived type I interferon is pivotal in promoting monocyte accumulation and immunostimulatory function in a positive feedback loop. We also demonstrate that monocyte accumulation in the tumor microenvironment is hindered when RT inadvertently affects healthy tissues, as occurs in non-CRT. Our results unravel the immunostimulatory function of monocytes during clinically relevant modes of RT and demonstrate that limiting the exposure of healthy tissues to radiation has a positive therapeutic effect on the overall antitumor immune response.

Differentiated expressed miRNAs in splenic monocyte induced by burn injury in mice.

To find potential biomarkers based on miRNA and their potential targets in splenic monocytes in burn-injured mice. Male Balb/c mice were subjected to sham or scalding injury of 15% total body surface area. Spenic CD11b+ monocytes were purified with magnetic beads. The monocytes were cultured in the presence of lipopolysaccharide. The proliferation of monocytes was detected by MTT assay, and the cytokines in the supernatant were examined by enzyme linked immunosorbent assay. The purified monocytes were also under total RNA extraction. The differential monocytic miRNAs expression between the sham and burn-injured mice was analysed by miRNA microarray. The activity of monocytes was comparable between the two groups (p > 0.05). However, monocytes from burn-injured mice secreted higher levels of tumour necrosis factor (TNF)-α and transforming growth factor-β, but lower level of monocyte chemoattratctant protein-1. A total of 54 miRNAs were differentially expressed in monocytes from burn relative to sham-injured mice (fold >3). Further quantitative reverse transcription polymerase chain reaction confirmed that the expression of miR-146a was significantly down-regulated, while miR-3091-6p was up-regulated after burn injury. Using the combination of Miranda and TargetScan softwares, we found that mir-146a may regulate 180 potential target genes including TNF receptor related factor 6 (TRAF6), interleukin-1 receptor related kinase 1 (IRAK1) and CD28. Mir-3091-6p may regulate 39 potential targets, including SOCS7 (cytokine signal transduction inhibitor 7) and ARRB2 (arrestin, β 2). The miRNAs expressed by monocytes after burn injury may be involved in the regulation of innate immune response in burn injury.

Elevated Levels of Interleukin-1β and Interleukin-10 Are Associated With Faster Lung Function Decline in People With Well-Treated Human Immunodeficiency Virus.

People with human immunodeficiency virus (PWH) have an increased risk of chronic lung diseases and chronic inflammation. We aimed to investigate if inflammatory markers and monocyte activation are associated with faster lung function decline in PWH. We included 655 PWH from the Copenhagen Comorbidity in HIV Infection (COCOMO) Study. Eligible participants were aged ≥25 years and had 2 spirometries separated by >2 years. Inflammatory markers (interleukin [IL]-1β, IL-2, IL-6, IL-10, tumor necrosis factor-α, and interferon-γ) were measured at baseline by Luminex, and soluble CD14 and soluble CD163 by enzyme-linked immunosorbent assay. Using linear mixed models, we investigated whether elevated cytokine levels were associated with faster lung function decline. The majority of PWH were males (85.2%) with undetectable viral replication (95.3%). We found a faster decline in forced expiratory volume in 1 second (FEV1) in PWH with elevated IL-1β and IL-10, with an additional decline of 10.3 mL/year (95% confidence interval [CI], 2.1-18.6; P = .014) and 10.0 mL/year (95% CI, 1.8-18.2; P = .017), respectively. We found no interaction between smoking and IL-1β or IL-10 on FEV1 decline. Elevated IL-1β and IL-10 were independently associated with faster lung function decline in PWH, suggesting that dysregulated systemic inflammation may play a role in the pathogenesis of chronic lung diseases.

Changes in <i>in vitro</i> GM-CSF-exposured monocyte subset composition and phagocytic activity in children with infectious mononucleosis

The aim of the study was to investigate the features of changes in the monocytes subset composition and phagocytic activity in children with infectious mononucleosis (IM) exposed to granulocyte-macrophage colony-stimulating factor (GM-CSF) in vitro. We examined 84 children aged 3 to 11 years with EpsteinBarr virus (EBV) infection diagnosed by clinical signs, positive EBV DNA test in blood lymphocytes and ELISA data (EBV-VCAIgM (+), EBV-EA-DIgG (+)). The control group consisted of 40 apparently healthy age-matched children. Monocytes were obtained by the standard method on adhesion to plastic from mononuclear cells isolated from heparinized venous blood by density gradient centrifugation. The isolated monocytes were divided into two samples: control (without GM-CSF) and experimental (50 ng of GM-CSF per 1 ml of cell suspension). The monocyte subset composition and phagocytic activity in both samples were measured by flow cytometry after 1-hour incubation at 37C ina CO2-incubator. It was found that in children with progressing IM, the blood monocyte subpopulation composition changes and their phagocytic activity is impaired. It was found that the subset composition and phagocytic activity of the blood monocytes changed in children during development of IM. Changes in the subset composition of monocytes in acute IM did not depend on the age group of children (36 and 711 years) and were characterized by increased number of pro-inflammatory (intermediate) monocytes and decreased level of anti-inflammatory (non-classical) monocytes. Features of altered monocyte phagocytic activity in children with IM depended on age. The phagocytic activity of all three monocyte subsets was reduced in children with IM 36 years old while children with IM 711 years old had reduced phagocytic activity only in intermediate and non-classical monocytes. The effect of GM-CSF in vitro on monocytes in patients with IM, regardless of the age of children, led to significantly increased level of anti-inflammatory monocytes while the phagocytic activity of cells changed less. An increase in the phagocytic number for classical monocytes after incubation with GM-CSF in vitro was noted in children with IM at the age of 36 years while the phagocytic index of this fraction of monocytes remained unchanged. The level of the phagocytic index increased only in classical monocytes of children with IM aged 711 years. The presented results determine the scientific and clinical value of studying the mechanisms of the effect of GM-CSF on cells of the immune system and prove that this cytokine can be used in a new immunotherapeutic strategy for the treatment of IM.

The Association of Urinary MCP-1 with Disease Activity of Lupus Nephritis

Background: The association of urinary MCP-1 (Monocyte Chemoattractant Protein-1) with disease activity in lupus nephritis (LN) has been a subject of significant interest. MCP-1 is a chemokine involved in the recruitment and activation of monocytes, which play a critical role in the pathogenesis of LN. Objective: To assess association of Urinary MCP-1 with disease activity of lupus nephritis. Method: This cross sectional study was conducted in the Department of Nephrology, Dhaka Medical College Hospital, Dhaka from January, 2017 to June, 2018. This cross sectional study was performed on 60 biopsy proven lupus nephritis patients and 30 age and sex matched apparently healthy control subjects. All the patients were recruited as per inclusion and exclusion criteria. Diagnosed SLE patients who had renal involvement and undergone renal biopsy for standard clinical indications were recruited by purposive sampling and divided into two groups of active and inactive LN as per operational definition. Results: During the study, The urinary MCP-1 was higher in active LN patients compared to the inactive group and both were higher than the level in the control. Plus, highest Sensitivity and specificity of uMCP-1 observed at cutoff value 433 pg/ml, where sensitivity was 0.967 and specificity was 0.900. Among 30 active lupus nephritis cases raised uMCP-1 was found in 29 cases and among 30 inactive lupus nephritis cases raised uMCP-1 was found in 3 cases. uMCP-1 showed very good agreement in diagnosis of lupus nephritis activity according to Kappa statistics. uMCP-1 in diagnosis of lupus nephritis activity showed accuracy, sensitivity, specificity, PPV and NPV were 0.933, 0.967, 0.900, 0.906 and 0.964 respectively. Conclusion: In conclusion, urinary MCP-1 holds great promise as a useful biomarker for the determination of lupus nephritis activity. Its association with disease activity, predictive value for treatment response and renal outcomes, and non-invasive nature make it an ...

IL-11 induces NLRP3 inflammasome activation in monocytes and inflammatory cell migration to the central nervous system

The objective of this study is to examine IL-11-induced mechanisms of inflammatory cell migration to the central nervous system (CNS). We report that IL-11 is produced at highest frequency by myeloid cells among the peripheral blood mononuclear cell (PBMC) subsets. Patients with relapsing-remitting multiple sclerosis (RRMS) have an increased frequency of IL-11+ monocytes, IL-11+ and IL-11R+ CD4+ lymphocytes, and IL-11R+ neutrophils in comparison to matched healthy controls. IL-11+ and granulocyte-macrophage colony-stimulating factor (GM-CSF)+ monocytes, CD4+ lymphocytes, and neutrophils accumulate in the cerebrospinal fluid (CSF). The effect of IL-11 in-vitro stimulation, examined using single-cell RNA sequencing, revealed the highest number of differentially expressed genes in classical monocytes, including up-regulated NFKB1, NLRP3, and IL1B. All CD4+ cell subsets had increased expression of S100A8/9 alarmin genes involved in NLRP3 inflammasome activation. In IL-11R+-sorted cells from the CSF, classical and intermediate monocytes significantly up-regulated the expression of multiple NLRP3 inflammasome-related genes, including complement, IL18, and migratory genes (VEGFA/B) in comparison to blood-derived cells. Therapeutic targeting of this pathway with αIL-11 mAb in mice with RR experimental autoimmune encephalomyelitis (EAE) decreased clinical scores, CNS inflammatory infiltrates, and demyelination. αIL-11 mAb treatment decreased the numbers of NFκBp65+, NLRP3+, and IL-1β+ monocytes in the CNS of mice with EAE. The results suggest that IL-11/IL-11R signaling in monocytes represents a therapeutic target in RRMS.

1496-P: Resatorvid, an Inhibitor of Toll-Like Receptor-4, Protects Allogenic Islets from Inflammatory and Immune Response

Purpose: Loss of islet mass by instant blood mediated inflammatory reaction (IBMIR) and immunological rejection are major hurdles currently facing the field of islet transplantation. Toll-like receptor 4 (TLR4) plays a central role in inflammation and shaping adaptive immune response. We investigated the protective effects of resatorvid (TAK-242) using in vitro models of islet transplantation. Methods: Soluble TAK-242 or TAK-242 conjugated to the surface of islets utilizing copper-free click chemistry with a cleavable chemical linker were used. Islets were mixed with allogenic blood for a period of 3 hours and plasma levels of miRNA-375 and miRNA-200c were analyzed by RT-qPCR. Allogenic peripheral blood mononuclear cells were cultured with islets. T cell and monocyte activation were monitored by flow cytometry. Cytokine and miRNA levels in culture supernatant were measured by Luminex assay and RT-qPCR respectively. Results: The presence of TAK-242 in soluble form as well as released by cleavable linker from islet surface significantly reduced the release of miRNA-375 (p<0.01) and miRNA-200c (p<0.01) indicating less islet damage. In allogenic co-culture of islets and PBMCs, TAK-242 inhibited the expression of CD69 in T cells as well as decreased (p<0.05) the expression of CD86 and CD80 in monocytes. The production of cytokines IL-1β (p<0.001), IL-6 (p=0.01), IL-8 (p<0.05), IP10 (p<0.05), MCP-1 (p<0.001) and TNFα (p=0.005) in culture supernatant was decreased in the presence of soluble TAK-242 as well as TAK-242 released from islet surfaces. Conclusions: Inhibition of TLR4 using small molecule, TAK-242, protected islets from innate damage due to IBMIR as well as adaptive responses of activation of T cells and monocytes. These results validate the future study of inhibition of TLR4 using in vivo transplant model(s) to further investigate the efficacy of TLR4 inhibition to improve islet transplant survival and function. Disclosure J.D.Mattke: None. B.Naziruddin: None. B.Kane: None.

A negative trial for vasoactive intestinal peptide in COVID-19-associated acute hypoxaemic respiratory failure

A negative trial for vasoactive intestinal peptide in COVID-19-associated acute hypoxaemic respiratory failure

Differential Activities of the Botanical Extract PBI-05204 and Oleandrin on Innate Immune Functions under Viral Challenge Versus Inflammatory Culture Conditions.

The Nerium oleander extract PBI 05204 (PBI) and its cardiac glycoside constituent oleandrin have direct anti-viral properties. Their effect on the immune system, however, is largely unknown. We used an in vitro model of human peripheral blood mononuclear cells to document effects under three different culture conditions: normal, challenged with the viral mimetic polyinosinic:polycytidylic acid Poly I:C, and inflamed by lipopolysaccharide (LPS). Cells were evaluated for immune activation marks CD69, CD25, and CD107a, and culture supernatants were tested for cytokines. Both PBI and oleandrin directly activated Natural Killer (NK) cells and monocytes and triggered increased production of cytokines. Under viral mimetic challenge, PBI and oleandrin enhanced the Poly I:C-mediated immune activation of monocytes and NK cells and enhanced production of IFN-γ. Under inflammatory conditions, many cytokines were controlled at similar levels as in cultures treated with PBI and oleandrin without inflammation. PBI triggered higher levels of some cytokines than oleandrin. Both products increased T cell cytotoxic attack on malignant target cells, strongest by PBI. The results show that PBI and oleandrin directly activate innate immune cells, enhance anti-viral immune responses through NK cell activation and IFN-γ levels, and modulate immune responses under inflamed conditions. The potential clinical impact of these activities is discussed.

Acute effects of cigarette smoke on Endothelial Nitric Oxide synthase, vascular cell adhesion molecule 1 and aortic intima media thickness

Background. Cigarette smoking could induce endothelial dysfunction and the increase of circulating markers of inflammation by activation of monocytes. This can lead to increased intima media thickness (IMT) of entire blood vessels and result in acceleration of the atherosclerosis process. However, to our knowledge, little is known about the role of cigarette smoking in this atherosclerotic inflammatory process. The aim of this study is to explore the link between cigarette smoking and its effect on endothelial nitric oxide synthase (e-NOS) and vascular cell adhesion molecule 1 (VCAM-1). Methods. An experimental study with a post-test only controlled group design was used. We used 18 Wistar rats (Rattus norvegicus) randomly subdivided into two groups: group K (-) were not exposed to tobacco smoke, whereas group K (+) were exposed to smoke equivalent of more than 40 cigarettes for 28 days daily. After 28 days, samples were analyzed for e-NOS, VCAM-1 and aortic IMT. Results. Our results indicate that tobacco smoke can enhance the expression of VCAM-1 on rat cardiac vascular endothelial cells, resulting in a decreased expression of e-NOS level and increase of aortic IMT. Linear regression model found that eNOS level negatively correlated wiith aortic IMT (r2 = 0.584, β = -0.764, p < 0.001), whereas VCAM-1 expression did not correlate with aortic IMT (r2 = 0.197, p = 0.065). Conclusion. Low e-NOS level and high VCAM-1 level observed after cigarette smoke exposure which may increase aortic IMT.

Effect of a pyrimidine derivative on the level of cytokines in conditions of streptococcal wound infection

Diseases caused by Streptococcus pyogenes are one of the significant problems of practical health care, due to the ability of this pathogen to cause morbidity in all age groups of the population with the development of different complications. The streptococcal infection is accompanied, along with infectious and inflammatory symptoms, by a pronounced immune response to Streptococcus pyogenes, which is mediated by the functional activity of monocytes and dendritic cells responsible for the production of pro- and anti-inflammatory cytokines that regulate the interaction, proliferation and functional activity of all participants in the cellular and humoral links of immunity. The purpose is to study the effect of a pyrimidine derivative on the level of pro- (IL-1β, IL-6, IL-8) and anti-inflammatory interleukins (IL-4, IL-10) in conditions of experimental streptococcal wound infection. The study of the effect of the pyrimidine derivative 3-[2-[(4,6-dimethylpyrimidin-2-yl) amino]-2-oxoethyl]quinazolin-4(3H)-one was carried out in vivo on the model streptococcal wound infection. The experiments were carried out on CBA mice 3 months old (20-22 g). Animals were divided into groups: control I–healthy animals, which were injected with water for injection; control II–infected untreated animals; experience I – animals that were treated intraperitoneally with a pyrimidine derivative at a dose of 36 mg/kg for 7 days, starting from the first day of infection; experiment II – animals that were treated intraperitoneally with the reference drug cefepime at an average therapeutic dose of 50 mg/kg in the same regimen as the test compound. The levels of pro- and anti-inflammatory interleukins in blood serum were determined by enzyme immunoassay. The study found that the pyrimidine derivative 3-[2-[(4,6-dimethylpyrimidin-2-yl)amino]-2-oxoethyl]quinazolin-4(3H)-one under conditions of streptococcal wound infection has an immunoregulatory effect, manifested in a decrease in pro-inflammatory and an increase in anti-inflammatory interleukins.

#4451 MONOCYTE MIGRATION AND ACTIVATION IN ANCA-ASSOCIATED GLOMERULONEPHRITIS

Abstract Background and Aims Glomerular injury in Anti-Neutrophil Cytoplasmic Antibodies (ANCA)-associated glomerulonephritis (AGN) is associated with macrophage infiltration. To date, their role and origin remain to be fully elucidated. Activated blood monocytes likely transmigrate and differentiate into kidney macrophages depending on the local microenvironment. Even in remission, some studies indicate sustained monocyte activation and upregulation of adhesion markers. The aim of this study was to research the intrinsic migration capacity of monocytes in AGN. Targeting monocyte migration might open new therapeutic avenues in the treatment of AGN. Method Transendothelial migration of monocytes was tested in AGN patients with active disease (n = 2), in remission (n = 8) and healthy controls (n = 6). To assess their adhesive and migratory capacity, freshly isolated CD14 positive monocytes were added to confluent TNF-α or IL1β overnight-stimulated Human Aortic Endothelial Cell (HAEC) for 30 minutes and subsequently fixed. Monocyte adhesion and transmigration was visualized by phase-contrast microscopy and quantified using ImageJ. To unravel the potential mechanisms driving these differences in monocyte migration, we performed bulk RNA-sequencing of monocytes from AGN patients with active (n = 4) and stable (n = 10) disease, and healthy controls (HC) (n = 6). Results Monocyte adhesion, but not migration, was significantly increased during active disease, independently of the stimuli used to mimic the pro-inflammatory phenotype of endothelial cells (EC) (Figure 1). During remission, decreased adhesion was found on the IL1β-stimulated ECs. While CD11b mRNA expression was upregulated, CD11a expression was downregulated in monocytes from active AGN patients compared to HC. Two genes involved in paracellular monocyte transmigration (JAML, PECAM-1) were significantly decreased. Most AGN patients were treated with corticosteroids at time of experiments. Conclusion These results suggest a remarkable increase in monocyte adhesion, but lower intrinsic migration capacity, in a 30 minute time-frame during active AGN. Our findings on migration are surprising in the light of theorized enhanced monocyte extravasation towards diseased AGN kidneys. Therefore, it could be speculated that after a longer period of time, a subset of the adherent monocytes would ultimately migrate and differentiate into kidney macrophages. In remission, long-term immunosuppressive treatment or chronic inflammation might decrease monocyte adhesion. This was the first ex-vivo study to research monocyte migration in AGN, further research is required to validate findings and to develop new therapies targeting monocyte migration.

Endothelial CCRL2 induced by disturbed flow promotes atherosclerosis via chemerin-dependent β2 integrin activation in monocytes.

Chemoattractants and their cognate receptors are essential for leucocyte recruitment during atherogenesis, and atherosclerotic plaques preferentially occur at predilection sites of the arterial wall with disturbed flow (d-flow). In profiling the endothelial expression of atypical chemoattractant receptors (ACKRs), we found that Ackr5 (CCRL2) was up-regulated in an endothelial subpopulation by atherosclerotic stimulation. We therefore investigated the role of CCRL2 and its ligand chemerin in atherosclerosis and the underlying mechanism. By analysing scRNA-seq data of the left carotid artery under d-flow and scRNA-seq datasets GSE131776 of ApoE-/- mice from the Gene Expression Omnibus database, we found that CCRL2 was up-regulated in one subpopulation of endothelial cells in response to d-flow stimulation and atherosclerosis. Using CCRL2-/-ApoE-/- mice, we showed that CCRL2 deficiency protected against plaque formation primarily in the d-flow areas of the aortic arch in ApoE-/- mice fed high-fat diet. Disturbed flow induced the expression of vascular endothelial CCRL2, recruiting chemerin, which caused leucocyte adhesion to the endothelium. Surprisingly, instead of binding to monocytic CMKLR1, chemerin was found to activate β2 integrin, enhancing ERK1/2 phosphorylation and monocyte adhesion. Moreover, chemerin was found to have protein disulfide isomerase-like enzymatic activity, which was responsible for the interaction of chemerin with β2 integrin, as identified by a Di-E-GSSG assay and a proximity ligation assay. For clinical relevance, relatively high serum levels of chemerin were found in patients with acute atherothrombotic stroke compared to healthy individuals. Our findings indicate that d-flow-induced CCRL2 promotes atherosclerotic plaque formation via a novel CCRL2-chemerin-β2 integrin axis, providing potential targets for the prevention or therapeutic intervention of atherosclerosis.

Pengaruh Pemberian Ekstrak Kulit Manggis pada Tikus Tuberkulosis yang Diberi Isoniazid terhadap Kadar Kolesterol


 
 
 
 
 
 
 
 
 
 
 
 
 Patients with tubeculosis experience a decrease in the amount of cholesterol in the body due to an inflammatory process due to increased free radical formation due to oxidative stress arising as a result of increased phagocytic activity of monocytes and macrophages. Therefore, additional treatment is needed to overcome free radicals and ROS so that inflammation in TB patients does not get worse and cholesterol levels can return to normal. Treatment of isoniazid in TB patients can increase cholesterol levels but does not occur significantly, and the results are still lower than cholesterol levels in healthy individuals. Mangosteen peel extract has a variety of ingredients and one of them is xanthones which can be anti-inflammatory and can inhibit the increase in the formation of free radicals in TB patients. The main objective of this study was to determine the effect of mangosteen peel extract on TB rats given isoniazid on cholesterol levels. This research is a laboratory experimental study with a randomized posttest only method. The research sample used by the previous researcher, Dr. dr. Indah Widyaningsih., M. Kes was the blood serum of white rats (Rattus Norvegicus) aged 2-3 months with a body weight of 200-250 grams. The analytical test used in this study is the Kruskall-Wallis test. The results of the analysis that has been done by researchers, it is known that there is no effect of Mangosteen Skin Extract on Cholesterol Levels in TB rats given Isoniazid
 
 
 
 
 
 
 
 
 
 
 
 

Comparative Analysis of Fcγ and Complement Receptors Presence on Monocytes in Pulmonary Sarcoidosis and Tuberculosis.

Sarcoidosis (SA) is a granulomatous disorder, which mostly affects the lungs. Its clinical characteristics resemble tuberculosis (TB), but its treatment is different. The etiology of SA is unknown; however, mycobacterial antigens were proposed as environmental factors in its development. Due to previously revealed immunocomplexemia with mycobacterial antigens in the blood of our SA but not TB patients, and in the search for biomarkers for differential diagnosis of the two disorders, we studied the phagocytic activity of monocytes from both patients' groups with flow cytometry. With the use of this method, we also analyzed the occurrence of receptors for IgG (FcγR) and complement components (CR) at the surface of these monocytes, responsible for phagocytosis of immunocomplexes. We revealed a higher phagocytic activity of monocytes in both disorders, but an increased frequency of monocytes with FcγRIII (CD16) and decreased with CR1 (CD35) receptor in the blood of SA vs. TB patients. With regard to our other genetic study on FcγRIII variants in SA and TB, this may account for the decreased clearance of immunocomplexes and different immune responses in the two diseases. Thus, the presented analysis not only sheds light on the pathomechanisms of SA and TB but may also support their differential diagnosis.

Phagocytic activity of peripheral blood monocytes under <i>in vivo</i> and <i>in vitro</i> hypoxia conditions in tolerant and susceptible to oxygen deficiency rats

It is known that there are individual differences in resistance to hypoxia, which can determine the predisposition to the development and severity of various diseases, including infectious, inflammatory and tumor. There are no standardized methods for assessing resistance to hypoxia in experimental animals and humans without hypoxic exposure. The search for molecular-biological markers, identifying people with different resistance to oxygen deficiency under normoxic conditions or under moderate hypoxic exposure is undoubtedly efficient. It is possible that the assessment of the basic resistance to hypoxia can help to predict the development and severity of the course of diseases, the mechanisms of which are associated with oxygen deficiency. One of the methods to assess organism resistance to hypoxia without exposure in a decompression chamber or in highland conditions can be modeling hypoxia in vitro. The aim of the study was to characterize the phagocytic activity of peripheral blood monocytes in tolerant and susceptible to hypoxia Wistar rats under normoxic conditions, as well as after hypoxic exposure in vitro and in vivo. The resistance of rats to hypoxia was determined by the gasping time at an altitude of 11.500 m in a decompression chamber. A month after determining the resistance to hypoxia, one group of rats was placed in a decompression chamber at an altitude of 5,000 m for 1 hour to simulate the hypoxic state in vivo. Blood from the tail vein of the other group of rats was placed in 1% oxygen for 1 hour to simulate the hypoxic state in vitro. The phagocytic activity of peripheral blood monocytes was assessed by flow cytometry. It was demonstrated that phagocytic activity of monocytes did not differ in tolerant and susceptible to hypoxia rats under normoxic conditions. The phagocytic activity of monocytes after in vitro and in vivo hypoxic exposure was higher in tolerant to hypoxia animals in comparison to susceptible ones. An increase in the phagocytic activity of monocytes compared to normoxia conditions was observed only in tolerant rats under in vitro conditions of hypoxic exposure. The obtained results indicate that tolerant and susceptible to hypoxia organisms differ in the phagocytic activity of monocytes under conditions of oxygen deficiency, which can determine the course of inflammatory and tumor diseases. The data obtained will be the basis for further experimental investigations organism hypoxia resistance markers.

Functional activity of the monocyte immune link in gastric adenocarcinoma

Stomach cancer is in the top ten in terms of prevalence and ranks 4th in terms of causes of death worldwide. The most common and most aggressive variant of gastric cancer is adenocarcinoma. The monocytic link of immunity provides the main line of the body's fight against malignant cells, while in patients with adenocarcinoma it is insufficiently studied. The purpose of the study was to evaluate the functional activity of monocytes in patients with gastric adenocarcinoma at different stages of the disease.Individuals (n = 164) were examined, among whom 85 were diagnosed with stage I-IV stomach adenocarcinoma. The study also involved 79 apparently healthy donors. The functional activity and oxygen-dependent phagocytosis of monocytes were assessed by the chemiluminescent method. Luminol was used as a chemiluminescence inducer. The respiratory burst was activated with opsonized zymosan.In patients with stomach adenocarcinoma, at rest (spontaneous chemiluminescence), an increase in the time the curve reached the maximum intensity of chemiluminescence (Tmax = 7957 s), the area under the chemiluminescence curve (Squr = 0.2 x 106), the activation index (1.89 c. u.) and a decrease in the maximum value of chemiluminescence intensity (Imax = 424 c. u.) relative to the control group (Tmax = 5533 s, Squr = 0.011 x 106, activation index = 0.88 c. u., Imax = 424 c. u., p < 0.05) were seen. When chemiluminescence is induced in patients with stomach adenocarcinoma, a statistically significant predominance of Squr is fixed (0.46 x 106, in the control group Squr = 0.031 x 106). Also, in the group of patients with stomach adenocarcinoma, monocytic phagocytosis was reduced by more than 2 times (29% vs 84% in the control group, p < 0.05). When analyzing the studied parameters, depending on the stage of the disease, it was found that the violation of the chemiluminescent reaction in patients with stomach adenocarcinoma is fixed already at an early stage. At the same time, in patients with stage IV stomach adenocarcinoma, the indicators of spontaneous and induced chemiluminescence are more than 2 times different from those in the control group and patients at stage I of the disease. The identified features indicate a decrease in the effectiveness of immune reactions of the monocytic link in stomach adenocarcinoma already in the early stages of the disease and can be used to detect early signs of immune disorders and optimize therapeutic approaches in this disease.

Изменения иммунологических показателей у пострадавших с политравмой при применении в комплексном лечении электромагнитных волн терагерцевого диапазона

Introduction Improving the results of treatment and rehabilitation is essential for polytrauma patients. The aim of the study was to prove the effectiveness of terahertz electromagnetic waves (TEW) in polytrauma patients early after traumatic event to improve immune parameters. Material and methods The study included 20 polytrauma patients who were divided into two groups. Patients of the treatment group (n = 10) received TEW early after traumatic event in addition to standard treatment using the damage control approach. Control patients (n = 10) received no TEW therapy in the complex of therapeutic measures. Immunological studies were performed during treatment. The effectiveness of rehabilitation was assessed at a long term using an evaluation scale. Results The mean rehabilitation score was significantly higher in the treatment group compared to the controls. The treatment group showed significantly higher production of TNFα and IL-2, spontaneous production of IFNγ by lymphocytes, phagocyte activity (NBT-test) and absorption activity of monocytes after the use of TEW in comparison with controls. An increased level of IgG and IgM was also noted in patients of the treatment group. Discussion The findings showed that the use of TEW in polytrauma patients early after traumatic event resulted in the increase in cytotoxic activity of lymphocytes and phagocytic cells. The immunostimulating effect of TEW was also achieved through boosting humoral immunity. Conclusion The TEW used early after traumatic event in polytrauma patients resulted in strengthening of the immune system which contributed to more effective rehabilitation.

Comparative analysis of the expression of the soluble IL-7 receptor in patients with arthropathy

Arthropathy is one of the most prevalent diseases, which are based on the destruction and remodeling of cartilage and bone tissue. The inflammation that precedes destruction can be caused by mechanical stress on the joints, or by autoimmune reactions. Recently, IL-7 is considered as one of the key cytokines that promote the production of matrix metalloproteinases, catabolic enzymes, T cell-mediated activation of monocytes, and maturation of osteoclasts. The soluble form of the IL-7 receptor can help prolong the lifespan of IL-7 and thereby it ensures the bioavailability of the cytokine and mediates effect of IL-7 on cells. The aim of this study was to determine the soluble form of the IL-7 receptor (sIL-7R) in the blood plasma of patients with rheumatoid arthritis (RA), osteoarthritis (OA), psoriatic arthritis (PsA) and psoriasis vulgaris (PS), as well as healthy individuals. The RA patients included in the study had moderate to high disease activity according to the DAS28 index. Patients with PsA predominantly had moderate and low disease activity (DAS28) and were characterized by mild to moderate disease severity (PASI). In accordance with the PASI index, patients with PS with mild and severe severity of the disease were included in the study. All patients with OA had a metabolic phenotype that is accompanied by an elevated body mass index.sIL-7R was determined in blood plasma by enzyme-linked immunosorbent assay. It was found that in patients with arthropathy, the level of soluble form of IL-7 was increased relative to healthy individuals, with the exception of the group of patients with PsA. Also, a high concentration of sIL-7R was observed in patients with PS. Analyzing the clinical characteristics of the patients, we found that sIL-7R levels were elevated in RA and PsA patients with high disease activity by DAS28. In addition, positive correlations were found between the concentration of sIL-7R and DAS28 in RA and PsA. In patients with PsA with moderate severity of the disease (PASI), the concentration of sIL-7R was also increased relative to donor's values. On the contrary, in patients with PS, a high level of sIL-7R was noted regardless of the severity of the disease. In patients with OA, no relationship was found between sIL-7R levels and clinical parameters.Thus, an elevated level of sIL-7R in patients with arthropathy may indicate the involvement of IL-7 and its receptor system in the pathogenesis of joint diseases. The IL-7 receptor may become a promising target both in the treatment of joint diseases and other autoimmune diseases, including psoriasis.

Synovial monocytes contribute to chronic inflammation in childhood-onset arthritis via IL-6/STAT signalling and cell-cell interactions.

Monocytes are key effector cells in inflammatory processes. We and others have previously shown that synovial monocytes in childhood-onset arthritis are activated. However, very little is known about how they contribute to disease and attain their pathological features. Therefore, we set out to investigate the functional alterations of synovial monocytes in childhood-onset arthritis, how they acquire this phenotype, and whether these mechanisms could be used to tailorize treatment. The function of synovial monocytes was analysed by assays believed to reflect key pathological events, such as T-cell activation-, efferocytosis- and cytokine production assays using flow cytometry in untreated oligoarticular juvenile idiopathic arthritis (oJIA) patients (n=33). The effect of synovial fluid on healthy monocytes was investigated through mass spectrometry and functional assays. To characterize pathways induced by synovial fluid, we utilized broad-spectrum phosphorylation assays and flow cytometry, as well as inhibitors to block specific pathways. Additional effects on monocytes were studied through co-cultures with fibroblast-like synoviocytes or migration in transwell systems. Synovial monocytes display functional alterations with inflammatory and regulatory features, e.g., increased ability to induce T-cell activation, resistance to cytokine production following activation with LPS and increased efferocytosis. In vitro, synovial fluid from patients induced the regulatory features in healthy monocytes, such as resistance to cytokine production and increased efferocytosis. IL-6/JAK/STAT signalling was identified as the main pathway induced by synovial fluid, which also was responsible for a majority of the induced features. The magnitude of synovial IL-6 driven activation in monocytes was reflected in circulating cytokine levels, reflecting two groups of low vs. high local and systemic inflammation. Remaining features, such as an increased ability to induce T-cell activation and markers of antigen presentation, could be induced by cell-cell interactions, specifically via co-culture with fibroblast-like synoviocytes. Synovial monocytes in childhood-onset arthritis are functionally affected and contribute to chronic inflammation, e.g., via promoting adaptive immune responses. These data support a role of monocytes in the pathogenesis of oJIA and highlight a group of patients more likely to benefit from targeting the IL-6/JAK/STAT axis to restore synovial homeostasis.