Functional Activity Of Macrophages Research Articles

EFFECT OF B. SUBTILIS ІМV B-7724 LECTIN ON THE ACTIVITY OF EFFECTORS OF CELLULAR ANTITUMOR IMMUNITY OF MICE WITH LEWIS LUNG CARCINOMA.

To evaluate the effect of B. subtilis IMV B-7724 lectin on the functional activity of macrophages (Mph), natural killer (NK) cells and cytotoxic lymphocytes (CTL) of mice bearing Lewis lung carcinoma (LLC). The studies were performed on C57Bl/6J mice; LLC was used as an experimental transplantable tumor. The lectin from B. subtilis IMV B-7724 was administered to LLC-bearing mice subcutaneously at a dose of 1 mg/kg of body weight for 10 days. The immunological testing was performed on days 14, 21, and 28 after tumor grafting. The cytotoxic activity of Mph, NK, and CTL was estimated in MTT-assay; the content of the stable metabolites of nitric oxide (NO) was measured by a standard Griess reaction; the arginase activity (Arg) was determined based on the measurement of urea. The administration of the B. subtilis IMV B-7724 lectin to LLC-bearing mice exerted its antitumor and antimetastatic effects partially via a significant (p < 0.05) increase of Mph and NK activities after the completion of the treatment. In the group of animals injected with lectin, the NO/Arg ratio increased significantly, indicating the prevalence of Mph with proinflammatory and antitumor properties. The cytotoxic activity of Mph exceeded the indices of untreated mice and intact control by 1.8 times and 5.3 times respectively; of NK - by 2.8 and 1.3 times respectively. The effect of treatment on the CTL activity was less pronounced. Antitumor and antimetastatic activity of the lectin from B. subtilis IMV B-7724 ensured the preservation of the cytotoxic activity of the main effectors of antitumor immunity (Mph, NK, and CTL) throughout LLC growth.

Roles of extracellular vesicles on macrophages in inflammatory bone diseases.

Inflammatory bone disease is a general term for a series of diseases caused by chronic inflammation, which leads to the destruction of bone homeostasis, that is, theosteolytic activity of osteoclastsincreases,andthe osteogenicactivity ofosteoblasts decreases, leading toosteolysis. Macrophages areinnate immunecell with plasticity,andtheirpolarization is related to inflammatory bone diseases. The dynamic balanceof macrophagesbetween theM1 phenotype and theM2 phenotypeaffectsthe occurrence and development ofdiseases. In recent years, an increasing number of studies have shown that extracellular vesicles existing in the extracellular environment can act on macrophages, affecting the progress of inflammatory diseases. This process is realized by influencing the physiological activity or functional activity of macrophages, inducing macrophages to secrete cytokines, and playing an anti-inflammatory or pro-inflammatory role. In addition, by modifying andediting extracellularvesicles, thepotentialof targeting macrophagescan beused to provide new ideasfordeveloping newdrug carriersforinflammatory bone diseases.

Influence of macrophages on the insulin-synthesizing system under normal conditions and in alloxan diabetes

Insulin-synthesizing cells (ISCs) of pancreatic gland are localized both in its islets, and in exocrine portion, as single cells or cellular agglomerates. ISCs differ in their morphological and functional characteristics, depending on characteristics of the microenvironment. Resident macrophages are also involved into formation of their microenvironment. Our purpose was to assess the effect of functional macrophages upon the insulinsynthesizing system (pancreatic islets, cell agglomerates, and separately lying insulin-synthesizing cells) under normal conditions and in alloxan diabetes.Alloxan diabetes was induced in mature male Wistar rats by intraperitoneal injection of alloxan (30 mg/100 g). Functional activity of macrophages was modeled with anti-inflammatory drug aminophthalhydrazide (AMP). Contents of insulin, glucose, and glycosylated hemoglobin were measured in blood of experimental animals. The levels of IL-1α, TNFα and IFNγ were determined in pancreatic homogenate. The number of macrophages was counted in histological preparations from the insular and exocrine parts of the organ, as well as the number of pancreatic islets, agglomerates, and single ISCs. The amounts of proliferating cells (insulin+Ki-67+), apoptotic forms (TUNEL+insulin+), and insulin content of ISCs at different sites (according to their fluorescence intensity) were determined. All pancreatic islets were divided into 3 types, according to intensity of insulin fluorescence, i.e., islets with high, median and low levels of fluorescence.In healthy rats, immunomodulation reduced total level of IL-1α in pancreatic parenchyma, without changing the overall parameters of carbohydrate metabolism. In the exocrine part of pancreas, the content of single ISCs in ductal epithelium was increased. Likewise, proliferation of the ISC agglomerates became higher. The intensity of β-cell apoptosis increased in pancreatic islets. The proportion of islets with high-level insulin fluorescence was decreased, along with lower density of macrophages and proliferation rates of β-cells, and higher apoptosis rates, than in intact animals. We have also revealed there an increased ratio of cells with average insulin levels. In the islets with low insulin content, immunomodulation did not cause morphological changes. Administration of AMP in alloxan diabetes contributes to a significantly decreased concentration of IFNγ in pancreatic tissues, stabilizes IL-1α content, along with reduced apoptosis of ISCs and macrophage infiltration in all parts of the gland. In the ductal epithelium, a large number of single ISCs with high synthetic activity was observed, with retained number of agglomerates and their increased cellularity. The number of dividing β-cells is increased in pancreatic islets.Modulation of the functional activity of pancreatic macrophages under physiological conditions provides a multidirectional effect on the insulin-synthesizing cells, depending on their localization. In exocrine part of the organ, where M2 macrophages are located, we have observed activated differentiation and proliferation of ISC precursors. Meanwhile, in the islets where M1 macrophages are present, apoptosis of β-cells was enhanced. In alloxan diabetes, immunomodulation was associated with reduced destruction of insulinocytes, along with high intensity of their proliferation. Heterogenous response of ISCs to the changes in the microenvironment depends on their synthetic activity. In healthy rats, the islets with high level of insulin fluorescence, the level of apoptosis is increased, and β-cell proliferation is reduced, while the morphological and functional characteristics of islets with low-level insulin fluorescence did not change. In alloxan diabetes, apoptosis prevailed in islets with high fluorescence values, whereas β-cell proliferation predominated in the islets with low insulin contents.

Abstract 1362: Impact of chemotherapy on functional and biochemical characteristics of cancer-associated fibroblasts​

Abstract Cancer-associated fibroblasts (CAFs), as the major components of the tumor microenvironment (TME), have been extensively explored and are known to be involved in tumor biology, including growth, energy metabolism, immune regulation, invasion and metastasis. A large number of preclinical studies describe the functional and biochemical impact of commonly used chemotherapeutics, such as paclitaxel and oxaliplatinum, on cancer cells, but their effect on the phenotypical and biochemical changes of CAFs and their reciprocal impact on tumor progression are lacking. In our study, we used patient-derived breast and colorectal CAF to evaluate dose-response effects of paclitaxel and oxaliplatinum, respectively first-line chemotherapeutics in breast and colorectal cancer. We evaluated the damage responses of CAF and investigated changes in morphology, cell death, woundhealing migration, type I collagen contraction and invasion, cytokine and chemokine release, differential protein expression, and metabolic activity. In response to paracrine signals emerging from chemotherapy-treated CAF we investigated the impact on functional activity of macrophages, neutrophils and cancer cells. Our project could help us better understand and modulate the activity of CAFs, which might provide novel targets for the clinical treatment of cancer. Citation Format: Zihan Xia, Olivier De Wever. Impact of chemotherapy on functional and biochemical characteristics of cancer-associated fibroblasts​ [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1362.

Protective effect of soloxolone derivatives in carrageenan- and LPS-driven acute inflammation: Pharmacological profiling and their effects on key inflammation-related processes

The anti-inflammatory potential of three cyanoenone-containing triterpenoids, including soloxolone methyl (SM), soloxolone (S) and its novel derivative bearing at the C-30 amidoxime moiety (SAO), was studied in murine models of acute inflammation. It was found that the compounds effectively suppressed the development of carrageenan-induced paw edema and peritonitis as well as lipopolysaccharide (LPS)-driven acute lung injury (ALI) with therapeutic outcomes comparable with that of the reference drugs indomethacin and dexamethasone. Non-immunogenic carrageenan-stimulated inflammation was more sensitive to the transformation of C-30 of SM compared with immunogenic LPS-induced inflammation: the anti-inflammatory properties of the studied compounds against carrageenan-induced paw edema and peritonitis decreased in the order of SAO > S > > SM, whereas the efficiency of these triterpenoids against LPS-driven ALI was similar (SAO ≈ S ≈ SM). Further studies demonstrated that soloxolone derivatives significantly inhibited a range of immune-related processes, including granulocyte influx and the expression of key pro-inflammatory cytokines and chemokines in the inflamed sites as well as the functional activity of macrophages. Moreover, SM was found to prevent inflammation-associated apoptosis of A549 pneumocytes and effectively inhibited the protease activity of thrombin (IC50 = 10.3 µM) tightly associated with rodent inflammatome. Taken together, our findings demonstrate that soloxolone derivatives can be considered as novel promising anti-inflammatory drug candidates with multi-targeted mechanism of action.

The effect of enzymatic hydrolysate of lymphoid tissue of broiler chickens on the morphofunctional state of macrophages

The goal of studying the mechanisms of action of cells that structure the immune system is to obtain additional data about the influence of immunomodulating peptides of natural origin on the possibility of regulating immune processes. The effect of enzymatic hydrolysate of lymphoid tissue of broiler chickens (EHLTBC) on the viability of culture of peritoneal and alveolar macrophages, on the morphological characteristics and functional activity of macrophages has been studied. The effect of EHLTBC on the viability of culture of peritoneal and alveolar macrophages has been investigated. Morphometric characteristics of macrophages under the influence of EHLTBC have been given. The functional activity of peritoneal and alveolar macrophages has been assessed against the background of the introduction of EHLTBC. Enzymatic hydrolysate of lymphoid tissue of broiler chickens at concentrations of 1, 5 and 10 % does not have a toxic effect on the culture of alveolar and peritoneal macrophages. EHLTBC enhances the phagocytic activity of peritoneal macrophages, which enhances their role in the chemotaxis and immune response. The stimulating effect of EHLTBC of poultry on the ability of macrophages to adhere has been revealed.

THE EFFECT OF B. subtillis ІМV B-7724 LECTIN ON FUNCTIONAL ACTIVITY OF THE MAIN EFFECTORS OF ANTITUMOR IMMUNITY OF INTACT MICE

Aim. The purpose of the study was to evaluate the effect of B. subtilis IMV B-7724 lectin on the functional activity of macrophages (Mph) and natural killer cells (NK) of intact Balb/c mice. Materials and Methods. Balb/c mice were subjected to 10 consecutive administrations of the lectin in a dose of 1 mg/kg of body weight. The functional activity of peritoneal Mph and NK were studied. Statistical analysis of the results was performed according to the widely accepted methods of variational statistics. Results. Administration of bacterial lectin increased Mph and NK cytotoxic activity; maximal increase was registered after the complete course of injections. A significant increase in the NO production indicates the polarization of peritoneal Mph into pro-inflammatory type M1. The transcription factors of IRF (at the early stage) and STAT (at the latter stages) signalling pathways were involved in the process of Mph polarization. Conclusion. The ability of B. subtillis IMV B-7724 lectin to increase in vivo cytotoxic activity of innate immunity effectors and to maintain the long-term Mph M1 polarization urges further studies on the lectin effectiveness.

TRPA1-mediated effects on the functional activity of macrophages under the exposure with cigarette smoke and cinnamaldehyde

Introduction. Being the leading cause of COPD, smoking represents a major health problem. Upon entering the respiratory tract, cigarette smoke comes into contact with various cells, including macrophages expressing on their surface TRPA1 receptors, which are sensitive to the main pathogenic compounds formed during tobacco combustion.Aim. To study the functional activity of TRPA1 channels on macrophages in terms of cell responses to cigarette smoke and the TRPA1 agonist cinnamaldehyde (CA). Materials and methods. The experimental conditions included exposure of monocyte-derived macrophages to CA (100 μM), 4% cigarette smoke extract (CSE) and 4% CSE after pretreatment with TRPA1 selective antagonist (HC-030031 100 μM). The concentration of cytokines in the culture medium, the expression of TRPA1 on the cell surface, as well as the phagocytic activity of macrophages were analyzed by flow cytometry.Results. We found that 60.2 (49.6; 71.8)% of cells expressed TRPA1 and their number increased after exposure with CA. CSE significantly inhibited CXCL10 production from 1121.3 (295.7; 3154.6) pg/ml to 187.9 (113.8; 398.3) pg/ml (p=0.04), which was partially prevented by blocking TRPA1 (692.4 [428.6; 2916.6] pg/ml, p=0.04). Similar to CSE, CA also caused a decrease in CXCL10 concentration (189.2 [111.7; 311.3] pg/ml, p=0.03). Among other observations, there was an increase in the concentration of IL-1β after the exposition with HC-030031, as well as a decrease in TNF-α, IFN-γ and IL-12p70 after the treatment with CA. CSE caused a minor inhibition in phagocytic cells number, which was not prevented by TRPA1 blocking. CA, on the contrary, increased the phagocytic activity of macrophages. The initial expression of TRPA1 had a negative correlation with the dynamics of CXCL10 in response to CSE and CA but a positive correlation with the number of phagocytic cells after exposition with CA (ρ=0.81, p=0.005). Conclusions. TRPA1 expressed on macrophages apparently mediate an anti-inflammatory effect in terms of produced cytokines but increase phagocytic activity of the cells. TRPA1 are also major receptors involved in the diminished CXCL10 production by macrophage under exposition with cigarette smoke

Experimental study on disorders in the activity of macrophages in the acute period of different severities of cerebral stroke

BACKGROUND: The development of immunosuppression in the acute phase of cerebral stroke may lead to infectious and inflammatory complications. However, data on the functional activity of macrophages are insufficient to understand the pathogenesis of stroke.&#x0D; AIM: This study aimed to investigate the functional activity of macrophages in the acute period of experimental cerebral stroke (ECS) of different severities.&#x0D; MATERIALS AND METHODS: In an experimental study on 45 Wistar rats, the functional and spontaneous metabolic activity of macrophages in the spleen and abdominal cavity and the induced metabolic activity of peritoneal macrophages were evaluated through the reduction of nitro blue tetrazolium via spectrophotometry. The spontaneous and induced adhesive activity and the inhibition of the modified adhesive activity of peritoneal macrophages were examined via crystal violet staining by using a colorimetric method. ECS was modeled by the rotational movements of mandrel-knife inserted into the region of capsula interna (c.i.) on the left by guiding a needlecannula with the subsequent damage of vessels and the formation of mild (local incision of vessels in the region of c.i.), moderate (additional introduction of autologous blood in the area of c.i.), and severe (cutting of vessels from c.i. to pia mater with the subsequent introduction of autologous blood into the damaged area) hemorrhage. Data were evaluated in 72 h after different severities of ECS were modeled.&#x0D; RESULTS: In 72 h after ECS with different severities was modeled, the functional and spontaneous metabolic activity of splenic macrophages significantly decreased compared with those in the control group. The functional, spontaneous, and induced metabolic activity of peritoneal macrophages significantly reduced compared with those in the control group. The adhesive activity of peritoneal macrophages significantly increased compared with that of the control group. These disorders exacerbated in animal models with a more severe degree of stroke.&#x0D; CONCLUSION: In the acute period of ECS, the functional activity of macrophages decreased, whereas their adhesive activity increased. These phenomena enhanced in more severe models of stroke.

Role of MP-4 myelopeptide in regulation of antibody production and functional activity of macrophages under stress conditions

Myelopeptides are endogenous low-molecular weight peptides with immunoregulatory activity. Each of these myelopeptides has an individual sequence and plays a role in correction of immune system disorders. In our earlier studies, we have shown immunomodulatory effect of MP-3, MP-5, MP-6 myelopeptides upon functioning of peritoneal macrophages under the conditions of immobilization and cold stresses. The aim of this work was to investigate the effect of MP-4 myelopeptide upon antibody formation, production of reactive oxygen species, and absorptive activity of peritoneal macrophages in mice under the immobilization stress. Materials and methods. The in vivo experiments were carried out in 24 male Swiss mice weighing 17-22 g. Two-or six-hour immobilization stress was produced in the animals. MP-4 was injected intraperitoneally 30 min before the onset of stress, at a dose of 40 μg/kg. The animals were divided into 4 groups: 1, control; 2, stress; 3, stress + MP-4; 4, MP-4 injection. On day +5, cellularity and the number of antibody-forming cells (AOC) in spleen were assessed by the method of local hemolysis in an agarose gel according to Jerne. Production of reactive oxygen species by peritoneal cells was assessed using the luminol-dependent chemiluminescence (LZHL) reaction. The absorption capacity of peritoneal macrophages was assessed using a standard method using a BDFACSCalibur laser flow cytometer. Labeled St. aureus at a final concentration of 108 cells/ml were used as phagocytosis targets. Statistical analysis was performed using the unpaired Student's t-test. Results. It was found that two-hour immobilization stress did not affect the absorption activity of peritoneal leukocytes, whereas six-hour stress suppressed phagocytosis. Injection of MP-4 in stress-exposed animals did not lead to changes in phagocytic activity of peritoneal macrophages. MP-4 did not correct the stress-induced decrease in the number of antibody-forming cells from the spleen during six hours of immobilization, nor stress-induced increase in absolute amounts of AOC under the two-hour immobilization stress. In the group of animals receiving MP-4 and immobilized for two-hours, an increase in the number of nucleated cells was increased, as compared with the control group. Thus, in contrast to the previously studied myelopeptides, MP-4 did not show pronounced immunomodulatory effects upon these parameters.

Immunophenotypic Characteristics of Macrophages in Liver and Peritoneal Area of Animals with Model of Diabetes Mellitus Type 1 and Their Corrections with Sodium Aminodihydrophthalazindione In Vitro

The goal of this study was to investigate the morphological and functional characteristics of macrophages of various localizations isolated from intact animals and animals with the model of diabetes mellitus type 1 (DM1) stimulated for 24 and 72 h in culture with sodium aminodihydrophthalazinedione (ADPN), a macrophage modulator drug. The experiments have been performed on cultures of rat macrophages obtained from the liver and peritoneal cavity. The following morphometric parameters were determined: the area of the cell, cytoplasm and nucleus, as well as the nuclear–cytoplasmic ratio (NCR). The phenotype of macrophages was determined by the expression of CD163 (macrophages M2) and CD80 (macrophages M1) markers. The functional activity of macrophages was assessed by the level of cytokines IL-1α, IL-10, and TNFα. It was found that ADPN altered the morphometric parameters (the size of cells and nuclei decreased, while NCR increased) and synthetic activity of cells (increased IL-10 in macrophages of the peritoneal cavity and IL-1α and TNFα in liver macrophages) during the first 24 h of cultivation. After 72 h, ADPN caused a decrease in the levels of IL-10 and TNFα and an increase in the level of IL-1α in all studied cell populations. ADPN did not affect the expression level of macrophage markers M1 and M2.

Natural Phenolic Acid, Product of the Honey Bee, for the Control of Oxidative Stress, Peritoneal Angiogenesis, and Tumor Growth in Mice.

Tumor-associated macrophages (TAM) are key regulators of the link between inflammation and cancer, and the interplay between TAM and tumor cells represents a promising target of future therapeutic approaches. We investigated the effect of gallic acid (GA) and caffeic acid (CA) as strong antioxidant and anti-inflammatory agents on tumor growth, angiogenesis, macrophage polarization, and oxidative stress on the angiogenic model caused by the intraperitoneal (ip) inoculation of Ehrlich ascites tumor (EAT) cells (2.5 × 106) in Swiss albino mouse. Treatment with GA or CA at a dose of 40 mg/kg and 80 mg/kg ip was started in exponential tumor growth phase on days 5, 7, 9, and 11. On day 13, the ascites volume and the total number and differential count of the cells present in the peritoneal cavity, the functional activity of macrophages, and the antioxidant and anti-angiogenic parameters were determined. The results show that phenolic acids inhibit the processes of angiogenesis and tumor growth, leading to the increased survival of EAT-bearing mice, through the protection of the tumoricidal efficacy of M1 macrophages and inhibition of proangiogenic factors, particularly VEGF, metalloproteinases -2 and -9, and cyclooxygenase-2 activity.

Effect of temperature on functional activity of macrophages in three different species

Abstract Temperature affects body physiological functions, including immunity in ways that influence survival. This study investigated the effect of temperature on macrophage activation and metabolism across species. Macrophages were isolated from mice, chicken or rainbow trout primary tissue or from macrophage cell lines and were activated with lipopolysaccharide (LPS) or IFN-γ. For mice, both Raw264.7 and primary cells, nitric oxide (NO) production was similar at 35°C and 37°C but dropped dramatically at temperatures below 35°C. At fever temperature (39°C), NO release increased in response to LPS. Young bone marrow derived macrophage (BMDM) and peritoneal resident macrophage (PRM) showed increased protein synthesis at 39°C compared to 37°C. Chicken splenic macrophages (CSM) showed NO responses that were similar at 37, 39, and at 41°C (normal for avian). A fever of 42°C had a large stimulatory effect on NO production compared to 41°C. A chicken liver derived macrophage cell line, HTC, showed the same pattern. They also showed higher protein synthesis at 42°C compared with (41°C) after LPS stimulation. Trout head-kidney macrophages (THM) showed the highest NO responses at 19°C when compared to more typical stream temperatures of 13, 15, and 16°C. They still showed some response at 28 and 37°C. Thus, their macrophages respond at higher temperatures than the fish can tolerate. Stimulated THM cells with LPS at 19°C showed increased protein synthesis compared to 13 and 16°C. We can conclude from our experiments that fish macrophages had a much broader range of temperatures at which they could respond by NO generation and protein synthesis compared to mice and chickens and all had increased responses at fever temperatures.

Антигистаминные препараты оказывают влияние на функциональную активность макрофагов

Антигистаминные препараты оказывают влияние на функциональную активность макрофагов

Macrophages and Their Role in Destabilization of an Atherosclerotic Plaque

The modern data on structure and the functional activity of macrophages are presented in the review. It is shown that they arethe nonhomogeneous cell population. Two of their main subpopulations are presented as M1 and M2 phenotypes which performopposite functions at inflammation development. The main attention in the review is paid to a role of macrophages in pathogenesisof atherosclerosis and, first, in formation of unstable atherosclerotic plaques which are the cause of the most severe complicationsof the disease. It is shown that main subpopulations of macrophages play different roles in formation of unstable and stable atheroscleroticplaques. Macrophages of M1 phenotype in the vascular wall carry out pro-atherogenic role and influence destabilizationof an atherosclerotic plaque, while M2 macrophages perform atheroprotective function.

Plasticity of Human THP-1 Cell Phagocytic Activity during Macrophagic Differentiation.

Studies of the role of macrophages in phagocytosis are of great theoretical and practical importance for understanding how these cells are involved in the organism's defense response and in the development of various pathologies. Here we investigated phagocytic plasticity of THP-1 (acute monocytic human leukemia) cells at different stages (days 1, 3, and 7) of phorbol ester (PMA)-induced macrophage differentiation. Analysis of cytokine profiles showed that PMA at a concentration of 100nM induced development of the proinflammatory macrophage population. The functional activity of macrophages was assessed on days 3 and 7 of differentiation using unlabeled latex beads and latex beads conjugated with ligands (gelatin, mannan, and IgG Fc fragment) that bind to the corresponding specific receptors. The general phagocytic activity increased significantly (1.5-2.0-fold) in the course of differentiation; phagocytosis occurred mostly through the Fc receptors, as shown previously for M1 macrophages. On day 7, the levels of phagocytosis of gelatin- and Fc-covered beads were high; however, the intensity of ingestion of mannan-conjugated beads via mannose receptors increased 2.5-3.0-fold as well, which indicated formation of cells with an alternative phenotype similar to that of M2 macrophages. Thus, the type and the plasticity of phagocytic activity at certain stages of macrophage differentiation can be associated with the formation of functionally mature morphological phenotype. This allows macrophages to exhibit their phagocytic potential in response to specific ligands. These data are of fundamental importance and can be used to develop therapeutic methods for correcting the M1/M2 macrophage ratio in an organism.

Features of the functional activity of macrophage link of immunity with gastroesophageal reflux disease depending on the type of reluctate: in vitro model.

A generalized analysis of changes in functional activity of macrophages on the basis of phagocytic activity, cytokine profile, changes in the level of expression of surface markers characteristic of pro- or anti-inflammatory phenotype of the cells when exposed to reluctate. Developed in vitro model of co-peritoneal macrophages of mice With57/BL6 (n=65) and reluctate patients with gastroesophageal reflux disease (GERD; n=65) having different pH values (three group comparison). Took into account the standard criteria phagocytic ability (absorption Staphylococcus aureus 9198, light microscopy), secretory activity (cytokine profile Th1/Th2, flow cytometry) and receptor characterization of macrophages (expression of CD25/80/163/206, flow cytometry). The phagocytic activity of macrophages, calculated on the basis of the average number of bacteria ingested by one phagocyte, is not associated with the pH value of the added reluctate. It is established that the alkalinisation of reluctate leads to significant alteration in the expression of CD receptors - decrease M1 and increase M2. The index of total production of Th1/Тһ2 in groups progressively decreased with increasing pH of reluctate and amounted to 3.6 units in the group pH from 4.6 to 6.6; 2.8 units group a pH of 6.7-7.2 and 1.6 units in the group pH of 7.3 to 8.1, due to increased production of Th2 cytokines at offset reluctate pH to slightly alkaline side. The data obtained indicate the increase of expression and secretion of anti-inflammatory markers at an alkaline pH shift of reluctate. Analysis of the studied characteristics of the activity profile of macrophages in the proposed in vitro model justifies the need for considering the peculiarities of the functional activity of macrophages under the influence of reluctate different nature. The special importance of studying the cytokine profile and characteristics of the functional activity of macrophages in patients with GERD, given the nature of reluctate.

Oxidative stress and inflammation caused by n-Hexyl salicylate in mouse skin: the effectiveness of flavonoids

&lt;p&gt;Reactive oxygen species (ROS) play a role in a numbered of degenerative conditions including psoriasis. Psoriasis is a chronic inflammatory disease who’s the etiopathogenesis is not yet completely understood, and therefore there is no standardized therapeutical approach. Flavonoids, recognized as potent antioxidants, are multifunctional molecules that can act as anti-inflammatory and antiproliferative agents through the modulation of multiple signaling pathways. The present study was designed to investigate the protective role of flavonoids [quercetin, chrysin, curcumin or Epigallocatechin 3-gallate (EGCG)] against n-Hexyl salicylate (HXS)-induced oxidative stress and inflammation in skin. Anti-oxidative and anti-inflammatory effect of flavonoids is quantified by histopathological assessment of skin, measuring the levels of lipid peroxidation and glutathione (GSH) in the skin, total number of inflammatory cells in peritoneal cavity, macrophage spreading index, and hematological and biochemical parameters.&lt;/p&gt;&lt;p&gt;Topically applied of n-Hexyl salicylate caused significant increase in lipid peroxidation and decrease in GSH, which is accompanied by an increase&lt;strong&gt; &lt;/strong&gt;&lt;em&gt;total number of inflammatory cells in skin and peritoneal cavity, functional activity of macrophages, and enzymatic activity of ALP and AST.&lt;/em&gt; In contrast, topically applied 5 % preparation of flavonoids (quercetin, chrysin, curcumin or EGCG) with HXS effectively prevented these alterations and maintained the antioxidant status.&lt;/p&gt;&lt;p&gt;The results suggest that flavonoid preparations can serve as a potent antioxidant and anti-inflammatory agents in psoriatic-like skin lesions, without toxic effects.&lt;/p&gt;

Influence of Humic Acids Extracted from Peat by Different Methods on Functional Activity of Macrophages in Vitro.

We studied activation of macrophages with humic acids extracted from peat of large deposits in the Tomsk region by two extraction methods: by hydroxide or sodium pyrophosphate. Humic acid of lowland peat types containing large amounts of aromatic carbon, phenolic and alcohol groups, carbohydrate residues and ethers, irrespectively of the extraction methods contained LPS admixture that probably determines their activating properties. Humic acid of upland peat types characterized by high content of carbonyl, carboxyl, and ester groups enhance NO production and reduce arginase expression, but these effects were minimized when sodium hydroxide was used as an extraction solvent. Pyrophosphate samples of the upland peat types were characterized by aromaticity and diversity of functional groups and have a significant advantage because of they induce specific endotoxin-independent stimulating action on antigen presenting cells.

High-density lipoprotein immunomodulates the functional activities of macrophage and cytokines produced during ex vivo macrophage-CD4+ T cell crosstalk at the recent-onset human type 1 diabetes

BackgroundBoth CD4+ T cells and macrophages are mainly involved in the autoimmune-mediated β-cells destruction in type 1 diabetes (T1D). The aim of this study was to examine the effect of HDL on functional activities of macrophage and its ability to regulate the production of cytokines in autologous mixed macrophage/CD4+ T cells at the recent-onset human type 1 diabetes. MethodsCell samples were isolated from volunteers with recent-onset T1D or healthy controls. ResultsThe levels of the production of IL-1β, IL-2, IFN-γ, nitric oxide (NO), and hydrogen peroxide (H2O2) were significantly increased in the co-culture of T1D cells when compared to that of cells from healthy controls. Similarly, those of intracellular free calcium ions (ifCa2+) were slightly, but not significantly increased (p> 0.05). Conversely, macrophage exhibited significantly decreased levels of the relative tyrosine phosphorylation of STAT6 (p-STAT6, Tyr641) in culture of T1D cells than in that of cells from healthy controls; while those of p-STAT4 (Tyr693) were significantly increased. Likewise, the levels of IL-4 and IL-10 were significantly decreased in the co-culture of T1D cells compared to co-culture of cells from healthy controls. Additionally, HDL treatment significantly down-regulated the production of IL-1β, IL-2, IFN-γ, NO, H2O2, phagocytosis, bacterial killing, the relative tyrosine phosphorylation of macrophage-expressed STAT4 (p-STAT4, Tyr693), as well as the ratio of IL-1β/IL-10, NO production/arginase activity, p-STAT4/p-STAT6, IFN-γ/IL-4, IFN-γ/IL-10, and the combined proinflammatory (PICs)/anti-inflammatory (AICs) cytokines. Moreover, HDL treatment significantly up-regulated the production of IL-4, IL-10, arginase activity, and p-STAT6 (Tyr641) (for all comparisons, p< 0.001). ConclusionsWe show for the first time that HDL may reverse both the functional activities of macrophages and immunoinflammatory response during reciprocal macrophage-CD4+ T cell crosstalk at the beginning of T1D. These findings should open the way for therapeutic trials in the short- and medium-term.