Combined Effect of TLR2 Ligands on ROS Production by Mouse Peritoneal Macrophages.

TLR2-mediated ROS production by mouse peritoneal macrophages was studied by luminoldependent chemiluminescence under conditions of cell stimulation with zymosan (TLR2/6 ligand) and peptidoglycan (TLR2/1 ligand). ROS production by macrophages stimulated with zymosan and peptidoglycan simultaneously depended on the ratio of ligand concentrations. Three effects were revealed: additivity of the stimulating effects of the ligands used, competitive ligand binding, and effect of macrophage priming with peptidoglycan during cell stimulation with zymosan. The mechanisms of these effects are discussed.

[Study of cytokine profile in men with hypertension.]

Gender differences in the risk factors' influence on the development of cardiovascular diseases determine the study of the pathogenetic causes of these features. Inflammatory is one of the mechanisms for the participation of risk factors in the development of hypertension. The aim of the study was to investigate the relationship between inflammatory biomarkers with hypertension and cardiovascular risk factors in men. The study included 60 men with hypertension who were divided into groups depending on the risk of cardiovascular events: 30 men with low risk and 30 men with high risk. 86 hypertensive patients with risk factors such as obesity, impaired glucose metabolism, impaired lipid metabolism also were examined. Biochemical parameters and a wide range of cytokines: IL-4, IL - 8, IL-10, IL-12, IL-17, IL-18, TGF-ß, TNF-α, IFN-γ were estimated. The obtained results were compared with echocardiographic (EchoCG), ambulatory blood pressure monitoring (ABPM), duplex scanning of the carotid arteries. A direct correlation between the risk factors and target organs' damage - left ventricular hypertrophy, increased intima-media complex thickness, was revealed. The dependence of cytokine profile from risk of cardiovascular events in men was also revealed. We studied the contribution of individual risk factors in men in the development of hypertension and the severity of inflammation. More significant changes in the cytokine profile observed in patients with concomitant hyperglycemia, obesity and dyslipidemia. A correlation between cytokine profile and duration of hypertension also revealed. Correlation analysis revealed the relationship between ABPM parameters and levels of interleukins, HS-CRP, and fibrinogen. Biomarkers of inflammation associated with the severity of visceral obesity, with a BMI. Analysis of results also showed the relationship between hyperinsulinemia or insulin resistance and inflammatory biomarkers. The relationship of inflammation with lipid metabolism in hypertensive men was more significant than with the other studied risk factors.

Effect of lornoxicam therapy on expression of TLR2 and TLR4 mRNA during systemic complications of acute pancreatitis.

Primary pancreatic injury that occurs in acute pancreatitis leads to necrosis of pancreatic cells and is accompanied by the development systemic inflammatory response of varying severity. Systemic inflammatory response, in turn, can lead to the development of multiple organ dysfunction syndrome and death of patients. The release of damage-associated molecular patterns into the extracellular space is the trigger pathological mechanism underlying these processes. The released patterns exert their effects via Toll-like receptors (TLR). These findings suggest that TLR can be considered a new target for therapeutic intervention in acute pancreatitis. We studied mRNA expression of TLR2 and TLR4 in the peripheral blood mononuclear cells from the patients with acute pancreatitis and showed a decrease in the examined parameters associated with lornoxicam treatment. Anti-mediator therapy decreased mortality in these patients.

[Recognition receptors of innate immunity (NLR, RLr, and CLR)].

Current concepts of recognition receptors of innate immunity (pattern-recognition receptors, PRR) are discussed in the review. Structural and functional features of receptors from the families NOD-like receptors (NLRs), RIG-like receptors (RLRs), and C-type lectin-like receptors (CLRs) are described. These receptors are found on cell surface or in cytoplasm, and also could be presented in organism in secretory form. Data on exogenous and endogenous ligands, signal transduction mechanisms that induce production of proinflammatory cytokines, chemokines, and antimicrobial petides are summarized in the review. Special attention is paid to family of NLR receptors, which are involved in generation and activation of multimeric protein complex called an inflammasome. Activation of inflammasome leads to generation of active forms of proinflammatory cytokines belonging to IL-1 family (IL-1beta, IL-18, and IL-33) from their precursor peptides due to effect of caspase-1. Data regarding involvement of innate immunity receptors in development and pathogenesis of various diseases are presented.

[The systemic inflammatory response syndrome correction in acute destructive pancreatitis].

Acute pancreatitis is a disease of variable severity. In which some patients experience mild, self-limited attacks while others manifest a severe, highly morbid, and frequently lethal attack. The exact mechanisms by which diverse etiological factors induce an attack are still unclear. Recent studies have established the role played by inflammatory mediators in the pathogenesis of acute pancreatitis. In our research we have estimated influence of not steroid anti-inflammatory preparation on synthesis pro-and anti-inflammatory Cytokines at healthy donors and at patients with Acute pancreatitis.

[Toll-like receptor-mediated functional activity of mononuclear cells in children with neutropenia].

AIM: To study the Toll-like receptor (TLR)-mediated functional activity of peripheral blood mononuclear cells (PBMC) from children with different forms of neutropenia. MATERIALS AND METHODS: TLR-mediated functional activity of PBMC was evaluated by production of proinflammatory cytokines--TNF alpha and IFN alpha. Ligands for TLR1/2, TLR 2/6, TLR4, TLR5, and TLR9 were used to stimulate TNF alpha production by PBMC from healthy children and children with neutropenia. Ligands for TLR3, TLR4, TLR7, TLR7/8, TLR8, and TLR9 were used to induce production of IFN alpha. Levels of TNF alpha and IFN alpha were measured in PBMC supernatants by ELISA. The group of patients with neutropenia included 9 children with immune neutropenia and 3 children with congenital neutropenia. Control group consisted of 12 healthy children of the same age range. RESULTS: It was revealed that TLR2, TLR4, and TLR5 ligands have enhanced stimulating effect on TNF alpha production by PBMC of children with congenital neutropenia and had no effect on PBMC of children with immune neutropenia. Children with immune neutropenia are characterized by significantly increased IFN alpha production induced by ligands of TLR3, TLR8, and TLR9. CONCLUSION: Revealed changes of TLR-mediated functional activity of PBMC from children with various forms of neutropenia may play significant role in development and course of infections in these patients.

[Corrective effect of cyclooxygenase inhibitor on functional status of mononuclear cells expressing Toll-like receptors].

AIM: To study the influence of the COX inhibitor--lornoxicam (LX)--on Toll-like receptor (TLR)-mediated production of proinflammatory and anti-inflammatory cytokines by peripheral blood mononuclear cells (PBMC) from healthy subjects and patients with acute pancreatitis (AP) in vitro. MATERIALS AND METHODS: Cytokine production by PBMC of healthy donors was stimulated by TLR1/2 ligand peptidoglycan (PG) and TLR4 ligand lypopolysaccharide (LPS) in presence of LX. Levels of cyotokines (IL-1beta, IL-6, IL-8, IL-10, IL-12, and TNFalpha) were measured by ELISA. Group of patients with acute pancreatitis of toxic etiology included 11 subjects: patients from main group received combined therapy supplemented with NSAID from the oxicam class--LX; patients who received only standard basic treatment formed comparison group. RESULTS: It was found that in vitro LX inhibits production of both proinflammatory and anti-inflammatory cytokines by PBMC of healthy subjects mediated by ligands of TLR1/2 and TLR4. Maximal inhibitory effect of LX was observed when cytokine production was induced through TLR1/2. Patients with AP demonstrated increased production of TNFalpha induced by TLR1/2 and TLR4 ligands. CONCLUSION: LX inhibits TLR-mediated production of both proinflammatory (IL-1, IL-6, IL-8, IL-12, TNFalpha) and anti-inflammatory (IL-10) cytokinesby PBMC of healthy subjects in vitro. Treatment with LX in patients with AP results in diminished effector function of TLR1/2 and TLR4 already during 1st day of the illness and normalization of these indices by 6th day.

[Role of innate immunity receptors in development of acute myocardial infarction].

AIM: To assess influence of toll-like receptors (TLR) ligands on the production of proinflammatory cytokine (TNFalpha) by peripheral blood mononuclear cells in patients with acute myocardial infarction (AMI). MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMC) obtained from 13 patients with AMI on 1st and 14th day and from 17 healthy donors were stimulated by peptidoglycan, poly(I:C), lypopolysacchide, zimozan, flagellin and CpG oligodeoxynucleotides, which are ligands of TLR1/2, TLR3, TLR4, TLR2/6, TLR5 and TLR9 respectively. Spontaneous and induced by ligands production of TNFalpha was evaluated in supernatants of PBMC. RESULTS: Increased spontaneous production of TNFalpha by PBMC in patients with AIM was revealed on 1st day of the disease. Ligands of TLR2/6 and TLR4 demonstrated marked stimulatory effect on the production of TNFalpha by PBMC in patients with AMI compared with group of healthy subjects. CONCLUSION: Increased production of TNFalpha by PBMC in patients with AMI indicates the activation of TLR2 and TLR4 on PBMC. Hyperactivation of TLRs during acute pathologic conditions results in excessive production of proinflammatory cytokines, specifically TNFalpha, and can induce damage of cells and tissues involved in acute pathologic process.

[Effect of toll-like receptors (TLR) ligands on in vitro synthesis of proinflammatory cytokines by peripheral blood mononuclear cells from healthy persons and from patients with common variable immunodeficiency].

Production of citokines of tumor-necrosis factor (TNF)-alpha and interferon (IFN)-alpha by peripheral blood mononuclear cells (PBMC) from healthy men and from patients with common variable immunodeficiency (CVID) after stimulation with zymosan, lypopolysaccharides, flagellin and CpG, which are ligands of TLR 2/6, TLR 4, TLR 5, TLR 9 respectively, was studied in vitro. In healthy men production of TNF-alpha varied between individuals, whereas synthesis of IFN-alpha was similar. Spontaneous production of TNF-alpha by PBMC in patients with CVID was increased and accompanied by decrease in TNF-alpha production stimulated by each analyzed ligands except CpG. Observed changes in TLR-dependent TNF-alpha production can play important role in pathogenesis of CVID.

Analysis of toll-like receptor-dependent production of proinflammatory cytokines in vitro by human peripheral blood mononuclears of donors and patients with primary immunodeficiency.

The production of TNF-alpha and IFN-alpha cytokines by peripheral blood mononuclears in response to stimulation by TLR2/6, TLR4, TLR5, TLR9 ligands (zymosan, LPS, flagellin, and CpG-oligodeoxynucleotide, respectively) was studied in donors and patients with common variable immunodeficiency. Individual characteristics of TNF-alpha production by mononuclears were revealed in donors. Reduced stimulated production of TNF-alpha in response to stimulation with TLR4 and TLR5 ligands in vitro was detected in patients with common variable immunodeficiency.

[Congenital components of immunity: Toll-like receptors in the normal state and in immunopathology].

This review deals with rapidly accumulating information on a highly important components of the congenital immune system: Toll-like receptors playing a leading role in the recognition of microbial patterns. The data on the main structural and functional features of Toll-like receptors and their distribution in the body are summarized. The main signal paths are characterized and the key molecules which take part in the transduction are pointed out. Special attention is paid to the activating action of lipopolysaccharides through TLR4. Pathological processes developing as the result of damages in the structure and function of Toll-like receptors in humans and experimental animals are determined.

[The generation of active forms of oxygen by mouse macrophages: the effect of the genotype, immunocytokines and infection]. / Generatsiia aktivnykh form kisloroda makrofagami myshei: vliianie genotipa, immunotsitokinov i infitsirovaniia.

In this work the effect of the natural complex of cytokines on the generation of active forms of oxygen (AFO) by mouse peritoneal macrophage in different in vivo and in vitro systems were studied. Intact mice of various strains were found to have differences in the AFO production of their macrophages: BALB/c (H-2d) > CBA (H-2k) > C57BL/6 (H-2b), as well as in the degree of the prestimulation of the macrophage oxygen metabolism with the complex of cytokines BALB/c > CBA. The in vivo and in vitro infection of macrophages of mice, oppositely reacting to Leishmania donovani, led to the gradual suppression of the capacity of macrophages for AFO generation. The preliminary introduction of the complex of immunocytokines into mice before their infection led to a decrease in AFO generation by macrophages of resistant strains CBA and C57BL/6 and its enhancement by macrophages of the sensitive strain BALB/c.