Platelets are the smallest blood cells, and yet their total volume and surface area exceed those of all types of leukocytes combined. Platelets are produced by the bone marrow megakaryocytes and megakaryocytes in the lung microvessels. Approximately 50% of all platelets are produced in the lungs, which makes it possible to characterize them as the main site for the production of platelets. In small circuit of blood circulation, there are approximately 30% more platelets than in large circuit. This “excess” of platelets is necessary for the stabilization of the endothelial barrier of the lung vessels regulated by the platelet mediator sphingosine-1-phosphate, a regulator of tight junctions of endothelial cells. The circulating platelets have an amazing ability to “bud” new pro- and pre-platelets, giving rise to new platelets. The removal of platelets from circulation proceeds via their phagocytosis by spleen macrophages (if platelets are covered with IgG or are bound to immune complexes), or Kupffer liver cells and hepatocytes (if platelets have incomplete glycans or desialated proteins). In homeostatic conditions, most of the platelets are removed in liver. Platelet clearance in bacterial infections and sepsis is accelerated because of the activity of bacterial sialidases. Recognition of desialized platelet structures is carried out by the liver cells through the Asgr receptor. Despite DNA absence, the platelets are able to synthesize proteins at mRNAs that are present in majority of platelets. Activation of platelets leads to aggregation and exocytosis of the granule contents, and production of immunomodulating molecules. However, activation of platelets may be incomplete and has various consequences. In a non-classical activation model, platelets can release microparticles that contain about 600 different proteins. About 75% of microparticles in the blood of healthy donors are derived from platelets. Like as immune system cells, platelets are activated by numerous endogenous ligands (alarms), including ADP and ATP, which bind to purinergic receptors P2Y1, P2Y12 and
P2X1. Platelets accumulate and retain 99% of the serotonin stored in the body. The platelets contribute to induction of inflammation by releasing proinflammatory cytokines, chemokines, and lipid mediators. In addition, platelets are the source of enzymes that accomplish the capacities of neutrophils and endothelium for production of anti-inflammatory lipid mediators that contribute to tissue repair following acute phase of inflammation.

Tumor necrosis factor alpha (generally known as cachectin) was noted as a hallmark since the time of its discovery and still retains its position as one of the most intensely studied molecules in the field of biomedicine. TNFα is considered a prototype molecule of the family which, on the one hand, plays an important role in regulating normal differentiation, growth and metabolism of various cells, and on the other hand, acts as mediator of inflammatory processes in various human diseases. Intensive interdisciplinary studies of TNFα biological role and therapeutic use have led to understanding of its polyfunctional effects and establishment of its leading role in immune pathogenesis of diseases, which was the basis for the development of “anticytokine” therapy with highly specific monoclonal antibodies. We aimed for a search of literature in the field of studying TNFα structure and properties, its role under normal conditions, in different diseases and therapy. The literature studies were performed using scientific searching systems and databases. The results are as follows: the role of tumor necrosis factor-α (TNFα) in human body in normal and pathological state was considered. Possible TNF-inducible signaling pathways, as well as existing inhibitors based on monoclonal antibodies for the treatment of rheumatoid arthritis and other autoimmune diseases are  described. In conclusion, intensive interdisciplinary studies of TNFα biological functions in normal state and diseases are still relevant today, despite huge knowledge gained on this subject.
Undisputable progress in understanding the functional multiplicity of TNFα as a participant in regulation of various physiological and pathological processes in the human body has promoted development of biological agents, i.e., the tumor necrosis factor inhibitors based on monoclonal antibodies, which proved to be highly effective drugs for treatment of rheumatoid arthritis and other  diseases, thus being effective and safe biopharmaceuticals.

The transmembrane CD200 glycoprotein belongs to the immunoglobulin family and it is widely represented on a variety of cell types, while its structurally similar CD200R receptor is expressed, mainly, on myeloid and lymphoid cells. An immunomodulatory role of CD200 and CD200R interaction is to activate the intracellular inhibitory cascade of reactions, leading to suppression of effector immune cells and attenuation of the inflammatory process. Thus, the CD200R activation stimulates the differentiation of naive T cells to regulatory T cells, increasing the indolamine 2,3-dioxygenase activity, and enhances the synthesis of IL-10 and TGF-β cytokines, contributing to development of a Th2-dependent anti-inflammatory environment. These immune regulatory events provide the development of immune tolerance and are required for controlling the development of autoimmune processes, hypersensitivity, engraftment of transplanted organs and tissues, as well as protecting the fetus from spontaneous abortion. Tolerogenic potential of interaction between CD200 and CD200R molecules can be effectively used for treatment of various diseases (e.g., Alzheimer’s, rheumatoid arthritis, allergies). In this review, we will address the role of CD200/CD200R interactions in stimulating the post-transplant engraftment and protecting a fetus from spontaneous abortion. Many in vivo and in vitro studies have suggested a key role of CD200/CD200R interaction in immune maintenance of both processes. 

Monocytes play a key role in the development of immune response in bacterial infection, because of their phagocytic, antigen-presenting and secretory functions. There are three subpopulations of monocytes: “classical” CD14+CD16-, “intermediate” CD14+CD16+, and “nonclassical” CD14+dimCD16+. These monocyte subtypes have different phenotypes and functions. The ratio of appropriate subpopulations varies with development of the antibacterial response. The aim of the present research was to study phenotypes of the monocyte subpopulations in the patients with sepsis, and changes in the monocyte subpopulation ratio, depending on the presence of bacteria in circulating blood of the patients, as well as to estimate contribution of the monocyte subpopulations to the cytokine production. We observed 16 patients with sepsis (10 men and 6 women; mean age, 58±14 years, SOFA 9.4±2.1; a total of 44 blood samples) examined in dynamics. The control group included healthy adults (n = 23, 12 men and 11 women; mean age, 51±13 years). Laboratory studies included bacteriological cultures, determination of absolute and relative numbers of subpopulations of classical, intermediate and non-classical monocytes and their expression of HLA-DR and CD64, determination of IL-6, TNFα, IL-1β, IL-10 concentration in blood serum. Absolute number of monocytes was increased in the sepsis patients, the ratio of classical monocytes was also increased, like as relative and absolute numbers of  intermediate cells. Meanwhile, the subpopulation of non-classical monocytes did not change significantly. The monocyte subpopulation ratio depended on the presence of bacteria in blood, i.e., a higher proportion of intermediate cells was observed in the samples positive for bacteria in blood cultures. The ratio of subpopulations was restored after elimination of bacteria from the circulation. The expression density of LPS receptor (CD14), IgG receptors (CD16 and  CD64) was found to be increased, especially in the subpopulations of intermediate and nonclassical monocytes. In all subpopulations of monocytes, expression of HLA-DR is reduced, most notably in classical monocytes, least in intermediate cells. There was a significant increase in serum levels of IL-6, IL-1β, TNFα and IL-10 cytokines. Direct correlation between the absolute number of classical monocytes and IL-6 concentration was revealed, as well as intensity of multiple organ dysfunction. Increase in absolute amount of classical monocytes and IL-6 concentration might serve as an indirect criterion for evaluation of endothelial activation, an active producer of IL-6 and myeloid cell growth factors. A direct correlation between the percentage of CD14+CD16+ cells and IL-10 concentration in blood serum indicates to an important role of intermediate monocytes in IL-10 production. IL-10 suppresses the antigen-presenting function of intermediate cells, namely, expression of HLA-DR molecules, as suggested by inverse correlation between the IL-10 concentration and HLA-DR expression density on CD14+CD16+ cells. We have also determined an inverse correlation between the degree of multi-organ dysfunction  and relative amount of HLA-DR+ monocytes. The larger was a classical monocyte subpopulation, the more noticeable was a decrease of this index. The studies in ratios of monocyte subpopulations help to understand the mechanisms of antibacterial protection in sepsis. Monitoring of classical monocyte numbers and serum concentrations of IL-6 is necessary for a comprehensive assessment of inflammatory response in sepsis. Determination of HLADR expression on monocytes allows us to evaluate the intensity of immune suppression in critically ill patients. 

The study was carried out to assess the prospects for the combined use of leukocyte and cytokine indices in the bile duct obstruction of different genesis.
We have performed a study of the patients with mechanical jaundice, either due to tumor (14 persons), or non-tumor genesis (16 persons). Concentrations of cytokines (IL-1, IL-4, IL-8, TNFα) were determined in blood serum before surgical intervention. The total activity of endogenous inflammatory mediators in the serum of patients was estimated as an index of inflammatory activity (IVA), which was calculated using the following formula: IVA = (IL-1 + TNFα)/IL-4. In addition, a number of leukocyte indices were calculated. The cellular component in the index estimation makes it possible to clarify the intensity of the inflammatory reaction, and to suggest the direction of the functional changes among the  immunocompetent cells.
It was shown that IVA in presence of malignancy is significantly higher than in benign conditions. Overactivation of immune response in tumors may be caused by a significant predominance of pro-nflammatory cytokines. Moreover, in cases of malignant tumor growth, both with or without cholangitis, the index scores of humoral and cellular components let us suggest the some problems with regeneration, due to pro-inflammatory domination of immune response, which does not ensure activation of the corresponding reactions. It has been established that the leukocyte intoxication index (LII) is increased,in patients with a combination of malignant disorder and cholangitis, and the immunoreactivity index (MDI) is lowered. In absence of cholangitis, the opposite pattern is observed, when LII is slightly lowered and MDI is elevated. Correlation analysis for the patients with cholangitis revealed a high association of cytokine imbalance and endogenous intoxication. A complex of cytokine and leukocyte indices is promising, when each of them reflects a certain part of the process, and their combination gives an integral picture of the pathology and allows to predict development of the disease. Such an approach, seems to increase the diagnostic significance of these immunological indices, primarily, the cytokine production, and may be used when predicting currence of complications in the surgical treatment of the disease.

Diabetes mellitus type 2 is one of the most important non-infectious diseases in the modern world, being an important risk factor of cardiovascular disorders. Changes in left ventricular myocardial diastolic function are observed in diabetic patients independently from other comorbidities. Etiology of the heart failure during diabetes mellitus type 2 is multifactorial, exhibiting cellular,  molecular and metabolic aspects. However, its pathophysiological mechanisms are not completely understood. The aim of this study was to evaluate numbers of inflammatory T lymphocytes, i.e., T helper type 1 (Th1) and T helper type 17 (Th17) cells, and FoxP3+T regulatory lymphocytes, depending on the functional state of the heart assessed by two-dimensional echocardiography in patients with arterial hypertension and diabetes mellitus type 2. A total of twenty-five patients with a combination of arterial hypertension and diabetes mellitus type 2, and 14 patients with arterial hypertension without carbohydrate disturbances were recruited to a cross-ectional case-control study. All the patients underwent echocardiography with transthoracic access at the M-mode, B-mode and Doppler mode of imaging. We evaluated numbers of Th1 and Th17 lymphocytes by intracellular production of IL-17 and IFNγ by CD4+ lymphocytes, respectively. The numbers of FoxP3+T regulatory lymphocytes were estimated by expression of CD25 and FoxP3 transcription factor. A flow cytometry approach was used in both cases. We revealed some correlations between the numbers of Th17 lymphocytes, FoxP3+T regulatory lymphocytes and functional parameters of myocardium in patients with diabetes mellitus type 2, which were absent in patients
without carbohydrate impairments. The numbers of FoxP3+T egulatory lymphocytes, Treg/Th17 lymphocyte ratio, and mean fluorescence intensity of IL-17 for Th17 cells was lower in patients with diabetes mellitus and diastolic dysfunction compared to the patients with diabetes free of diastolic dysfunction. Association of diastolic dysfunction with diabetes mellitus type 2 was accompanied by increase of IFNγ+Th1 lymphocyte numbers and concentrations of IL-10, IFNγ and TNFα in serum as compared to the patients with diastolic dysfunction in the absence of carbohydrate metabolism disturbances. The diabetic patients with diastolic dysfunction were characterized by hyperinsulinemia, hyperglycemia, higher index of insulin resistance, increase of waist circumference and visceral adiposity index when compared to the patients with diastolic dysfunction without diabetes. Visceral obesity and decrease of insulin sensitivity may be regarded as pathogenetically significant factors for the development of immune regulatory imbalance and diastolic dysfunction in the patients with diabetes mellitus type 2.

Modern studies have shown a high plasticity and phenotypic diversity of neutrophilic granulocytes (NG) provided by different receptors, which are diagnostic markers for the functional capacity of the cell in the course of their activities. We investigated NG from peripheral  blood, obtained from healthy people of both sexes aged from 26 to 66 years. Evaluation of the neutrophil membrane receptor expression was carried out by flow cytometry. The relative amount of neutrophilic granulocytes expressing membrane CD62L, CD63, CD66d receptors and the intensity of their expression were  determined according to their fluorescence intensities. The surface NG membrane receptors, i.e., CD62L, CD63, CD66d were studied upon the in vitro experimental influence of the following bacterial peptides: N-formyl-methionyl-leucyl-phenylalanine (FMLP, model 1); glucosaminylmuramyldipeptide (GMDP, model 2), and simultaneous incubation of NG blood with fMLP and GMDP (model 3). The in vitro treatment with fMLP in the in vitro model was used to transform the NG phenotype of conventionally healthy subjects, expressing CD62, CD63, CD66d molecules. The treatment caused a significantly decrease in both CD62L and the CD62L expression in relative amounts of neutrophilic granulocytes with a parallel increase of CD63 expression density. The effect of GMDP on the NG phenotype of conditionally healthy subjects did not change the amount of CD62L+NG and CD63+NG, and did not affect CD62L and CD63 expression density on the surface of NG. However, the amount of CD66d+NG was significantly increased with the unchanged expression of CD66d molecules. GMDP introduced together with the bacterial fMLP peptide was shown to neutralize some features of the NG phenotype transformation caused by fMLP, i.e., the amount of CD62L+ NG was restored by 22 % and the CD62L expression density increased significantly. At the same time, GMDP did not correct the negative effect of fMLP upon the number of CD63+NG and CD66d+NG, and on the CD63 and CD66d expression. Simultaneous addition of fMLP  and GMDP did significantly increase the amount of CD66d+NG and expression density of CD63 molecules on the CD63+NG membrane as compared to intact NG of conditionally healthy subjects. The obtained data are important in order to justify some new immunotherapeutic strategies aimed at correction of the negatively transformed NG phenotype, which accompanies some infectious and inflammatory diseases of bacterial etiology with atypical clinical course. 

Chronic obstructive pulmonary disease (COPD) is a global public health problem. Studies in immunological features and their correlations with clinical course of COPD are of importance. The aim of this study was to elucidate clinical and immunological features in COPD of different severity grade, concerning Th1- and Тh17-dependent types of immune response.
The study included 132 COPD patients and 32 healthy individuals. According to clinical and functional patterns, the patients with COPD were divided into 3 groups, i.e., 36 cases (28%) of mild severity; 62 individuals (48%), of moderate severity, and 30 patients (23%) of severe clinical grade. We have performed both clinical and immunological evaluation of the patients. The Th1- and Th17-specific lymphocyte subpopulations were assessed according to the serum levels of cytokines, i.e. tumor necrosis factor (TNFα), IL-4, IL-10, IL-17A, IL-21, IFNγ, as well as transforming growth factor β1 (TGF-β1). We have also determined expression of IL-6R receptor (CD126+) on mature T lymphocytes (CD3+) and T helper cells (CD4+) from peripheral blood. We have obtained the following results: the patients with mild-grade COPD exhibited three different T cell phenotypes were determined, with a prevalence of Th1-dependent immune response. The IL-6R were mostly expressed on CD3+CD126+ cells for the Th1/Th17 phenotype, and CD4+CD126+ cells in cases of Th17-dependent type immune response. In patients with COPD of moderate severity, the Th1, Th17, or Th1/Th17 types of immune response was revealed at similar rates. The level of IL-6R expression on mature T lymphocytes and T-helper cells increased to the greatest extent in cases of Th17-dependent immune response. In severe COPD patients, we have found a dominance of Th17 and Th1/Th17 type immune response. The levels of IL-6R expression
were increased in Th17- and Th1/Th17-dependent types of immune response, the most significant increase was observed for CD4+ cells in Th17 phenotype. Clinical features of COPD proved to be associated with the phenotypes of immune response. These results allow of specifying the inflammatory phenotype, predicting the course of chronic disease, and selecting appropriate therapy.

Our aim was to study the effect of cycloferon on in vitro susceptibility of peripheral blood leukocytes to interferon-α2 in children with infectious mononucleosis in acute phase caused by Epstein-Barr virus (EBV).

 Materials and methods were as follows: the study included 23 children 4 to 6 years of age admitted in acute period of infectious mononucleosis. The control group consisted of 15 healthy children of the same age group. In vitro susceptibility of peripheral blood leukocytes to interferon-α2 was determined by the method of L.M. Kurtasova (2007).
Four types of in vitro response of peripheral blood leukocytes to interferon-α2 upon induction by cycloferon have been revealed in both healthy children and patients with EBV-positive infectious mononucleosis. Differences in occurrence rates of these 4 types of reactions have been found among patients with EBVinfection, in comparison with the group of healthy children. There was no regular dose-dependent changes in susceptibility of peripheral blood  leukocytes to interferon-α2 in vitro after cycloferon induction in the group of children with EBV-infection. depending on the dose, which have been revealed in healthy children.

Atypical hemolytic uremic syndrom (aHUS) is a rare severe life-threatening form of thrombotic microangiopathy. aHUS is thought to be primarily mediated by dysfunctional complement regulation, due to mutations or genetic rearrangement of the complement components, or regulatory factors, as well as autoantibody production to the complement factors. These alterations promote uncontrolled complement activation by the alternative pathway on the surface of endothelial cells, followed by endothelial dysfunction, microthrombosis and organ damage, especially, renal pathology. Many studies showed that the biomarkers of endothelial activation/dysfunction including intercellular adhesion molecule type 1 (ICAM-1) and vascular cell adhesion molecule type 1 (VCAM-1) were associated with severe disease and could predict complications and clinical outcomes in different disorders. Increase of these biomarkers was observed in aHUS as well. Until recently, aHUS resulted in unfavorable outcomes, with high death rates in acute phase, and up to 2/3 patients progressed to the end-stage renal failure. Understanding the role of complement dysregulation in aHUS pathogenesis has led to major changes in therapeutic approaches. Eculizumab (a humanized anti-C5 monoclonal antibody) inhibits the terminal complement pathway. This drug has revolutionized treatment and improved prognosis in aHUS. Those patients who received eculizumab have shown sharp decreae in the C3 levels. However, the questions concerning duration of targeted therapy in
remission still remains unclear.
The aim of the present study was to evaluate serum С3, sICAM-1, sVCAM-1 levels in the children with aHUS remission supported by eculizumab maintenance treatment, or without it. Serum C3, sICAM-1 and sVCAM-1 levels were determined in 25 children with aHUS (14 treated with eculizumab and 15, without eculizumab). A control group included 17 children with a history of typical HUS. Serum levels of C3, sICAM-1 and sVCAM-1 were within normal age ranges in all the groups. The children treated with eculizumab showed decreased C3 levels (99±20 mg/l vs 112±15 mg/l, and 123±40 mg/l, respectively), and increased sICAM-1 levels (483±103 ng/ ml vs 343±50 ng/ml and 401±91 ng/ml, respectively) compared to other groups (p < 0.05). No differences in sVCAM-1 levels were revealed in the groups. Hence, no signs of subclinical complement activation or endothelial disfunction were revealed in the group free of eculizumab therapy. Normal C3, sICAM-1 and sVCAM-1 levels in blood indicate that normal endothelial state could be restored in aHUS, and this condition is maintained after discontinuation of the targeted therapy. Our results suggest that C3, sICAM-1 and sVCAM-1 monitoring may be useful for further management of these patients and for prediction of relapses.

The aim of this prospective randomized clinical study was to investigate the role of preoperative carbohydrate admnistration in surgery-induced metabolic, immune and inflammatory reactions after thoracoabdominal operations. At the Surgical department I (B.V. Petrovsky National Research Center of Surgery), we investigated a modulatory role of carbohydrate preload upon surgical stress observed after major thoracoabdominal operations (thoracoscopic and open esophagectomy, retrosternal colonic esophagoplasty) followed by the enhanced recovery protocol. The study was performed in 2014-2017, it included 30 patients, divided into 2 groups. Group A patients (n = 16) received carbohydrates preload (12.5% maltodextrin solution per os or enterally). In patients with dysphagia, the 12.5% dextrose solution was used intravenously in equal volumes. Group B patients didn’t receive any additional preload with carbohydrates. The groups were age- and gendermatched, similar for disease and surgery types. Glucose and insulin levels (with HOMA insulin resistance index, HOMA-IR) were measured before surgery and on day +1, interleukin levels (IL-6, IL-10, IL-8) and index IL-8/IL-10 were assessed before surgery, and on days +1 and +5 after surgery. Cell-mediated immunity was investigated before surgery and on day +5.
The stress-induced hyperglycemia (> 7.8 mmol/L) was detected more frequently in group B (50%), than in group A (6%), p = 0.012. Insulin resistance measured by HOMA-IR in group B was detected in 71% of patients and in 25% patients of group A only, p = 0.027. Individual analysis of immune response demonstrated that a trend for immune recovery was detected by the day +5 post-op in the group A. Postoperative levels of IL-6 and IL-10 were lower on day +1 and +5 in group A. Morbidity rates and the terms of hospitalization were similar in both groups. Local postsurgical infections in group A were developed in 6% of the patients vs 35.6% in group B (p = 0.072).
In conclusion, a complex study of surgical stress, i.e., metabolic, immune and inflammatory reactions after esophageal surgery has shown that the carbohydrate preload decreased the incidence of postoperative insulin resistance and stress-induced hyperglycemia, being accompanied by lower release of proinflammatory cytokines and provides positive effects upon the patient’s immune system.

Our aim was to study the influence of IL4-589C>T gene polymorphism on IL-4 expression in patients, living in Zabaikalsky Region with a diagnosis of chronic traumatic osteomyelitis developing after fractures of long bones of extremities.
The study included 132 patients with fractures of long bones at the age of 20 to 40 years old. The first group consisted of 83 patients with uncomplicated course of fractures; the second group (n = 49) included patients with chronic traumatic osteomyelitis healed by primary wound closure, however, developing chronic traumatic osteomyelitis over late postoperative period. The control group consisted of 100 practically healthy men and women of the same age group. These groups were homogeneous and similar by age, sex, origin and localization of fractures. Presence of acute or chronic comorbidities was the exclusion criterion. We applied clinical examination, instrumental tests (X-ray studies), laboratory methods (microbiological, immunological, ELISA for IL-4 measurement). IL-4 point mutation at position 589 (C>T) was tested by PCR; the DNA for this analysis was extracted from peripheral blood. The genetic studies were performed when patients entered the hospital. IL-4 contents, clinical and instrumental indicators were detected on 1, 2, 10 and 90 days after fracture. 

Results of the study were as follows: the frequency of the C allele and the homozygous genotype of the IL4- 589C>T gene in the patients with complicated clinical course was decreased 1.7- and 3.6-fold compared with the comparison clinical group, respectively. Frequency of T allele, by contrast, increased 3.7-fold, whereas heterozygous and mutant genotypes were changed 2.5 and 15 times against the comparison group. The persons from control group with IL4-589C/C genotype, showed a 1.4- and 2.5-fld increased IL-4 concentration as compared with the carriers of the -589C/T and -589T/T genotypes, respectively, and in -589C C/T genotype, 1.7-fold compared to -589Т/T genotype. A similar trend of IL-4 expression, depending on the IL4 genotype, was recorded in the group with both uncomplicated and complicated course of fractures. In conclusion, the patients with developing traumatic osteomyelitis showed ncreased frequency of the IL4 -589T allele (3.7-fold as compared with uncomplicated group). Meanwhile, higher frequency of the -589C/T -589T/T genotypes of the IL4 gene were registered. Genotype -589T/T of the IL4 gene is associated with lower IL-4  expression.

The prolonged impact of industrial vibration on workers leads to the development of a vibrational disease (VB), which occupies a leading position in the structure of occupational pathology. VB from the impact of local vibration is a chronic occupational disease characterized by a predominant lesion of the nervous, vascular system and musculoskeletal system of the upper and lower extremities. One of the real ways to reduce the incidence is the early detection of the negative impact of vibration on the body of workers. In this regard, cytokines and heat shock proteins (HSP70) can be early and sensitive indicators that reflect the severity of health disorders from exposure to vibration. The aim of the study was to study changes in the content of pro- and antiinflammatory cytokines, extracellular HSP70 and their relationship in patients with VB. In the immunological study included 43 male patients with a diagnosis of VB. The criteria for inclusion in this group were: a verified diagnosis, written informed consent to participate in the study, the harmful effects of local vibration in the workplace. According to the data of hygienic control, the working conditions of workers in dangerous occupations by vibration belong to the 4 (dangerous) class due to intensive local vibration. The content of cytokines: IL-1β, IL-8, IL-10, TNFα and HSP70 in the serum of patients was determined by enzyme immunoassay. The blood sampling for the study was taken from patients only once on admission to the hospital before the treatment. Statistical processing of data was carried out with the help of packages of application programs Statistica for Windows 6.0 and Microsoft Excel. It was established that the development of VB disease is accompanied by imbalance of the cytokine profile, characterized by a decrease in the levels of IL-1β, IL-10 and an increase in IL-8. The inhibition of production of IL-1β and IL-10 is a consequence of the chronic process that has developed in the body of the patients examined. And the revealed decrease in extracellular concentration of HSP70 in comparison with practically healthy people can be caused by the accumulation of it inside the cell or on its surface. Correlation analysis revealed the relationship between a decrease in HSP70 and an increase in IL-1β, and a decrease in IL-10 levels. Synthesis of HSP is an intracellular defense mechanism that prevents cell damage by activating the synthesis and secretion of pro-inflammatory cytokines. The obtained relationships between cytokines and HSP70 testify to the involvement of HSP70 in immunoinflammatory processes in VB. The revealed changes contribute to the chronic inflammatory process and justify the progressive course of the VB.

Evaluation of the immune system parameters can be used in order to assess capacity to adapt under conditions of increased external chemical load, including exposure to metals, which can exert either activating and inhibitory effects upon immune regulation parameters. The aim of this work was the analysis of immunoregulatory markers in a children’s population who consumed water with high strontium content (a sample from the Perm region). We carried out immunological evaluation of the children aged 7 to 12 years, living at a territory with a high strontium content in the drinking water. The comparison group included children from the conventionally clean region. We studied differential changes in cellular immunity (phagocytosis rates), humoral factors of immune defense (serum immunoglobulins), development of specific sensitization for strontium, as well as the processes of apoptosis triggering and regulation. A 3.68-fold increase in strontium levels was shown in fresh water within observation area, and the average  blood strontium content in the children of appropriate observation group was 1.55-fold higher than in children of the comparison group. At the same time, 1.2-fold increase in phagocytic activity determined as phagocytic number and phagocytic index was found, as compared to the control group. In 80% of the subjects, a reduction in serum IgG level was observed when compared to physiological norm, as well as a significant decrease in IgG and enhance in IgM production against the levels found in the comparison group. Wehave also shown an enhanced total sensitization in 55.0% of the observation group as shown by the total IgE test compared with normal age ranges, as well as excessive specific sensitization to strontium by 2.49 times, according to the IgG criterion. Disturbance in apoptosis triggering was associated with decreased number of CD95+ lymphocytes and TNFR1+ cells (2.8-fold compared to reference values), shifted balance in apoptogenic proteins, an average of 2.6-fold decrease in Bcl-2 expression, a 2.8-fold reduction of the p53 transcription factor expression relative to the reference interval. Thus, we have shown an ability of strontium excess in drinking water to influence the most important indices of immune regulation in pediatric population. These changes may serve as indices of populational health status under of external strontium exposure.

In the present study, we have examined association between different polymorphic variants of metalloproteinases genes and clinical manifestations of bronchial asthma in children. We observed 103 patients including 42 children with an established diagnosis of asthma. Moreover, 61 persons were examined in the control group. All patients underwent genetic testing by allele-specific polymerase chain reaction. In particular, 320A>C polymorphic locus of ММР20 gene; Val275Ala ММР20, and -8202A>G gene ММР9 were analyzed.
We have found that 30 patients (71.4% of total) had bronchial asthma of mild severity, 9 children (21.4%) exhibited moderate degree, and 3 patients (7%) had severe-grade disease. Homozygous C/C variant of the polymorphic ММР20 gene, 320A>C heterozygous variant of the ММР20 Val275Ala polymorphism, and heterozygous locus of -8202A>G ММР9 gene were found to be most frequent among the children with asthma. Generally, we have observed that the frequencies of the studied alleles and genotypes did not significantly differ berween the asthma patients and children from the control group (p < 0.05). However, in patients with GGgenotype of -8202A>G ММР9 polymorphism combined with homozygosity for the C allele of ММР20 320A>C, a more severe disease was observed, being combined with polyvalent sensitization and high total IgE levels in blood serum.
In conclusion, frequencies of alleles and genotypes among patients with asthma did not show any statistically significant differences from the group of healthy children. The patients homozygous for G allele of ММР9 -8202A>G polymorphism gene and for the C allele ММР20 gene (320A>C) seem to be predisposed for a more severe clinical course of the disease. 

Combination of bronchial asthma (BA) and obesity is a difficult-to-control phenotype. Studies of inflammatory process with respect to severity of the disease are important for understanding the potential influence of obesity on the BA clinical course. The objective of this study was to determine cytokine profile in patients with mild BA combined with obesity. The study involved fifty-three patients with partially controlled mild BA. The patients were recruited as volunteers and signed an informed consent. The first observation group consisted of 27 asthma patients with normal body weight, the second observation group consisted of 26 patients with BA combined with obesity. A control group included 25 healthy volunteers. All the patients underwent clinical and laboratory examination in accordance with clinical standards for BA and obesity. The levels of TNFα, IL-2, IL-4, IL-6, IL-10 were evaluated in blood serum by means of flow cytometry. The ratios of proand anti-inflammatory cytokines (TNFα/IL-4, TNFα/IL-10, IL-6/IL-4, IL-6/IL-10) were calculated. Asthma patients with obesity (the 2nd group) had elevated levels of IL-2 over control group and group 1, by 38% and 44% respectively(p < 0.05). The concentration of proinflammatory cytokines TNFα and IL-6 was significanty increased in the both patient groups. Mean TNFα level was increased 2.5 times (p < 0.05), and IL-6 levels were increased by 30% (p < 0.05) in the 1st group as compared to the controls. TNFα and IL-6 concentrations showed a 3-fold increase over control values (p < 0.05) in the 2nd group. The level of antiinflammatory cytokine IL-4 was increased in patients with BA, independently of body mass. It should be noted that the concentration of this cytokine in obese patients was higher by 29% than in patients with normal body weight. IL-10 levels in patients from the 2nd group were reduced more than 2 times than in the 1st group. The patients of the 1st group showed a decrease in the IL-6/IL-10 index, in comparison with control parameters, thus indicative of an imbalance due to the elevation of the anti-inflammatory IL-10 cytokine. Among BA patients with obesity (group 2) the TNFα/IL-10 and IL-6/IL-10 indexes were higher than those of the control group (2.3- and 5.5-fold, respectively) and the group 1 (2.6- and 2.5-fold, respectively). Dynamics of these indexes confirms the systemic nature of inflammation and a predominance of non-atopic  inflammation in asthma patients with obesity. Thus, features of the cytokine profile in BA with obesity consist of a significant increase in pro-inflammatory IL-2, IL-6, TNFα cytokines, and a relative decrease in anti-inflammatory IL- 10 cytokine. The development of BA with obesity, even in mild-severity BA, is accompanied by development of a cytokine disbalance, which is typical for a mixed-type inflammation, with a prevalence of neutrophil inflammation. 

Сomplex intergenic interactions should be taken into account when predicting the risk of adverse course in a pathological process. This presumption is at the heart of evolving multifactorial diseases (ulcerative colitis and irritable bowel syndrome). A single genetic polymorphism seems to be a weak risk factor when predicting the disease evolution and it could not be used as a prognostic model for development of multifactorial pathologies, especially in cases of rare alleles. However, it is well known that the combination of unfavorable alleles of several genes with an additive effect is dangerous. Therefore, identification of such polymorphisms is very important.
Previously we conducted a test panel, in order to evaluate associations of alleles and genotypes of some cytokine genes (IL1β, TNFα, IL1rа, IL10, IL6) with predisposal, or resistance for ulcerative colitis and irritable bowel syndrome in the Russian Chelyabinsk region. Distribution of alleles and genotypes of these genes have been assessed in irritation bowel disease (IBD) and ulcerative colitis (UC). The following methods were used: isolation of DNA samples from whole blood, genotyping of the studied gene polymorphisms with PCR, RFLP. A comparative analysis of intergenic interactions between the cytokine genes IL1β, TNFα, IL1ra, IL10, IL6 in the IBD and UC patients was carried out by the Multifactor Dimensionality Reduction method. The analysis is based on the use of polymorphic loci of IL1β, TNFα, IL1ra, IL10, IL6 combinations chosen for analysis of intergenic interactions, with respect to the risk for IBS and UC predisposition. As a result of this study, a fourlocus model IL1β(+3953)*Т/TNFα(-308)*A/IL10(-1082)*G/IL6(-174)*G was identified for IBS, which was characterized by 90% repeatability and prediction accuracy of 74.5%. This model of gene interactions between the cytokine IL1β, TNFα, IL1ra, IL10, IL6 genes had the greatest prediction potential (p < 0.001) in IBS, whereas the model for UC was not statistically significant. The following types of genetic interactions were established: synergism between the IL1β(+3953)*Т and TNFα(-308)*A loci, whereas IL6(-174)*G и IL10(-1082)*G, TNFα(-308)*A seem to be in antagonistic relationships. The study made it possible to establish that the -174G/C IL-6 polymorphism may play a central role and provide intergenic interactions with SNPs of IL1β, TNFα, IL10 for predisposal to irritable bowel syndrome in Chelyabinsk Region of Russia.

Previous studies reported some associations between class A antibodies specific for benzo[a]pyrene (IgA-Bp), estradiol (IgA-Es) and progesterone (IgA-Pg) and breast cancer (BC) in women like as with lung cancer (LC) in men. It was suggested that IgA-Bp and IgA-Es may stimulate tumor initiation and promotion, whereas IgA-Pg may inhibit the in vivo human carcinogenesis.
The purpose of this study was to identify the suggested associations of such immunological imbalance with BC and LC in postmenopausal women.
The serum A-class antibodies specific to benzo[a]pyrene, estradiol and progesterone (IgA-Bp, IgA-Es, IgA- Pg) were studied in 335 healthy women, 824 breast cancer (BC) patients and 127 cases of lung cancer (LC) by means of non-competitive solid phase immunoassay. The following results were obtained: Increased ratio of IgA-Bp and IgA-Es amounts exceeding the IgA-Pg levels was associated with a higher risk of breast cancer (OR = 2.8 and 2.4 respectively, p < 0.0001), and higher risk of LC (OR = 2.9 and 2.8, respectively, p < 0.0001). Conversely, the OR values decreased to 0.3-0.4 for BC and LC if IgA-Pg levels were higher than IgA-Bp and IgA-Es levels (p < 0.0001). These findings confirm the hypothesis that IgA-Bp and IgA-Es are capable to stimulate, and IgA-Pg, to inhibit the BC and LC occurrence n postmenopausal women. The balance between IgA-Bp and IgA-Es, on the one hand, and IgA-Pg, on the other hand, is much more important than individual contents of these antibodies.
In conclusion, the phenomenon of “immunological interference” is revealed, i.e., the mutual enhancement of IgA-Bp and IgA-Es effects, thus, probably, stimulating the initial and subsequent events of carcinogenesis initiation and promotion, with a weak anticancer effect of IgA-Pg, and by weakening the mutual procarcinogenic effects of IgA-Bp and IgA-Es by the marked effect of IgA-Pg.

Production of antibodies to erythropoietin-stimulating drugs is an important problem of therapy with recombinant human erythropoietin (EPO). It leads to changes in the pharmacokinetic profile and decreased therapeutic efficiency. Upon long-term treatment with EPO preparations, neutralizing antibodies can result in rare cases, thus leading to complete pure red cell aplasia. Hence, detection of antibodies to EPO is an important stage in the assessment of the drug immunogenicity in preclinical and clinical studies, as well as during the treatment with EPO. We have compared colorimetric and chemiluminescent ELISA tests for detection of IgG antibodies to human EPO with 3,3’,5,5’-tetramethylbenzidine – hydrogen peroxide and luminol -hydrogen peroxide detection systems, respectively. Аntibodies to human EPO were determined in blood serum samples of experimental animals, i.e., rabbits and guinea pigs following their immunization with
different doses of pegylated human recombinant EPO-beta subcutaneously or intravenously. The affinitypurified rabbit polyclonal antibodies to human EPO were used as a reference material. The effects of hydrogen peroxide and luminol concentrations upon sensitivity of a chemiluminescent method were also studied. We have shown a 1.5-2-fold increase in sensitivity when using 4-iodophenol for amplification of chemiluminescence. A comparison of the chemiluminescence intensity with time has demonstrated a better stability for the substrate mixture prepared on borate buffer. A decrease in chemiluminescence signal with time was proportional to the decrease in background signal, thus rendering stability of the signal/background ratio for 3 to 30 minutes. Due to optimizing the substrate mixture composition and conditions of chemiluminescence recording, the reached detection limits for colorimetric and chemiluminescent ELISA’s were, respectively, 0.6 ng/ml and 0.08 ng/ml. The measurement range was extended by more than 20 times for chemiluminescent ELISA. The chemiluminescent ELISA for anti-erythropoietin antibody detection showed a 1.9 to 2.6-fold increase in sensitivity for rabbit serum, and 1.8 to 8.9-fold for guinea pigs serum. Good correlation of results was found for quantitative detection of antibodies in rabbit sera using the two methods (R = 0.981). Thus, chemiluminescent ELISA allowed develop a more sensitive detection technique of IgG antibodies to human erythropoietin.