The existing data on regulatory T cells (Tregs) in prostate cancer suggest that these cells may penetrate the prostate gland malignant tissue, suppressing antitumor immune response, thus promoting aggressive clinical course and low survival of the cancer patients. Evaluation of T cell subpopulations from the tumor microenvironment has shown that the number of CD4+Tregs is associated with inferior clinical prognosis. In particular, each additional CD4+Treg cell has been shown to cause a statistically significant increase in prostate cancer mortality by 12%, regardless of other clinical factors. There are several possible explanations for the increased infiltration of prostate cancer tissue with regulatory T cells. Firstly, malignant cells or tumor-associated macrophages are capable of secreting chemokine CCL22, which has an affinity for the CCR4 receptor expressed on Treg cells. Secondly, cytokines secreted by prostate tumors, such as TGF-β, may regulate the FoxP3 expression, thus expanding the Treg population. TGF-β, in turn, is a multifunctional cytokine that promotes survival and proliferation of transformed cells, including prostate epithelium, as evidenced by increased amounts in the patients with metastatic disease.

Response of human natural killer (NK) cell populations (NKP) against tumors in the presence of viruses was evaluated as a quite variable, early adapting for the pathological signals in organism, mobile and selective combination agents. NKP act as a result of co-functioning between the receptor lectins (RL) recognizing glycopatterns (RL as triggers, initiators and basic agents for coupled activities), and Ig-like, cytotoxic and other additional  communicative and effector receptors (superstructural, tuning for achievement of final effector-type NKP constructions required), and their ligands (modulators of final cell surface receptor mosaics). Such NKP created play important role in redistribution of NKP-induced antitumor/ antiviral cytokines in organism. Intercellular communicative potential of NKP also involves other innate and innate-like cells. Such extended communications of NKP provide a prospective and universal resource of human protection. NKP must be under consideration upon development of new maneuvre and relilable prophylactic and immunotherapeutic antitumor/ antiviral systems and vaccine strategies. The ways for the fine tuning (RL—KIR/ NCR/ CD/ their combinations) algorithms of RL-based creation of antitumor/ antiviral NKP are revealed. Key role is given to screening spectrum of patient NKP for development of communicative anticancer/ antiviral strategies. The status of NK compartment will characterize resistance of individuum/ contingent of individuums to viral infections of epidemiological significance, will play important role in anti-epidemic protection of regional population.

The review presents data on the role of glycodelin A (GdA, PP14, α2-PEG, EP15, PEP, AUP, PAEP) in regulating the functions of the immune system in the context of the formation of feto-maternal immune tolerance during pregnancy.

Glycodelin was first isolated and identified in 1976 by D.D. Petrunin. and Tatarinov Yu.S. with colleagues as a new placenta antigen, which was named chorionic α2-microglobulin. Since then, a huge amount of scientific data has been obtained on the structure, properties, and biological effects of this glycoprotein. This protein has four differentially glycosylated isoforms, namely GdA, GdF, GdC, and GdS, which are secreted in different parts of the reproductive system.

The most studied isoform, glycodelin A (GdA), is secreted by the decidual glandular epithelium and accumulates in the amniotic fluid and maternal serum during pregnancy. GdA level is a sign of endometrial fertile function. GdA has diverse biological effects, in particular, it modulates endocrine function and differentiation of trophoblast cells.

The role of GdA in the regulation of the immune system is to inhibit the proliferation of T and B lymphocytes, suppress the cytotoxicity of NK cells, induce apoptosis of activated CD4+ cells, monocytes and NK cells, inhibit the activity of cytotoxic T lymphocytes and suppress the functional activity of macrophages and dendritic cells. In addition, GdA increases the level of regulatory T cells, regulates the Th1/Th2 balance towards Th2, and induces a tolerant phenotype of dendritic cells.

The immunomodulating activity of GdA depends on the degree of its glycosylation, which, in turn, is associated with the preparation obtaining method. Therefore, the review analyzed the features of the immunomodulating effects of the native and recombinant types of glycodelin on the immune system cells.

However, the cumulative effects of GdA on the cells of the immune system make it possible to consider it as one of the main factors shaping the feto-maternal immune tolerance during pregnancy. It is also important to note that clinical studies have revealed a correlation between low levels of circulating GdA and repetitive spontaneous abortions that confirms the importance of this protein in fetoprotection.

In general, it is obvious that GdA has the potential as a medicinal preparation for autoimmune deseases treatment, post-transplant complications and in vitro “reprogramming” of autoreactive T-cell clones for further cellular immunotherapy.

Chemokines are a special family of cytokines whose main function is to control cell migration; they are key players in the innate and adaptive immune responses. Directed chemotaxis of specific leukocyte subpopulations is necessary not only to maintain homeostasis, but also in development of some immunopathological conditions such as cancer, inflammation, infection, allergies and autoimmune disorders. Chemokines are pleiotropic molecules that are involved in physiological and pathophysiological processes. For example, the CXCR3 chemokine receptor is expressed on various cells: activated T and B lymphocytes, natural killers, eosinophils and neutrophils, dendritic cells, fibroblasts, endothelial and epithelial cells. Hence, CXCR3 and its ligands have a wide range of functional activity. CXCR3 ligands are the IFNγ-induced chemokines: CXCL9, CXCL10, CXCL11, and platelet-derived chemokines: CXCL4, CXCL4L1. All the CXCR3 ligands share common angiostatic properties due to lack of the Glu-Leu-Arg (ELR) motif. IFNγ-induced ligands of the CXCR3 are proinflammatory chemokines, they mainly recruit activated T cells and exert an effect on T cell polarization. Due to wide spectrum of biological activity, the ligands of CXCR3 receptor are involved in pathogenesis of various disorders, such as inflammation, infection, cancer, allergies and autoimmune disorders. In this review, we discuss the role of CXCR3 ligands in immunopathogenesis of various diseases, including the results of our studies in chronic hepatitis C, rheumatoid arthritis and pulmonary tuberculosis. Moreover, we have also discussed the potential laboratory diagnostic applicability of the chemokines in various diseases. This review illustrates a universal role of IFNγ-induced chemokines as mediators of immune responses in various diseases. The studies of CXCR3 ligands, their isoforms and receptors, interactions between themselves and with their receptors can provide a significant contribution to our understanding of the chemokine network. Understanding the system of IFNγ-dependent chemokines may have clinical implications, both for diagnostic tasks, and for therapeutic purposes.

Perivascular adipose tissue surrounds most large blood vessels and plays an important role in vascular homeostasis. Recent studies have shown that perivascular adipose tissue effects vasodilation and vasoconstriction, which indicates that the perivascular adipose tissue regulates tone and diameter of the vessels. Adipocytokines and chemokines, secreted from the perivascular adipose tissue, apparently have direct access to the adjacent arterial wall by diffusion, or through vasa vasorum. The present data indicate that perivascular adipose tissue plays an important role in atherosclerosis, hypertension. Recently, some studies were performed that examined the role of perivascular adipose tissue in non-atherosclerotic vascular diseases, such as neointimal formation, aortic aneurysm, arterial stiffness and vasculitis. The present review will discuss a role of perivascular adipose tissue in pathogenesis of atherosclerosis, hypertension and non-atherosclerotic vascular diseases.

Hypoxia and immune reactions are closely interrelated at molecular, cellular and organism levels, and the individuals differ in resistance to oxygen deficiency. Animals with high and low resistance to hypoxia have different adaptive capabilities and predisposition to the development of inflammatory diseases. Data on the individual characteristics of hypoxia resistance in female laboratory animals and humans, and its relationship to immune system reactions in both normal conditions and inflammatory diseases are not available in the literature. It is known, however, that acute infectious and inflammatory diseases develop at lesser rates and are less severe in women and female laboratory animals than in males, which can be explained by higher resistance of females to hypoxia. The aim of our study is to reveal the features of morpho-functional thymus changes, and subpopulation of peripheral blood lymphocytes in systemic inflammatory response induced by LPS administration to female Wistar rats with different resistance to hypoxia. 

Resistance of mature female Wistar rats to hypoxia was determined as a survival period in a ventilated lowpressure chamber simulating high altitude condition (11 500 m). The rats with a lifetime “at high altitude” of > 180 s have been classified as highly resistant to hypoxia, and the animals surviving for < 20 seconds were designated low-resistant. One month after determining the hypoxia resistance, the females were injected intraperitoneally with E. coli O26:B6 lipopolysaccharide (LPS) at a dose of 1.5 mg/kg during the dioestrus phase. The animals were withdrawn from the experiment by i/m Zoletyl injection (15 mg/kg) one day after LPS administration. The relative volume fractions of thymic cortex and medulla were evaluated; the areas of necrosis were determined in the liver, and the number of neutrophils in the interalveolar septa was counted in the lungs. The serum contents of corticosterone, testosterone, TGF-β were determined. A flow cytometry evaluation of the relative and absolute numbers was performed for major subpopulations of lymphocytes in peripheral blood. The number of apoptotically dying cells of the thymus was assessed. For statistical processing of the obtained data, the Statistica 8.0 software was applied, using criteria of multiple comparisons by Kruskal–Wallis and Dann. The differences were considered statistically significant at p < 0.05.

In both high- and low-resistant to hypoxia females, the development of a systemic inflammatory -response was accompanied by a moderately severe thymic involution, apoptosis of thymocytes, an increase in the absolute number of NK, and rise of testosterone and corticosterone contents. LPS injection into low-resistant rats, if compared to females highly resistant to hypoxia, led to more severe manifestations of systemic inflammation, i.e., a pronounced inflammatory reaction in the lungs and a more extensive liver necrotic area accompanied by increased absolute numbers of regulatory T lymphocytes and T helper cells, and more pronounced thymic accidental involution with apoptotic death of thymocytes. Systemic manifestations of inflammation were less pronounced in hypoxia-resistant female rats, which was apparently associated with activation of lymphocyte  migration from the thymus and blood to the inflammation focus, and development of more effective immune response.

Conclusion: immune reactions in the systemic inflammatory response induced by LPS in female Wistar rats depend on individual resistance to hypoxia. These data should be used to develop approaches to personalized therapy of infectious and inflammatory diseases in women.

The immunomodulatory activity of vascular endothelial growth factors (VEGFs) reveals a new role of neoangiogenesis in tumor development. Most of VEGF effects on T cells are mediated through the VEGF-R2 receptors. Placental growth factor (PlGF) belongs to the VEGFs family and is a selective ligand for VEGF-R1. In order to study the role of VEGF-R1-signaling in the regulation of T-cell functions, the effect of PlGF on the proliferation of donor T cell has been investigated. PlGF has been shown to inhibit the proliferation of T-lymphocytes in cultures of anti-CD3-stimulated mononuclear cells in a wide dose range, suppressing the proliferative response of both CD4 + and CD8 + T cells. The suppressive effect of PlGF was mediated through the direct interaction with VEGFR-1 on T-cells that was evidenced by the expression of VEGFR-1 by T-lymphocytes (especially after their activation) and by blocking the suppressive effect of PlGF with neutralizing anti-VEGFR-1 antibodies. Given the increased levels of PlGF in many tumors, this factor may play an important role in immunomodulation during tumor growth, mediating its effect through the VEGFR-1 signaling pathway.

Intensivity of lymphocyte proliferative activity is an important and almost the only indicator of the level of functional activity of these cells, and the immune system in general. The aim of this study was to develop an adequate physiological test to determine the degree of functional activity of lymphoid cells. The choice of IL-1 as an activator of proliferation is justified by the presence of receptors to IL-1 on lymphoid cell membranes, the key role of this cytokine in the initiation of a wide range of biological effects of the immune system to antigen, including participation in intercellular cooperation and distant action on various cells. It was found that the ligand-receptor interaction of IL-1 by type I receptor activates neutral sphingomyelinase, and the sphingomyelin pathway of signal transduction in lymphocytes. The degree of activation of this enzyme depends on the type of stress and correlates with changes of the vector of humoral immune response and proliferative activity of lymphocytes. The level of IL-1 increases, but the activity of neutral sphingomyelinase and lymphoid cell proliferation is significantly reduced after applications of immunosuppressive stress in mice and rats. The change of the degree of lymphocyte proliferation has become a marker of the severity of the pathological process in clinic. A high degree of correlation between the low intensity of peripheral blood lymphocyte proliferation in response to IL-1 action and the unfavorable disease outcome in patients with severe combined trauma and children with purulent meningitis was established. Thus, the degree of lymphocyte proliferation in response to the action of the regulatory signal of IL-1 can be used both for the analysis of the effectiveness of immunomodulators, and as diagnostic and prognostic indicator in clinical practice.

Natural killer (NK) cells are of special interest among a multitude of microvesicle (MV) source cells. NK cells are a lymphocyte subpopulation performing contact cytolysis of virus-infected cells and tumor cells. Each of the NK cell populations has a unique receptor repertoire on its surface and, thus, unique functions. During their contact with a target cell, the most common mechanism of cytolysis is an exocytosis of lytic granules. However, some indirect evidence suggests that MV with CD56 phenotype and leukocyte-derived MV with various phenotypes are present in the peripheral blood plasma.This research is aimed to study the phenotype, composition and cytotoxic activity of microvesicles produced by NK cells. The analysis of receptor expression showed that MV, as well as source cells of the NK-92 cell line, had a similar CD56 molecule expression profile. The expression profile in MV differs from the same in source cells by higher CD119 and CD11b expression and by lower CD18 expression. Culturing of NK-92 cells in the presence of PMA, IL-1β, TNFα, IFNγ resulted in alterations of cell phenotypes and MV. Immunoblots revealed a change of perforin and granzyme B (GrB) in MV. The analysis of the cytotoxic activity of NK-92 cells in a natural killer in vitro assay employing K562 target cells demonstrated that MV obtained from TNFα-activated cells of the NK-92 cell line increased the cytotoxicity of the same TNFα-activated NK-92 cells regarding cytotoxicity levels. This coincides with the previously revealed increased content of GrB in MV obtained from TNFα-activated cells of the NK-92 cell line. To sum up depending on the cytokine NK-92 cells produce MV that differ in their phenotype, composition and activity. Any changes in MV composition can result in changes in their functional activity: in particular, changes can increase the cytotoxic activity of NK cells of the NK-92 cell line. Thus, besides a well-known and proved way for GrB delivery to a target cell, we can suggest an additional way – the transportation of GrB within MV.

The aim of the study was to investigate an interdependence between the phenotype of dendritic cells (DC) differentiated from monocytes and the number of pro-inflammatory monocytes in peripheral blood of patients with kidney cancer (KC). The study involved 28 patients at the age of 40-55 years suffering with KC (Т3N0М0, clear cell type) before surgical treatment. The diagnosis was verified histologically. 31 healthy agematched persons were examined as a control group. Mononuclear cells were isolated from heparinized venous blood by centrifugation in a Histopaque®-1077 density gradient followed by plastic adsorption in RPMI 1640 medium supplied with 10% autologous serum. Immature DCs (iDCs) were generated from blood monocytes by culturing for 5 days with GM-CSF and IFNα. Activation of DCs (mDCs) was induced by incubation with the tumor cell lysate and TNFα, followed by incubation for 48 hours. A tumor fragment was used to prepare the lysate of autologous tumor cells. Phenotyping of blood monocytes and DC at various maturation stages was performed by flow cytometry. The numbers of CD14+CD16+ monocytes in peripheral blood of KC patients were decreased (up to 42% of the total monocyte level) against the control ranges. In this regard, the analysis of the dependence between the phenotype of DCs differentiated from monocytes and the number of pro-inflammatory blood monocytes was carried out by comparing the groups with a high content of pro-inflammatory monocytes in the blood in KC patients (> 42%, near-control range) and low content (resp., < 42%). We have found that the contents of tolerogenic iDC in cell culture are increased in KC patients with low amounts of pro-inflammatory monocytes in blood (< 42%). A relatively increased expression of antigen-presenting and co-stimulatory molecules proved to be the specific feature of iDC phenotype in patients with high contents (> 42%) of proinflammatory monocytes in blood. The phenotype of dendritic cells in KC patients with different content of proinflammatory monocytes during maturation/activation showed more differences. In the patients with low levels of pro-inflammatory monocytes, the cell pool of in vitro maturing DCs was characterized by low level of CD86 and HLA-DR receptor expression, thus reflecting a weak co-stimulating and antigen-presenting activity. In the patients with high levels of pro-inflammatory monocytes in blood, the in vitro activated DCs showed higher level of functional activity using the above markers. The revealed differences in the DC phenotype and interrelations with amounts of blood monocyte subpopulations in KC patients may presume the programmed cell differentiation mechanisms depending on the microenvironment, under pathogenic conditions (i.e., in presence of malignant tumor growth).

Multiple myeloma is the most common form of paraproteinemic hemoblastosis, which is characterized by variability of clinical manifestations, forms, and variants. Limited efficiency of antitumor immune protection in the patient plays an important role in progression of this disease. Survival of myeloma cells is promoted by some growth factors, including a number of interleukins. Cytokines and chemokines are secreted in response to intercellular interactions and stimulate tumor growth, inhibition of osteoblasts and increase of the osteoclastic activity. Cytokine genes show a significant allelic polymorphism. A single gene may exhibit numerous polymorphic sites located in exons, introns and promoter regulatory areas. Single nucleotide substitutions in the promoter region of cytokine genes are known to have a huge impact upon secretion and biological activity of these factors. Therefore, a study of allelic gene variants determining the levels of cytokine production will allow of establishing new immunogenetic factors associated with a high risk of disease development, including multiple myeloma. We have studied single nucleotide polymorphism in cytokine genes (IL-1α -889 TT, IL-1β +3962 TT, IL-6 -174 GG, and IL-6 nt565 GG), and clinical laboratory parameters (serum levels of albumin, β2-microglobulin, and hemoglobin) determining severity grade of multiple myeloma in 80 patients living in the North-Western region of Russia. It was found that the presence of certain cytokine gene variants, i.e., IL-1α -889 TT, IL-1β +3962 TT, IL-6 -174 GG, IL-6 nt565 GG or IL-1α -889 TT, IL- 1β +3962 TT or IL-6 -174 GG, IL-6 nt565 GG was associated with low albumin levels (< 3.5 g/DL), and high levels of β2-microglobulin (> 5.5 mg/l). A combination of all the four negative variants in homozygous state (IL- 1α TT -889, IL-1β +3962 TT, IL-6 -174 GG and IL-6 nt565 GG) increases the chance of six-fold reduction of albumin levels (p < 0.05); combinations of homozygous IL-1α TT -889, IL-1β +3962 TT, IL-6-174 GG. and IL-6 nt565 GG are associated with increased chance of high-level β2-microglobulin (> 5.5 mg/l) by more than two times. This data allow to consider IL-1α -889 TT, IL-1β +3962 TT, IL-6 -174 GG, and IL-6 nt565 GG genotypes additional negative immunogenetic factors in the prognosis of multiple myeloma.

Despite numerous attempts to control the course of chronic rhinosinusitis with nasal polyps (CRSwNP) by means of pharmacological treatment and new surgical approaches, the majority of patients experience lifelong persistence of this disorder, at recurrence rates of 50-60% within 18 months after surgical treatment. Since CRSwNP is a chronic persistent inflammatory process, it affects the entire body condition, including the state of systemic immune response. An elevation of NK (CD3-CD16+CD56+), activated NK (CD8+CD3-), NKT cells (CD16+CD56+CD3+), Treg (CD4+CD25brightCD127low to neg) cells and activated T-lymphocytes (CD3+CD25+) was revealed elsewhere among all the patients with CRSwNP, using a flow ytometry method. There was no difference between various disease phenotypes. We analyzed the status of cellular component of systemic immunity, dependent on clinical course of the disease and efficiency of the administered therapy of CRSwNP. The patients were divided into three subgroups. The follow-up period was 1 year. The first group comprised the patients who showed positive dynamics after conservative therapy, resulting into regression of nasal polyps and their grade than a year ago. The second group included the patients in whom the size of polyps remained the same. The third group included the patients with higher incidence of nasal polyps than a year ago.
We have shown a decrease of Treg, NKT cells, NK and activated NK, cytotoxic T-lymphocytes (CD3+CD8+), activated T-cell numbers in clinical group 3 with aggressive growth of polyps and low effect of standard therapy, which may cause deterioration of the immune system cellular populations, accompanied by presence of persistent productive inflammatory process of nasal cavity and paranasal sinuses. In the second group, a significant elevation of total lymphocyte number, total and activated T cells, T helpers (CD3+CD4+), cytotoxic T lymphocytes, NK and NKT cells was shown. Meanwhile, a decrease in absolute number of activated NK was observed despite the NK growth. Therefore, we can assume that the mechanism of their activation was disturbed and compensated by production of NKT cells and cytotoxic T lymphocytes. Moreover, we have shown in this group that the absolute number of Treg cells is increased; and these cells had a suppressive influence on effector cells of adaptive immune response, thus inducing incomplete elimination of infectious agents, which contribute to permanent incomplete course of inflammatory process. Chronic inflammatory process in CRSwNP affects systemic cellular immunity depending on the morbidity characteristics in the course of pathological process. The maximal intensity of systemic cellular immunity is observed in the group of patients that require permanent basic drug therapy. In case of aggressive CRSwNP and failure of standard drug therapy, we observed a decrease in absolute numbers of effector cells, along with decreased Treg lymphocyte numbers which may explain inefficient immune regulation of inflammatory process and medical interventions in this group of patients.

Juvenile idiopathic arthritis is a chronic inflammatory disease of the joints in children, mainly of autoimmune or auto-inflammatory nature. It is a heterogeneous group, which includes different subtypes of the disease. Different mechanisms may play role in the pathogenesis of distinct subtypes of juvenile arthritis. However, a long-term imbalance of pro- and anti-inflammatory cytokines is important for all subtypes of disease. The aim of the present study was to determine spontaneous and stimulated anti-inflammatory cytokines production by peripheral blood cells from the children with juvenile idiopathic arthritis. Patients of 2 to 17 years old with different subtypes of juvenile idiopathic arthritis (n = 99) and healthy children without signs of autoimmune diseases (control, n = 31) were examined. Spontaneous and phytohemagglutinin-stimulated concentrations of IL-1ra, IL-4, IL-10, TGF-β in supernatants of whole-blood cultures were determined by ELISA. Differences in the spontaneous and mitogen-stimulated secretion of the cytokines in patients with different subtypes of juvenile arthritis have not been revealed. The spontaneous IL-1ra, IL-4 and IL-10 production by blood cells in the common group of patients with juvenile idiopathic arthritis was similar to the controls. The median value of spontaneous TGF-β concentration in the patients was below the detection level, whereas blood cells of healthy children had a higher potential of spontaneous TGF-β production. IL-4 and IL-10 production after incubation of peripheral blood cells with phytohemagglutinin in patients and in the control group did not differ from the controls, while IL-1ra and TGF-β synthesis was significantly lower than in healthy children.
The spontaneous and/or stimulated production of IL-1ra, TGF-β by blood cells in children with juvenile idiopathic arthritis reflects the pathogenic significance of these cytokines in disease. Stimulation of cells can reveal a latent deficiency in the synthesis of cytokines, which is not evident when determining its concentration in serum or supernatants of spontaneous whole-blood cultures.

Investigation of the cytokine expression dynamics as well as the cytokine-producing potential of immune-competent cells allows extensive studies of their functional characteristics. mRNAs encoding a number of cytokine genes are relatively stable, thus their level may be used as a marker for assessing the levels of activation and proliferation of immunocompetent cells as well as for evaluating the cytokine-producing potential of immunocompetent cells.

In our work, we assessed correlations between the levels of mRNA expression specific for IL-10, TNF-α, GM-CSF cytokines determined in a culture of differentiated macrophage U937 cells, and protein concentrations of the same cytokines as measured in supernates of U937 cell cultures, without and after exposure to polyclonal activators. The IL-10, TNF-α, GM-CSF mRNA expression was determined by real-time quantitative polymerase chain reaction. Protein concentrations of IL-10, TNF-α, GM-CSF cytokines in the culture supernatant of U937 cells were measured by an enzyme immunoassay. The use of an initially homogeneous cell culture in the study is convenient due to the identical conditions in all experimental variants.

The most pronounced effect of polyclonal activators is exerted upon production of GM-CSF mRNA, as well as protein concentration of this cytokine in the cell culture supernatants, thus actually coinciding with RT-qPCR results.  The TNF-α mRNA level decreased under the influence of polyclonal activators, whereas concentration of this cytokine was decreased in the cell supernate. The TNF-α protein in a culture medium did not reflect temporal changes in the cellular TNF-α mRNA expression, probably, due to potential decrease of cellular mRNA occurring by the feedback inhibitory mechanism. While the cytokines can accumulate and remain in the supernatant, the mRNA-related events leading to cytokine formation may be completed earlier. Therefore, the signalling pathways and cytokine release kinetics should be studied after establishing the time dependence at short time intervals, which may be individual for each cytokine.

Thus, the results of a study using polyclonal activators suggest that polyclonal activators applied as mitogens, have a significant effect upon the concentration of secreted IL-10, TNFα and GM-CSF. In this case, polyclonal activators affect the levels of mRNA encoded by cytokine genes, thus indicating transcriptional mechanisms of its action. But, in view of the fact that the data are ambiguous, in order to achieve greater correspondence between the changes in the studied proteins and specific mRNAs, a detailed description of the time dependence is required for the changes in mRNA contents.

An inflammatory process accompanied by the changes in mucosal microbiota is underlying the gastroduodenal diseases. Chronic inflammation of gastric mucosa is developed and supported by secretion of pro-inflammatory and anti-inflammatory cytokines by epithelial cells and immune cells induced by Helicobacter pylori and other microorganisms. Under the conditions of dysbiosis and immune dysregulation, gastric epithelial layer becomes like intestinal or colonic epithelium. The aim of this work was to determine diagnostic and prognostic value of cytokines in intestinal metaplasia of gastric epithelium. 
204 patients with exacerbation of chronic gastritis, gastric ulcer, gastric polyposis and 40 healthy volunteers were included into the study, under their informed consent. The gastric biopsies were sampled by means of esophagogastroduodenoscopy (for histological and microbiological examination), along with drawing 5 ml of venous blood with serum separation (for enzyme immunoassay). Serum cytokine levels were studied by solidphase enzyme immunoassay. In statistical evaluation of the results, we have calculated sensitivity, specificity of indexes, logistic regression equations, characteristic curves were built with the definition of the index of consistency of the model by the area under the curves (AUC).
Histological examination of gastric biopsies showed features of intestinal metaplasia in 61 patients (29.90%), colonic metaplasia was found in 40 cases (19.61%), being absent in healthy volunteers. The greatest sensitivity of intestinal metaplasia was observed for plasma levels of interleukin (IL)-6, IL-4, erythropoietin (EPO), tumor necrosis factor (TNF)α, IL-18, vascular endothelial growth factor (VEGF), interferon (IFN)α levels; in colonic metaplasia, for receptor antagonist IL-1β (IL-1ra), IL-8, EPO, IL-18, monocyte chemoattractant protein (MCP)-1, VEGF, IFNα, IL-1β, IL-6, IL-17. An increase in IL-6, EPO, IL-18, VEGF, IFNα were also common, thus indicative for changed functional activity of cytokines due to microbial contamination of gastric mucosa, tissue hypoxia with activation of angiogenesis, confirming a transition of intestinal metaplasia to gastric carcinogenesis. The greatest specificity in intestinal metaplasia was observed for IL-1β, IL-1ra, IL-8, IL-17, IL-2, IL-10; in colonic metaplasia, for IL-18, IFNα, IL-4, МСР-1, VEGF. In the intestinal metaplasia, the AUC interval was higher than 0.7 for IL-2, higher than 0.65, in VEG; in colonic metaplasia > 0.91, for IL- 18, VEGF, МСР-1, IFNα, having a significance level of < 0.001. The obtained prognostic models of intestinal metaplasia of gastric epithelium, according to the AUC index, had very good (Table 2, formula 1) and excellent quality (Table 2, 3, formula 2-11), confirmed by a high percent of cases which were correctly classified of metaplasia.
   Determination of serum cytokines in intestinal metaplasia of gastric epithelium is of diagnostic and prognostic value, and should be used for early diagnosis of precancerous conditions of the gastric mucosa, both as single indexes (IL-2, VEGF), and combined indicators, according to the calculated logistic regression equations.

It is known at the present time that immunological biomarkers may become more sensitive, non-invasive methods of graft rejection diagnosis than those currently used. A growing amount of studies in animal models shows that chemokines, as active participants in the immune process, may be used to this purpose. Our earlier studies have shown an important prognostic significance of IL-6, IL-2, 17A and IL-1RA increase in pre-operative period as markers of acute kidney allograft rejection. When assessing changes in studied peripheral blood growth factors, we concluded that a sharp decrease in BDNF content is a diagnostically significant early sign of kidney allograft rejection. The aim of this study was to identify the prognostic role of serum chemokine levels at the preoperative stage, taking into account the production of anti-HLA antibodies during the post-transplant period as a risk factor of kidney allograft rejection. A comparative analysis of chemokine serum concentrations was performed in the patients with terminal-stage chronic kidney disease (CKD). In the patients from main clinical groups, the blood cytokine levels were measured 6 hours before transplantation, i.e., Eotaxin (CCL11), GRO-α (CXCL1), IL-8 (CXCL8), IP-10 (CXCL10), MCP-1 (CCL2), MIP-1α (CCL3), MIP-1β (CCL4), SDF-1α (CXCL12), RANTES (CCL5), MIG (CXCL9) by means of multiplex immunological assays, using appropriate test systems. The studies have shown significant changes in several chemokines in the CKD patients compared to age-matched controls. However, the following diagnostically significant biomarkers associated with early rejection of transplanted kidney should be considered: increased concentration of CCL2 and CCL4 chemokines, as well as an acute decrease in CCL11. Significantly decreased CXCL12 concentration in peripheral blood could be considered a marker of favorable posttransplant clinical course.  Occurence of HLA antibodies in recipients is also associated with elevated serum levels of CXCL8, CXCL10, CCL4, and CCL5.

Ischemic stroke is the most common form of brain stroke. It is associated with functional changes of various blood and bone marrow cell populations, altered release of various cytokines, chemokines, etc. There are conflicting data about serum and plasma TNFα levels in acute ischemic stroke.
We have examined 21 patients with a diagnosis of ischemic stroke treated at the hospital. The severity of ischemic stroke was evaluated by neurologists, in accordance with NIHSS criteria at admission and at discharge. 
In the patients with ischemic stroke, we have found a significantly increased serum concentration of tumor necrosis factor-α (p < 0.001), as compared with healthy individuals. The highest concentrations of this marker were recorded on days 1 and 3 of the disease, being significant at p < 0.001 and p= 0.003, respectively, then decreasing by day 14, however, not reaching, the levels of control group. It should be noted that, among patients with ischemic stroke, there is a significantly (p < 0.001) increased proportion of individuals with high serum concentrations (>10 pg/ml) of this cytokine, i.e., 76.2±9.3% on day 1 of the disease.
To statistically evaluate the individual differences of the patients’ dynamics, they were divided into subgroups, depending on the level of TNFα on the 1st day of hospitalization, using a discriminant analysis with estimation of a classification matrix. The correlation analysis showed numerous strongly positive correlations between TNFα levels on the 1st and 3rd days, as well as between similar indexes on the 1st and 14th days. A correlation between TNFα concentrations on the 3rd and 14th day was also found (r = 0.711; p < 0.01). Also, positive correlation in various periods of observation was established between the absolute levels of cytokine and differences in their concentration changes. The level of tumor necrosis factor-alpha on the first and third day of hospitalization did positively correlate (respectively, r = 0.503, p < 0.01; r = 0.411, p < 0.01) with the volume of the ischemic lesion according neuroimaging methods research. The volume of the ischemic focus was positively correlated with the difference in the concentration of TNFα on days 1-3 and 1-14 (respectively, r = 0.425, p < 0.01; r = 0.507, p < 0.01).
The results of our study show a necessity for measuring TNFα levels at admission, in order to plan treatment in these groups of patients, especially in cases of increase or insufficient decrease in TNFα recorded on the 3rd day of therapy.

Uveal melanoma is a malignant neuroectodermal tumor. Despite of radical primary treatment with the use of enucleation of the eye or radiation therapy, almost in 50% of patients develop metastatic disease. The effectiveness of various methods of treatment of malignant tumors, the prognosis of the disease largely depend on the state of the patient's immune system. Data on the ratio of lymphocytes of different populations, their relationship with the stages of uveal melanoma are currently not available. 84 patients with uveal melanoma were examined (44 women and 40 men; mean age 53.7 ± 12.2 years). In group 1 (small tumors) included 36 patients, in the 2nd (medium-sized tumors) - 26 patients, 22 patients made up the 3rd group (large tumors). The control group included 33 practically healthy donors. The material of the study was samples of whole blood taken from the ulnar vein on an empty stomach in the morning with the help of vacuum systems in test tubes Vacuette® with anticoagulant K3EDTA. Immunophenotyping was performed using flow laser cytofluorimetry using the Multitest 6-Color TBNK Reagent monoclonal antibody system in BD TruСount test tubes (Becton Dickinson, USA), BD FACSCanto II cytometer (Becton Dickinson, USA). Erythrocyte lysis and leukocyte fixation were performed using a BD FACS ™ Lysing Solution lysing solution (Becton Dickinson, USA). The relative and absolute levels of lymphocyte populations and subpopulations were determined in the Canto program (Becton Dickinson, USA), with the selection of the analyzed region in the general population expressing CD45 + antigen and cell granularity (CD45 + PerCP-Cy5,5 * / SSC); fluorochromes labeled antibodies to CD3 + (FITC), CD4 + (PE-Cy7 *), CD8 + (APC-CY7 *), CD16 + / 56 + (PE), CD19 + (APC *) were used to differentiate: T-lymphocytes (CD3 +), T-helper cells (CD3 + CD4 + CD8-), T-cytotoxic (CD3 + CD4-CD8 +), T-double positive (CD3 + CD4 + CD8 +), NK-cells (CD16 + CD56 +), B-lymphocytes (CD19 +) , calculate the ratio of CD3 + CD4 + / CD3 + CD8 + subpopulations - the index reflecting the balance of T-helper cells and cytotoxic T-cells (CD4 + / CD8 +). Our individual analysis of the composition of lymphocytes allows us to conclude that the growth of uveal melanoma is accompanied by systemic multidirectional changes in the qualitative and quantitative composition of immunocompetent cells, affecting both innate (NK cells) and adaptive (T-lymphocytes) links of antitumor immune defense. The results are important for the development of personalized approaches to the prognosis and treatment of patients with uveal melanoma. The dynamics of changes in the quantitative composition of lymphocyte subpopulations in oncological diseases may be of value for monitoring immunotherapeutic effects and supplement the clinical assessment of the course of the underlying disease.

 

 

 

It is known that functional activity of complement system depends not only on balance and concentration of components participating in formation of the system end products, but also on levels of inhibitory activities. Numerous relations with hemostasis also substantially contribute to general level of complement system activity. Changes in complement system functioning are inevitable during chronic diseases accompanied with immune system dysregulation. All mental diseases tend to be chronic and are they aggravated by patients’ immune system changes. Autism spectrum disorders in children is a group of mental disorders. Immune system dysregulation is usually detected in such patients, manifesting as excessive susceptibility to viral and bacterial infections. Therefore, the level of its functional activity is diagnostically and prognostically significant in this pathology, since the complement system is a key element of immune system.
We have evaluated functional activity of complement system in patients with autistic spectrum disorders, using the method which was developed earlier. It is based on the reaction of the protozoa (Tetrahymena pyriformis) which are both targets and activators for the complement system. The complement system capacity (cSC) was used as the main parameter of complement evaluation. The half-time of protozoa survival (T50) was defined using the BioLat device for each serum specimen added at four concentrations (1/20, 1/40, 1/80, 1/160 dilution). The complement capacity was calculated as the area enclosed by influence curve of the reciprocals of T50 and the serum dilution. According to Mann–Whitney U test, the difference between patients’ and healthy volunteers’ groups was established as Z = 4.43 (by T50 at 1/160 dilution), p < 0.001 and by cSCas Z = 5.8, p < 0.001. cSC was calculated from the results obtained at each serum concentration measured. The difference between the two groups according to Mann–Whitney U test appeared to be more significant than the difference according to T50. Therefore, cSC was taken as the main characteristic of complement system function.
The contribution of hemostasis plasma components to complement system functional activity level was estimated by determination of complement capacity in plasma and serum of each blood sample from 6 patients with autism spectrum disorders and 5 healthy donors. All healthy donors showed small difference between plasma and serum complement capacity, and their complement activity was higher in plasma. In patients’ group, the complement capacity levels in plasma and serum differed significantly. The cSC levels of two patients were higher in serum than in plasma, and the cSC levels of three other patients were significantly higher in plasma than in serum. Differential involvement of coagulation into the complement system activation may be indicative for the immune system dysfunction which is observed in patients with autistic spectrum disorders of different etiology.

Autoantibodies recognizing the cancer-retina autoantigen called recoverin (RCVRN-AutoAb) may serve as a highly specific biomarker of cancer-associated retinopathy. However, they may also be found in some cancer patients without clinical evidence of retinopathy. In the present study, dot-ELISA and Western blot assays were used to demonstrate the presence of circulating RCVRN-AutoAb in 4/7 (57%) of patients with recently recognized pathological entity, non-invasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP); other thyroid tumors represented by follicular adenomas, and classical and follicular variants of papillary thyroid carcinomas, demonstrated low frequencies of RCVRN-AutoAb (0/15, 1/20 (5%) and 1/15 (7%), respectively), with no significant differences from healthy individuals (0/15). Our data implicate the circulating RCVRN-AutoAb as a potential biomarker of NIFTP capable of discrimination of this novel pathological entity from other thyroid tumors.

Bronchial asthma (BA) is the most widespread chronic inflammatory disease. Since BA is associated with a systemic inflammation state, a comprehensive study of its effect in this disease, and influence of pathogenetic therapy should be performed, by studying the whole blood cytokine status of the patients suffering with BA. The cells from respiratory tract in acute-phase BA patients may produce pro-, as well as anti-inflammatory mediators. The anti-inflammatory mediators are able to suppress activity of immune cells in peripheral blood. Thus, the aim of present study was to evaluate eventual inflammation-associated and functional activity of immune cells from the patients’ peripheral blood in BA and following appropriate therapy. Bacterial lipopolysaccharide (LPS) a classical pro-inflammatory agent. We have studied an LPSinduced cytokine-induced ex vivo secretion model by peripheral blood immune cells, as a relevant test for their functional activity. The LPS-induced responses of whole blood cells from patients with proven BA diagnosis have been studied at pre-treatment time points, and following two weeks of basic anti-inflammatory therapy. According to clinical indications, the antagonists of CysLTR1, or combinations of glucocorticosteroids and β-adrenoreceptor agonists were administered by inhalation to BA patients. LPS-induced production of TNFα, IL-6, IL-8 (at 6 h) and IFNγ, IL-17A or IL-1β (at 24 h) by whole blood cells from BA patients or healthy volunteers has been assessed by ELISA technique. The cytokine production from non-stimulated whole blood cells from BA patients and healthy volunteers were used as the baseline control. IL-4 concentrations in plasma of BA patients and healthy volunteers were also measured. We have shown a decrease of IL-6 production in control blood samples from BA patients after two weeks of therapy. This may indicate the attenuation of the observed inflammatory process. The therapy applied did not influence the background levels and LPS-induced secretion of IL-1β, IL-1ra, IFNγ, and IL-8 in whole blood samples from BA patients. IL-4 plasma levels in BA patients were not changed after two weeks of therapy. It has been shown that whole blood from BA patients produced less TNFα and IL-8, both in control samples, and during their response to LPS, than the values obtained in healthy volunteers. These findings are in agreement with a notion that BA causes partial depression of innate immune cells activity. The increased LPS-induced TNFα secretion by the whole blood cells from BA patients has been observed following two weeks of basic anti-inflammatory therapy. We suggest that the increased LPS-induced TNFα secretion could be explained by partial restoration of peripheral blood immune cell activity associated with anti-inflammatory BA therapy. To elucidate the mechanism of increased LPS-induced TNFα secretion, we have estimated whole blood concentration of soluble CD14 (sCD14) in BA patients. No significant differences between sCD14 concentrations have been found. Obtained result presume existence of sCD14-independent mechanism of TNFα regulation by whole blood cells in response on LPS which may occur during anti-inflammatory therapy of BA. We suppose that basic anti-inflammatory therapy of BA does not simply reduce IL-6 concentration in peripheral blood, but may also partially restore the activity of innate immune cells in BA patients.