Tumor microenvironment (TME) is formed as a result of interaction and cross-linking between the tumor cell and different types of surrounding cells. Recent studies have shown that the tumor reprograms the microenvironment so that TME promotes the development of primary tumors, their metastasis and becomes an important regulator of oncogenesis. Under the influence of the tumor, the immune profile in the TME undergoes significant changes, “editing". An immunosuppressive network is formed, which suppresses the activity of the main effector of cellular immunity — T lymphocytes. T cells in TMA are in a state of anergy and exhaustion. T cells in TME are characterized by increased expression of inhibitory receptors, decreased secretion of cytokines and cytolytic activity. Blocking inhibitory receptors with specific antibodies can lead to the restoration of the functions of exausted T cells. Therefore, the restoration of the functional activity of T lymphocytes is one of the important strategies in cancer immunotherapy. The formation of the immune profile is influenced by genetic aberrations accumulating in the tumor. They play an important role in creating a specific, characteristic only for this tumor immune environment in the TME. Genetic changes in tumor cells lead to phenotypic and functional rearrangements of lymphocytes, which allows the tumor to escape the reaction of immune cells. Since many tumors occur after prolonged inflammation or exhibit characteristics of chronic inflammation as they progress, inflammation is considered an important factor in the formation of immune profile in TME. Immune infiltrates from different human tumors associated with inflammation may contain valuable prognostic and pathophysiological information. Macrophages in the TME now began to be regarded as descriptive marker and as a therapeutic target. One of the main mechanisms by which tumor cells reprogram surrounding cells is the release of exosomes — small vesicles that carry and deliver proteins and nucleic acids to other cells. When exosomal cargo is absorbed, molecular, transcriptional and translational changes occur in the recipient non-tumor cells in the TME. Therefore, tumor exosomes are an effective means by which the functions of immune cells in TME are purposefully changed. Thus, along with individual molecular and genomic testing of the tumor, attention should be paid to a deeper analysis of the immune profile of TME. It is a large resource of biomarkers and targets for immunotherapy.

Mortality from diseases of the circulatory system is a challenge for the modern health care. Arterial hypertension (AH) mostly contributes to development of cardiovascular complications. It often proceeds against the background of metabolic disorders. Pathogenesis of hypertension is currently being considered a multifactorial disease. Pathogenesis of hypertension certainly has distinct features in presence of metabolic disorders,. Therefore, it is relevant to summarize current literature on the role of immunological disorders, endothelial dysfunction and hemostatic disorders in AH genesis during metabolic syndrome (MS). Most authors agree with existence of several mechanisms that determine relationships between AH and insulin resistance. Development of hypertension in MS patients with is a consequence of immunometabolic processes. Abdominal obesity is an important component of MS. It is associated with chronic inflammation of visceral adipose tissue, its excessive infiltration by immune cells, and increased production of adipokines and cytokines (TNFα, IL-6) with hypertension. AH is associated with a significant increase in T cells, that mediate endothelial dysfunction (ED) and provide a link between hypertension and subsequent atherosclerosis. T lymphocytes trigger a cascade of reactions. IL-17 is the end product of these events It is involved not only in increasing blood pressure, but also contributes to the development of vascular wall stiffness in АН patients. Thus, the relationship between several types of immune cells leads to inflammatory reactions, including those of vascular wall, initiating endothelial dysfunction. Chronic non-specific inflammation in MS, supported by the cytokine system, is a triggering mechanism for ED progression. Excessive production of endothelin-1 and inhibition of nitric oxide production are the classic markers of ED. Immune damage leads to imbalance in the production of vasoconstrictor and vasodilating substances, proliferative and antiproliferative factors in endothelium. It was shown that ED is an integral aspect of the insulin resistance syndrome in pathogenesis of arterial hypertension associated with metabolic disorders, and contributes to its worsening, increased vascular reactivity and further AH development. According to modern studies, it has been shown that excessive synthesis of pro-inflammatory cytokines introduces disturbances in the system of vascular hemostasis. When studying the effects of metabolic disorders upon hemostatic system, we may conclude that activation of fibrinolytic and plasma chains occurs in the same way for both men and women, with small gender characteristics of individual components. The rheological properties of the blood are also changed with developing MS. Systematization of the available literature data on the issue under study can serve as a basis for determining prognostic criteria of hypertension progression and risk of thrombotic complications.

Development of angiogenesis depends on the functional state of endothelial cells, as well as on the balanced secretion of cytokines, growth factors and chemokines by endothelial cells and cells of microenvironment. Macrophages represent an essential component of the microenvironment and take part in the formation of blood vessels both due to the production of cytokines and due to contact interactions with endothelial cells. VEGF is among the most important cytokines that control angiogenesis at all its stages. Currently, the role of VEGF in the intercellular interactions of endothelial cells and macrophages is not well described. The aim of our study was to investigate the effect of VEGF deprivation using monoclonal antibodies on angiogenesis under conditions of co-cultivation of endothelium and macrophages. Materials and methods: monoclonal antibodies to VEGF-A were used for VEGF deprivation in monoculture of endothelial cells and in co-culture of endothelial cells with macrophages. The IL-1β, IL-6 and TNFα cytokines were used as inducers. When VEGF-A was removed from the medium, endothelial cells show plasticity and form longer vessels, they modify the expression of VEGF receptors. Macrophages regulate endothelial cell activity through the secretion of cytokines, including VEGF, and through contact interactions with endothelial cells. THP-1 cells increase the sensitivity of endothelial cells to VEGF by stimulating the VEGFR1 and VEGFR3 expression, this effect is VEGF-A-independent. The IL-1β, IL-6, TNFa cytokines independently stimulate non-branching angiogenesis, increasing the length of the vessels. At the same time, IL-ip increases the VEGFR1 expression on the surface of endothelial cells. In contrast, IL-6 and TNFα decrease it, thereby regulating the sensitivity of endothelial cells to VEGF. The effects of these cytokines are not dependent on VEGF-A. The IL-1β, IL-6, TNFα cytokines promote acquisition of anti-angiogenic properties by THP-1 cells that is independent on VEGF-A, as well as on expression of its receptors by endothelial cells. Thus, VEGF is an important, but not the sole factor controlling angiogenesis. Under conditions of VEGF-A deficiency, either endothelial cells or microenvironment cells are able to compensate for its functional load due to the production of other growth factors.

Microvesicles (MVs) are small (100-1000 nm) subcellular structures produced by both motionless and activated cells that can transfer molecules to target cells, and regulate physiological and pathological processes. MVs of leukocyte origin, in particular those produced by natural killer cells (NK cells), remain the least studied population of MVs. NK cells can change the functional activity of endothelial cells (ECs) and are involved in regulating angiogenesis. The ability of NK cell-derived MVs to influence the functionality of ECs is understudied currently. We aimed to study the effect of MVs produced by NK cells of the NK-92 cell line on the phenotype, caspase activity, proliferation and migration of ECs of the EA.Hy926 cell line. We cultured ECs in the presence of MVs derived from the NK-92 cell line, and then used flow cytometry to assess changes in EC phenotype, intracellular protein transfer from MVs to ECs, and the relative death of ECs. We used western blot analysis to evaluate the expression of granzyme B in NK cells and in the MVs that they produced, as well as the expression of granzyme B, caspases, extracellular-regulated kinase (ERK) and protein kinase B (AKT) in ECs. We also assessed the proliferation and migration of ECs in the presence of MVs derived from cells of the NK-92 cell line. The results revealed significant differences in the proteomic profiles of cells of the NK-92 cell line and their MV product. Contact between ECs and MVs derived from cells of the NK-92 cell line is accompanied by the following events: a) expression of granzyme B in ECs, b) activation of caspase-9 and caspase-3, with partial EC death, c) appearance of the panleukocyte marker CD45 on ECs, d) decrease in CD105 expression, and increase in CD34 and CD54 expression, and e) inhibition of EC migration. Transfer of ERK (but not AKT) from MVs derived from cells of the NK-92 cell line to ECs, at a concentration 10 times lower than that which causes EC death, leads to an increase in EC proliferation.

Peripheral blood monocytes are heterogeneous CD14⁺ cell population, some of which also express CD16 molecule. Differences in phenotype between monocyte subpopulations can affect their functional activity, as well as the ability to further differentiate into dendritic cells (DCs). DCs are professional antigen-presenting cells which induce the immune response or, conversely, maintain the immunological tolerance. The aim of the present study was to analyze the relationship between monocyte subpopulations and the functional activity of monocyte-derived DCs, as well as DC sensitivity to the tolerogenic effect of dexamethasone. DCs were generated by cultivating enriched fractions of CD14⁺ monocytes with or without CD16⁺ cell depletion (CD16^-Mo-DCs or CD16⁺Mo-DCs, respectively) in the presence of IFNα and GM-CSF. Monocyte subpopulations were obtained by immunomagnetic negative selection. CD16⁺Mo-DCs were characterized by lower ability to take up FITC-dextran and higher allostimulatory activity compared to CD16^-Mo-DCs. In addition, CD16⁺Mo-DCs showed higher apoptosis-inducing activity against autologous CD4⁺T lymphocytes and allogeneic CD8⁺T lymphocytes, but were similar to CD16-Mo-DCs in their ability to induce apoptosis in allogeneic CD4⁺T lymphocytes. TNFa production level, similar for both types of DCs, was negatively correlated with CD16^-Mo-DC allostimulatory activity and directly correlated with apoptosis-inducing activity of CD16⁺Mo-DCs towards allogeneic CD4⁺T cells. CD16-Mo-DCs and CD16⁺Mo-DCs were similar by their IL-10 production, which was inversely related to allostimulatory activity of both types of DCs. Dexamethasone increased endocytic activity, decreased the ability to stimulate autologous and allogeneic T cells, inhibited TNFα production of CD16^-Mo-DCs and CD16⁺Mo-DCs. However, CD16⁺Mo-DCs demonstrated a more pronounced increase in endocytic activity and more dramatic decrease in their ability to stimulate the proliferation of CD4⁺T cells in auto-MLR. Also, addition of dexamethasone into CD16⁺Mo-DCs cultures led to the increase in DC pro-apoptogenic activity against autologous CD8⁺T lymphocytes. Thus, the presence of CD16⁺ cells among monocyte population affects the properties of IFNα-induced monocyte-derived DCs and DC sensitivity to the immunomodulatory effects of dexamethasone.

The discordant immunologic response to antiretroviral therapy in HIV-infected patients is characterized by ineffective recovery of CD4⁺T cell counts. The role of regulatory T cells in discordant response to the treatment remains poorly understood both due to the lack of specific and reliable markers of regulatory T cells and their subset’s heterogeneity. In the present work, we studied two groups of HIV-infected patients receiving antiretroviral therapy for more than two years and thus having their viral load suppressed (less than 50 copies of HIV per ml of blood): those who responded (n = 22) and did not respond (n = 19) to the treatment with an increase in their CD4⁺T cell counts. The control group consisted of uninfected volunteers (n = 23). The CD4⁺T lymphocyte subset composition was examined by flow cytometry. It was shown that in HIV-infected patients with ineffective immune recovery compared with subjects having a standard response to antiretroviral therapy, the absolute counts of regulatory T cells, as well as CD4⁺T lymphocytes, was reduced in all maturational subsets: naive cells, central memory, effector memory, and terminally differentiated effectors. That differed immunological nonresponders from patients with a standard response to the treatment, which had a shortage only in naive and central memory regulatory T cell subsets. It is important to note that in HIV-infected patients with a discordant response to therapy, the proportion of effector memory regulatory T cells, that posses the most prominent suppressive capacity, was significantly increased compared with that in other CD4⁺T lymphocyte subsets. Apparently, despite of regulatory T cell deficiency, in HIV-infected patients with a discordant response to the treatment, the regulatory T cell pool size is big enough to control CD4⁺T lymphocyte activation. Nevertheless, the number of regulatory T cells may not be sufficient to suppress the over-activation of immunocompetent cells that are not in the CD4⁺T lymphocyte subset. This can partly explain the increased cell activation level in patients with a discordant response to therapy as compared with those who have a standard respond to the treatment.

Hepatitis B is an infectious viral disease in which damage or destruction of liver tissue occurs, and which can turn into a chronic form. In chronic hepatitis B (CHB), hepatocytes are replaced by connective tissue as they die, resulting in fibrosis, and then cirrhosis of the liver. Early diagnosis of liver fibrosis is an urgent task in the development of CHB. Already at this stage of the disease, the immune system is activated, which plays a leading role in liver damage in CHB. The main regulators of immune processes are cytokines, which mediate intercellular interactions. A separate group of cytokines are chemokines — proteins of cell migration. In CHB, they are responsible for infiltration of liver tissue by activated white blood cells. Cytokines and chemokines are active participants in fibrogenesis, so they can serve as biomarkers for the development of liver fibrosis, including in the early stages. The purpose of this study was to analyze the content of certain cytokines/chemokines in the peripheral blood of patients with CHB in order to search for potential biomarkers of the initial stages of liver fibrosis. The study included 30 patients with a confirmed diagnosis of CHB with stages of liver fibrosis F0-F1 on the Metavir scale, 36 patients with a diagnosis of chronic hepatitis C (F0-F1) and 37 conditionally healthy individuals as a control group. Concentrations of the following cytokines/chemokines were determined: IFNγ, TNFα, CCL2/MCP-1, CCL8/MCP-2, CCL20/MIP-3α, CXCL9/MIG, CXCL10/IP-10, CXCL11/I-TAC by multiplex analysis using xMAP technology (Luminex, USA). As a result of the study, it was found that in patients with CHB, the plasma content of cytokine TNFα and chemokines CCL2/MCP-1, CXCL9/MIG and CXCL10/IP-10 was increased, and the chemokine CCL8/MCP-2 was reduced, which indicates the possibility of using these cytokines/chemokines as biomarkers of liver damage in CHB. In the examined group of patients with CHB, there was no dependence of the concentrations of cytokines and chemokines in the blood plasma on the viral load, which may be explained by its low level. For plasma cytokines in patients with CHB, correlations were found between TNFα and CCL2/MCP-1 and CCL8/MCP-2 chemokines , which was not observed in the control group. At the same time, in the control group, a correlation was found between the content of TNFα and the chemokine CXCL9/MIG, which was not detected in the group of patients. For both groups, a correlation was found between the content of CXCL9/MIG and CXCL10/IP-10 chemokines. Based on the study data, an algorithm has been developed that allows us to establish chronic hepatitis B or chronic hepatitis C as the cause of the initial form of fibrosis F0-F1 in terms of the content of cytokines IFNγ, CCL2/MCP-1 and CCL8/MCP-2 in blood plasma with a diagnostic efficiency of 89.4%.

Diabetic retinopathy (DR) is multifactorial by its origin, involving many cytokines and growth factors. Studies of cytokine levels in biological fluids seem to be relevant for an in-depth understanding of the disease pathogenesis. The purpose of the work was a comparative analysis of 45 cytokines at systemic (blood serum (BS)) and local (vitreous humor (VH)) levels in the patients with complicated proliferative DR, showing various features of the clinical pattern. The content of cytokines was tested in 53 samples of BS and 32 samples of VH in 53 patients with type 1 and type 2 diabetes mellitus with severe proliferative DR. We used the multiplex analysis technique by means of xMAP platform and Luminex xPONENT 3.1 program using 45-plex sets (Procarta Plex «eBioscience», Austria). 25 cytokines were detected at significant amounts in BS test samples, and 27 cytokines were revealed in VH specimens. Sensitivity limits of the test system allowed to find significantly higher levels of 7 cytokines (IL-6, IL-8, IP-10, MCP-1, HGF, LIF and VEGF-A) in VH samples, than in the BS, thus indicating to their local intraocular production. Correlations between the contents of VEGF-A growth factor and amounts of cytokines, including those involved in inflammatory reactions, are shown in VH, thus presuming the interrelation of pathogenetic components, i.e., inflammation and neoangiogenesis. The features of intraocular cytokine content were determined for various manifestations of diabetic ocular changes. Hemophthalmus has been shown to be associated with increased IL-8 and IP-10; iris rubeosis, with increase in LIF; proliferative DR activity was associated with higher MCP-1 levels, and extremely severe changes were related to increase in IL-6 and EGF. Testing of cytokines in biological fluids is informative when studying the mechanisms of inflammation, neoangiogenesis, and protective responses in pathogenesis of diabetic retinopathy.

Physical and chemical factors, obesity, lipid metabolism disorder, diabetes affect the state of the vascular endothelium, the processes of thrombus formation, fibrinolysis and increase the risk of developing cardiovascular diseases. It can be assumed that the development and course of pathological processes in the cardiovascular system, caused by vibration disease with the onset of diabetes, will undergo changes. The purpose of the study was to assess the state of heart condition according to the level of specific autoantibodies with the combined effect of vibration disease and type 2 diabetes mellitus. Patients with vibration disease (group I), patients with type 2 diabetes (group II) and persons with vibration disease in combination with type 2 diabetes (group III) were examined. Individuals do not have a history of coronary heart disease, stroke, and myocardial infarction have been included in obsledrovanie. Serum levels of specific autoantibodies characterizing the state of the heart have been studied. It was revealed that the content of autoantibodies to 1-adrenoreceptors in patients of group I was higher than in individuals of group II. The relative content of autoantibodies to the components of the membrane and cytoplasm of myocardial cells in patients of groups I-III did not differ. It was found that elevated levels of autoantibodies to components of the membrane and cytoplasm of myocardial cells, cardiomyosin, 1-adrenoreceptors were observed more often in patients with vibration disease combined with diabetes and in persons with vibration disease than in people with diabetes. Persons with a reduced content of autoantibodies to 1-adrenergic receptors were not detected among patients of groups I and III. Changes in the levels of specific autoantibodies in persons with vibration disease may indicate the development of functional metabolic and structural changes in the heart, disorders of its electrical activity that have not yet been manifested in the form of a pathological process. Lower levels of 1-adrenoceptor autoantibodies in patients with type 2 diabetes without vibration disease can be caused by increased levels of catecholamines, which is characteristic of diabetics. Further research, including clinical data and indicators of functional diagnostics is necessary to confirm our assumptions.

Great scientific interest to problems of obliterating atherosclerosis of vessels of the lower extremities is caused by increase in incidence of this pathology and multidisciplinary approach in diagnostics and treatment the research objective: to study interrelations between quantity of the circulating microparticles of lymphocytic origin and development of critical ischemia of the lower extremities. The executed research belonged to cohort prospective researches. Two studied groups were created: 1^st group included 75 patients with ischemia IIB degrees on Pokrovsky—Fontaine's classification. The 2nd group included 75 patients with clinical manifestations of critical ischemia. Also, the group of control of 75 almost healthy faces which do not have symptoms of obliterating diseases of vessels of the lower extremities was created. For objectification of assessment of degree of ischemia measured anklebone’s arterial blood pressure, the anklebone-humeral index, toe’s arterial blood pressure, Transcutaneous tension of oxygen in foot fingers. For blood-groove assessment in arteries of the lower extremities used ultrasonic doppler sonography of arteries of the lower extremities and the KT-angiography. The research was conducted at arrival of the patient in a hospital prior to therapy. Lymphocytes and the circulating microparticles defined according to the standard protocol. At almost healthy faces the number of the circulating microparticles of lymphocytic origin made of control group 385 (260479) on 100 lymphocytes. In 1^st group this indicator was twice higher and made 728 (654-836) (p₁ = 0.002). In the 2^nd group it was 5 times higher, than in group of control and authentically differed from an indicator of 1^st group (1570 (1124-2120) (p₁< 0.001, p₂ = 0.002) the Correlation coefficient between number of the circulating microparticles and anklebone’s arterial blood pressure r_S = -0.82 (p = 0.003), Correlation between the anklebone-humeral index and number of microparticles was equal to r_S = -0.92 (p < 0.001). Toe’s arterial blood pressure had average force negative correlation with number of microparticles. The transcutaneous tension of oxygen and number of microparticles had a correlation coefficient equal rS = -0.89 (p = 0.002). The correlation coefficient between a gleam of the general femoral artery and number of microparticles of lymphocytic origin made r_S = -0.44 (p = 0.016). A correlation coefficient between a gleam of a superficial femoral artery and number of microparticles of r_S = -0.64 (p = 0.002). Negative correlations of number of microparticles with the kept gleam of a popliteal artery — r_S = -0.79 (p < 0.001), and a back tibial artery — r_S = -0.86 were high, (p < 0.001). Very high negative correlation coefficient was between the kept gleam of a lobby tibial and number of microparticles of lymphocytic origin of r_S = -0.91 (p = 0.003). The revealed high and very high correlation bonds between number of the circulating microparticles of lymphocytic origin and indicators of a condition of an arterial blood-groove of the lower extremities, and the kept gleam of arteries of the lower extremities allow to speak about the prospects of use of microparticles as pathogenetic marker of critical ischemia of the lower extremities.

Forty women with gonarthrosis were included in this study. The main group consisted of 19 patients having osteoarthritis (OA) with metabolic syndrome (MS), the control group consisted of 21 patients with OA but without MS. It was found that metabolic phenotype of gonarthrosis, i.e. OA with concomitant MS, was different from OA without MS in terms of pain measured with visual analogue scale (VAS) (65 mm in the main group vs 47 mm in control group, р = 0.001) and other OA symptoms in accordance with Knee Osteoarthritis Outcome Scale (KOOS) (43.2 points in the main group vs 76.1 points in the control group, р = 0.001). These main distinguishing features were associated with low quality of life measured with non-specific questionnaire Short Form -36 (SF-36) (30 points in the main group and 40 points in the control) and clinically significant signs of depression, detected with Patient Health Questionnaire-9 (PHQ-9) (12 points in the main group and 7 points in the control group). The metabolic phenotype of gonarthrosis was characterized with laboratory features of low-grade systemic inflammation as evidenced by increased CRP (11.4 mg/ml in the main group vs 3.2 mg/ml in the control group, р = 0.03), IL-6 (2.6 pg/ml in the main group vs 0.7 pg/ml in the control group, р = 0.001), IL-18 (196.6 pg/ml in the main group vs 61.4 pg/ml in the control group, р = 0.001) in the peripheral blood serum, as well as increase in antibodies against Col2 (27.1 ng/ml in the main vs 5.5 ng/ml in the control group, р = 0.01) , and dyslipidaemia — increase in LDL-cholesterol (5.5 mmol/l in the main group vs 5.9 mmol/l in the control group, р = 0.032) and triglycerides (2.026 mmol/l in the main group and 1.36 mmol/l in the control gropu, р = 0.02). In conclusion, MS-associated OA phenotype occurs due to pathogenetic similarities between OA and MS (syntropy) based on systemic low grade inflammation. This OA phenotype is not well studied and needs further research to develop new treatments targeting these two comorbid disorders as a single disease.

The following specificcharacteristics of the composition of intestinal microbiota in patients with irritable bowel syndrome (IBS) were identified using a metagenomic analysis (16 S rRNA): 1) an increase in the representation of Actinobacteria, including Bifidobacterium spp., Firmicutes, including representatives of Streptococcaceae (Streptococcus), Lachnosperaceae (Dorea), Veillonellaceae (Dialister), Proteobacteria (Enterobacteriaceae and Desulfovibrionaceae families); 2) a decrease in the population of Bacteroidetes, including representatives of the families Prevotellacea (Prevotella spp.), Bacteroidaceae (Bacteroides spp.). Firmicutes belonging to the families Clostridiaceae and Ruminococcaceae (Fecalibacterium spp.).

Flow cytometry in the study of the subpopulation composition of T regulatory (Treg) lymphocytes in patients with IBS revealed an increase in the number of CD45R0+CD62L+ central memory cells (CM), which can regulate the processes of maturation and differentiation of lymphocytes in lymphoid tissue. A decrease in the expression of exonucleases CD39 and CD73 was detected, which can have a significant effect on their activity. A reduction in effector memory cells (EM) Treg was observed.

Changes in the expression level of exonucleases CD39 and CD73 were inversely correlated with the content of Proteobacteria and the representation of the genera Bifidobacterium spp. and Faecalibacterium spp. The content of СЫ Treg was directly correlated with the content of Dorea spp.

The results may be indicative of impairment in the processes of Treg differentiation, which are closely related to changes in key components of intestinal microbiocenosis in IBS.

The aim of this work was to reveal the interrelations between the number of T regulatory cells (Tregs) in patients with kidney cancer (KC) and phenotype of peripheral blood monocytes and their capacities to produce ROS. Patients with KC (T3N0M0, clear cell type) were examined prior to surgical treatment. Tregs phenotype and blood monocytes were identified by flow cytometry. ROS production of purified monocytes was carried out through the determination of lucigenin- and luminol-dependent spontaneous and zymosan-induced chemiluminescence activity. It has been found that the relative number of Tregs within total lymphocyte subset in KC patients was increased if compared to control values (in KC patients — Me = 6.3%). Then the patients were divided into two groups according to the median of Tregs number (less and more than 6.3%). The most pronounced changes in the phenotype of monocytes and their chemiluminescent activity were found in KC patients with the Tregs count of less than 6.3%. Our findings suggest that low frequency of Tregs in the periphery was associated with increased relative numbers of “intermediate” and “non-classical” (“pro-inflammatory”) monocytes as it was shown on the samples from patients with KC with a low level of Tregs. According to our data, both groups of KC patients had low levels of HLA-DR expression when comparing to control group. Furthermore, both groups of patients had decreased rates of HLA-DR and CD64 co-expressing cells. Changes in the phenotype of monocytes in patients with KC were closely linked with imbalance in ROS production. Thus, the monocytes spontaneous superoxide radical (primary ROS) synthesis in KC patients with a low Treg numbers were characterized by redused NADPH-oxidase activation time and increased level of its activity if compared to patients with a high Treg rates in peripheral blood. Next, the activation index for lucigenin-dependent chemiluminescence in KC patients was reduced, as well as it was independent of circulating Tregs rates and was determined apparently by the insufficiency of metabolic reserves. Similarly, spontaneous secondary ROS production by the monocytes in KC patients was lower then in healthy controls and was also independent of circulating Tregs rates. Finally, the induced secondary ROS synthesis and activation index for their synthesis in monocytes were reduced only in patients with KC with a low number of Tregs in the blood. In general, the characteristics of the chemiluminescent reaction of monocytes in patients with KC determined the imbalance in peripheral blood monocytes primary and secondary ROS production. Monocytes in patients with KC with a low number of Tregs in the blood were characterized by more pro-inflammatory activity due to the rapid activation and intensity of the synthesis of primary ROS.

Development of vaccines with a broad-spectrum of protection is one of the priorities in the programs of influenza prevention. Recently, the conserved fragments of influenza virus proteins (M1, M2, NP, the second subunit of the hemagglutinin HA2) provoke interest of investigators as the object of the development a broad-spectrum vaccines. Low immunogenicity present a problem when developing vaccines based on such conserved fragments. However, fusion of low immunogenic antigens into the high immunogenic carrier protein may significantly enhance their immunogenicity. The candidate vaccine protein Flg-HA2-2-4M2e was developed which containins two highly conserved viral antigens (the ectodomain of the M2 protein (M2e), 76130 region of the second subunit of HA2), fused with flagellin as a carrier protein. Flagellin (bacterial flagella protein) is a natural ligand of TLR-5, and has a strong adjuvant activity at different ways of its administration. The purpose of this study was to assess development of humoral and T cell immune response, along with broad-spectrum protection after mice immunization with the candidate Flg-HA2-2-4M2e vaccine protein. Mice were immunized intranasally three times with two-week intervals. Two weeks after the final immunization, the mice were challenged at the 5 LD₅₀ dose with influenza viruses A/California/07/09 (H1N1) pdm09 (phylogenetic group I), or A/Shanghai/2/2013 (H7N9) (phylogenetic group II). The results obtained in this study showed induction of strong M2e-specific humoral response (serum IgG and A) in the immunized mice. Immunization with recombinant protein stimulated formation of M2e-specific and virus-specific CD4⁺ and CD8⁺T cells in lung which produced TNFα or IFNγ. Production of antigen-specific effector and central memory T cells was also detected in lungs of immunized mice. The formation of cross-protective immunity in immunized mice was demonstrated in a model of lethal influenza infection. The experimental animals were almost completely protected from the high dose of the pandemic virus A/H1N1pdm09, and highly pathogenic avian influenza A/H7N9 (90-100% survival). We also evaluated the changes of antigen-specific immune response in immunized mice after sublethal infection with A/H3N2 influenza virus. Mice of control and experimental groups were infected with MID₁₀₀ of influenza virus A/Aichi/2/68 (H3N2). It was shown that the M2e-specific response (IgG, IgA) was significantly increased in immunized mice after sublethal infection with influenza virus A/H3N2, and we detected the changes in profile of M2e-specific IgG subclasses. Following sublethal infection in immunized mice, the proportion of M2e-specific IgG2a was increased 10-fold. The results showed that the recombinant protein Flg-HA2-2-4M2e is a promising candidate for development of universal vaccines, which induces a protective humoral and T-cell response to conserved viral epitopes and protects against influenza A viruses of both phylogenetic groups.

The problems of oncological disease treatment are considered relevant and timely issues of the current research programs. Since monotherapy is increasingly clear to be less effective than combination therapy, the novel studies seek for advancement of current treatments and development of new ones employing oncolytic immunotherapy being among the most rapidly evolving approaches. Modern genetic engineering techniques enable new applications of oncolytic viruses in the frames of combined cancer therapy. These applications are feasible, due to the abilities of oncolytic viruses to destruct tumor cells, like as by changing susceptibility of cancer cells to anti-tumor drug, and upon the whole body, thus overcoming the mechanisms conferring immunoresistance of tumor cells. In the present work, we have developed a recombinant vaccinia virus which is a promising platform for designing the antitumor drugs. The following modifications of viral genome were made by means of genetic engineering: gene encoding granulocyte-macrophage colony-stimulating factor was inserted into the region of viral thymidine kinase gene; viral A34R gene encoding a membrane glycoprotein, was replaced by A34R gene with two nucleotide substitutions resulting into D110N and K151E mutations which cause increased proportion of extracellular enveloped virions during the virus reproduction. Some properties of the recombinant virus were studied in vitro. The virus was shown to produce granulocyte-macrophage colony stimulating factor, and high numbers of extracellular enveloped virions. The genome modifications had no effect upon viral replication.

Aclinical case ofprimary immunodeficiency state (PIDS) is described: X-linked agammaglobulinemia in the boy of 8 years old. The results of molecular genetic studies: gene btk (ex 1-19) genome version GRCh38.p5, transcript version ENST00000308731 single nucleotide substitution identified с.1027С > Т in homozygous state leading to premature stop codon p.Gln343Ter (p.Q343*). The presented clinical case reflects the low alertness of pediatricians for PIDS. At the same time, a delay in diagnosis and prescription of treatment aggravates the prognosis of the disease course and reduces the quality of the patient life.

Eosinophilic granulomatosis with polyangiitis (EGPA), previously called Churg—Strauss syndrome, represents a rare form of ANCA-associated necrotising vasculitis which affects small vessels. This disease is characterized by typical combination of immunological disturbances, hypereosinophilia, severe bronchial asthma, transient pulmonary infiltrates, and kidney injury which is less frequent than in other forms of necrotising vasculitis.

Verification of the diagnosis is often hampered by comorbidities, such as chronic obstructive pulmonary disease (COPD) in the patients with long-term smoking history and occupational hazards.

In this article, we report a clinical case of EGPA in elderly patient with preceding COPD which caused diagnostic difficulties for this eosinophilic syndrome. Clinical pattern at the beginning of disease was presented by moderate inspiratory dyspnea and cough with small amounts of mucus sputum, which appeared after longterm exposure to chlorine-containing substances. COPD diagnosis in this patient was based on clinical pattern, long smoking experience, and occupational hazards. However, persistence of the symptoms during the ongoing therapy, as well as multidirectional dynamics of transient pulmonary infiltrates found on repeated CT-scans, prompted us to intensify diagnostic search for a systemic disease. Clinical, laboratory and instrumental signs of bronchial asthma were revealed, as well as hypereosinophilia and sensory polyneuropathy, which, if combined with CT-scan data, allowed us to prove the EGPA diagnosis.

This case shows that, despite great value of immune diagnostics, with negative blood tests for ANCA, it is necessary to detect mutually complicating comorbid pathology. EGPA was considered the basic diagnosis, and COPD as accompanying disorder, taking into account such reasons as an unfavorable prognosis for EGPA and the need for long-term chemotherapy with systemic corticosteroids and monoclonal antibodies. ANCA-negative testing in the patient, absence of severe kidney and skin lesions allows to suggest better clinical prognosis in this patient.