Molecular genetic mechanisms, signaling pathways, cultural conditions, factors, and markers of osteogenic differentiation of mesenchymal stem cells (MSC) are actively studied despite numerous works in this area of cellular technologies. This is largely due to the accumulating contradictions in seemingly classical knowledge, as well as permanent updating of the results in the field. In this regard, we focused on the main classical concepts and some new factors and mechanisms that have a noticeable regulatory effect on the differentiation potential of postnatal MSCs. The present review considers the significance of MSC sources for their differentiation capacity, as well as the role of the cellular microenvironment. The issues of classification, terminology, and functional activity of MSCs from various sources are discussed. The paracrine potential of MSCs in tissue regeneration has been considered; sufficient importance of inflammation in osteogenesis is noted, in particular, the presence of inflammatory cytokines and chemokines in the lesion focus, produced not only by microenvironmental cells but also by blood cells, including mononuclear leukocytes, migrating to the affected site. An important role in this review is given to biomechanical signals and to influence of conformational changes in cell cytoskeleton (cell shape) upon MSC differentiation, since the morphological features of cells and the structure of cytoskeleton are modulated by interactions of the cell surface with environmental factors, including hydrostatic pressure, fluid flow, compression/stretching loads. The data are presented concerning elasticity of extracellular matrix being a determining factor of cell differentiation. We conclude that one should switch from point studies of individual gene effects to multiple measurements of the gene-regulatory profile and biomolecules responsible for multiple, still poorly studied osteogenic factors of endogenous and exogenous origin. Among cornerstones in future (epi)genetic studies will be to decide if osteomodulatory effects are realized through specific signaling pathways and/or via cross-signaling with known genes controlling osteogenic differentiation of MSCs. 

Natural killer cells (NK cells) are cytotoxic lymphocytes that play a pivotal role in maintaining immunological surveillance and in developing an innate immune response. Since the discovery of NK cells in 1973, the mechanisms of their functioning have been studied in details, and there is currently no doubt that they play a special role in the process of recognition and destruction of transformed and malignant cells. Understanding the role of NK cells in antitumor immunity, on the one hand, leads to emergence of new immunotherapeutic strategies and, on the other hand, allows to adjust the existing treatment regimens for tumor diseases, in accordance with the principle of primum non nocere. Optimization of cancer therapy protocols executed in order to protect immune cells from death and functional impairment is an important problem that cannot be successfully resolved without regular aggregation of the results from disparate studies and critical analysis of the all accumulated data.

The objective of this review is to create a relevant and holistic picture of changes in the phenotypic and functional characteristics of NK cells in patients with two related hematological diseases – myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). For the treatment of both illnesses, drugs from the group of hypomethylating agents are used, the acting mechanism of which, unlike classical cytostatic agents, is based on modulation of the tumor cell genes expression. All the cells of the body are being affected, including NK cells, since these drugs act nonspecifically. Such an interaction leads to a hypomethylation of NK cell DNA and changes the expression of functional receptors, which, in turn, provide the development of antitumor NK cell immune response.

Of course, just the fact of changing gene expression in certain cells does not allow us to fully judge the drug’s impact on the state of immune system. Meanwhile, the origin of this change and its role are important in the context of the disease pathogenesis. Ultimately, a simple description of an increase or decrease in a single receptor expression is not illustrative, since it can lead to uncertain consequences. For this reason, the current review, in addition to describing the existing data on the changes of NK cell receptors expression under the influence of hypomethylating drugs, gives a special attention to critical analysis of functional characteristics of NK cells, including their cytotoxic activity aimed at malignant blast cells, being a determinant of clinical course in the described diseases. 

Chronic obstructive pulmonary disease (COPD) is a progressive disease characterized by irreversible or partially reversible obstruction of the bronchial tree. Currently, there are many proven links in the COPD etiopathogenesis, among which a pivotal role is assigned to the value of the hyperergic inflammatory reaction in response to inhalation of various harmful substances (tobacco smoke, industrial pollutants, etc.). The number of macrophages, neutrophils, lymphocytes increases in the lungs of COPD patients, and these cells secrete a fairly wide range of inflammatory mediators. Bacterial colonization of the airways is one of the key features in COPD pathogenesis leading to persistent or chronic stimulation of immune cells through Tolllike receptors (TLR), which perceive the pathogen-associated molecular patterns (PAMPs).

This article provides a review of literature concerning modern concepts of the role of Toll-like receptors expression and polymorphism, in particular, TLR4, in pathogenesis of COPD. TLR4 is a member of the Tolllike receptor family that plays a fundamental role in pathogen identification and innate immune activation. By recognizing the pathogen-associated molecular patterns (PAMPs) expressed on infectious agents, TLRs mediate the production of cytokines necessary for the development of effective immunity. Different TLRs exhibit distinct expression patterns. This receptor is most abundantly expressed in placenta and in the myelomonocytic leukocyte subpopulations. E.g., Di Stefano A. et al. (2017), determined immunohistochemically the expression levels of TLR2, TLR4, TLR9, NOD1, NOD2, CD14, Toll-interleukin-1-receptor domain containing adapter protein (TIRAP) and interleukin-1-receptor-associated phosphokinases (IRAK1 and IRAK4) in bronchial mucosa of patients with stable COPD of varying severity. It was found that TLR4 expression of the bronchial epithelium positively correlated with degree of obstruction and CD4+ and CD8+T cell contents. Stimulation of TLR4 increases cytokine production, which may be a relevant mechanism by which bacteria cause excessive inflammation in COPD patients. The degree of TLR4 involvement into COPD pathogenesis requires more detailed study in future, in order to determine the main mechanisms for emerging inflammatory response in the airways. This review article is part of a research grant project to study pro-inflammatory response to endotoxin of Gram-negative flora in COPD pathogenesis (State registration number – АААА-А19-119122390040-2).

Atopic dermatitis is one of the most common chronic inflammatory skin diseases caused by both terminal defects in keratinocyte differentiation, and pronounced type 2 immune responses. Atopic dermatitis is a fairly heterogenous disease, depending on the age subtype caused by activation of the Th22, Th17/IL-23 and Th1 cytokine pathway. Clinical studies using classical and targeted therapies have helped to determine contribution of various immune axes to the disease phenotype.

We present the modern activation theory mediated by Th2 reactions, due to congenital lymphoid cells of the 2nd group. Correlations between immune response in acute (IL-4, IL-5, IL-13, IL-31, CCL18, IL-22, S100A proteins) and chronic (IFNγ, CXCL9, and CXCL10) manifestations of atopic dermatitis are described. The theory of relationship between clinical manifestations and overexpression of some cytokines (IL-4, IL-13) is discussed. The correlation was shown between peripheral blood phenotype in atopic dermatitis of early childhood and in adult patients and individual production of serum biomarkers. In addition to excess Th17 production, early onset of atopic dermatitis in children correlated with elevated levels of antimicrobial peptides, which may serve as a signaling marker that triggers the disease. The article provides information about relationship between atopic dermatitis and other systemic non-allergic processes and diseases (psoriasis, atherosclerosis, cardiovascular diseases, obesity). Despite different polarity of T cells in atopic dermatitis and psoriasis, and different groups of cytokines produced in these diseases. Psoriasis is most of all due to Th17 associated with activation of IL-17, whereas atopic dermatitis is a consequence of Th2 dominance and associated excessive production of IL-4 and IL-13. The both diseases show activation of Th1 and Th22 with increased production of interferon-γ and IL-22, respectively. The article also concerns an interesting hypothesis on effects of the TWEAK protein upon clinical course of atopic dermatitis and psoriasis. In response to increased TWEAK activity, keratinocytes and skin fibroblasts produce a number of chemoattractant and pro-inflammatory factors commonly found in atopic dermatitis and psoriasis, in particular IL-13 and IL-17. TWEAK is not a single etiological factor for atopic dermatitis or psoriasis, but it causes the production of chemokines that promote chemotaxis of pathogenic inflammatory cells into the skin. With further studies of this pathogenetic factor, it will be possible to synthesize a new targeted drug for the treatment of atopic dermatitis and psoriasis. 

IgA is an important component of the mucosal system of the body. It limits penetration of pathogens into the bloodstream. Inflammatory diseases such as Crohn disease and colitis may be associated with disorders of IgA synthesis. Both B1 and B2 cells are a source of IgA in the intestines. Special attention is paid to B1 cells, which are able to respond to T-independent type 2 antigens and produce natural antibodies. B1 cells produce about 50% of the intestinal IgA including specific antibodies to the components of microorganisms contained in the gastrointestinal tract. The mechanism of IgA formation in the T-independent way is not investigated in details. It was suggested that the γδТ-cells promote switching to IgA synthesis by B1 cells. This assumption may be supported by their co-localization with B1 lymphocytes in the intestinal mucosa, as well as participation, along with B1 cells, in formation of the first-line defense against the pathogens. In addition, the both lymphocyte subpopulations evolve during initial ontogenesis, earlier than “classic” В2 and αβT cells. Therefore, it was suggested that γδT lymphocytes may be involved into the processes of induction and/or regulation of IgM and IgA production by B1 cells in response to TH2 antigens.

In the present study, we have shown the effect of γδT cells upon generation of IgM- and IgA-forming B1 cells in response to α-1,3-dextran in vitro. We also studied the dynamics of the mRNA expression for IgM- and IgA-heavy chains by the B1 cells at different terms of in vitro culture.

It was found that, during co-cultivation of B1 cells with 20% γδT lymphocytes, there is no increase in the number of dextran-specific IgM-producing cells. The B1 cells exhibited an increase of IgM heavy chain mRNA expression in response to dextran but not in co-cultures. Expression of mRNA for IgM heavy chains in co-cultures was decreased compared to non-treated B-cell cultures. Contrary to the earlier assumption, a presence of γδT lymphocytes in culture did not enhance the formation of IgA producents. The obtained data suggest regulatory properties of the γδТ lymphocytes during the B1 cells response to T-independent antigens. 

This article is the second communication in a series of articles devoted to the effects of a domestic preparation of macrophage-activating factor (GcMAF-RF) and assessment of its biological properties. The aim of this work was to study the effect of the GcMAF-RF upon M0 → M1 polarization of macrophages (Mph), and activation of the professional properties of ex vivo generated antigen-presenting dendritic cells (DC), as well as on ex vivo production of pro-inflammatory (TNFα, IL-1β, IL-6, IFNγ, IL-17, IL-18) and anti-inflammatory (TGF-β, IL-4, IL-10) cytokines, growth factors (IL-2, GM-CSF, G-CSF, VEGF) and chemokines (MCP, IL-8) by the whole blood cells from healthy donors. Mph and DC were generated from the monocytes (3 to 5×106 /ml) derived from adherent fraction of peripheral blood mononuclear cells (MNC) of healthy donors. Granulocyte/macrophage colony-stimulating factor (rhGM-CSF) was used to obtain Mph, whereas DC production was induced by GM-CSF and interferon-α. To provide M1 polarizing signals, bacterial lipopolysaccharide (LPS from E. coli 0114:B4) was used in controls. In experimental series, GcMAF-RF was added 48 h before the end of culture. The stimulating effect of the obtained Mph and DC upon cell proliferation was assessed in allogeneic mixed culture of leukocytes (alloMLC) using radiometric technique, by 3 H-thymidine incorporation. The influence index (IR) of Mph or DC upon allo-SCL was calculated as the ratio of the proliferative response of MNCs in the presence of Mph, or DC to the level of spontaneous MNC proliferation. To determine the cytokine production by human whole blood cells ex vivo, peripheral blood samples from 3 donors with two replicate GcMAF-RF preparations were used, at a total of 6 points. All variants of the study were carried out with mitogen-activated and non-activated blood cells. The cytokine content was determined by the ELISA assays. The effects of GcMAF-RF were quantified as a fold increase (FI), i.e., the ratio of cytokine production in the presence of GcMAF-RF to the level of their spontaneous production. It was shown that the GcMAF-RF preparation was as effective, as lipopolysaccharide (LPS), the standard Mph and DC activator which induces polarization of differentiated M0-macrophages into M1 cells and final maturation of DCs, manifesting by a significant increase in their allo-stimulatory activity in a mixed leukocyte culture (allo-MLC). Moreover, GcMAF-RF stimulates production of numerous cytokines and chemokines (TNFα, IL-1β, IL-6, IL-18, IL-4, IL-10, GM-CSF, G-CSF, VEGF, IL-8), by blood cells (granulocytes, lymphocytes, monocytes), thus indicating direct participation of the macrophage activator GcMAF-RF in various immune processes. The domestic GcMAF-RF drug induces polarization of macrophages M0 → M1, final maturation of DCs and allostimulating activity of Mf and DCs, and is also able to effectively stimulate circulating blood cells to synthesize cytokines/chemokines with pro-inflammatory and immunoregulatory activities. 

Extracellular vesicles that are shed from the plasma membranes take an active part in intercellular communication, transporting a wide range of molecules, including proteins, lipids, nucleic acids and carbohydrates, being of great functional importance. One of the steps to better understanding of distant communications of cells and their regulatory mechanisms is a proteomic study of various extracellular vesicles, including microvesicles and exosomes. Pro-inflammatory cytokines produced by monocytes and individual complement system components play a key role in their specific functioning. The aim of this work was to study proteomic composition of THP-1 monocyte-like cells and their microvesicles. The MALDI-mass spectrometric analysis of electrophoretic protein fractions of cell lysates and microvesicles allowed for identifying 107 proteins that perform various functions. Among 19 determined functional groups, the largest ones comprise transcription regulators and proteins with unknown functions. The smallest functional groups include regulators of cell differentiation and development, proteins participating in immune response and inflammation, cellular receptors and their regulators, transporter and transport regulatory proteins, as well as cell proteins mediating adhesion and matrix structures, processing regulators, proteins of ubiquitin-proteasome system, intracellular signaling, autophagy and exocytosis regulators, chromatin structural proteins, hemostatic regulators, and peptide hormones. An intermediate position is occupied by cytokines and growth factors, enzymes, cytoskeleton and motor proteins, as well as RNA processing and translation regulators. The subsequent DAVID Functional Annotation Clustering analysis allowed for identifying the most common groups distributed by their molecular function, biological processes, and cellular component. Separately, in the microvesicles derived from THP-1 monocyte-like cells, proteins of the immune response and inflammation, cytokines and growth factors, intracellular signaling proteins, cell differentiation regulators and developmental proteins, as well as cell adhesion and matrix proteins were identified among other protein molecules. The data obtained on the partial proteome of THP-1 monocyte-like cells and their microvesicles extend the existing knowledge on distant communications between the cells and suggest new mechanisms of interaction between monocytes/macrophages and their microenvironment. 

Determination of inflammatory markers in blood of conventionally healthy people is of interest due to opportunity of detecting diseases at early (preclinical) stages, as well as latent forms of pathological processes. The level of inflammation may serve as an additional criterion to forming control groups in clinical and biological studies. The aim of the study is to identify some inflammatory and autoimmune markers in a group of conventionally healthy people and to conduct a cluster analysis of the data obtained. The study involved 100 apparently healthy people (without clinical signs of infections, somatic, neurological or mental diseases) aged 19 to 88 years. The levels of IL-10, TNFα, IL-6 and autoantibodies to S100b and MBP were determined in blood serum using ELISA. Enzymatic activity of leukocyte elastase (LE) and functional activity of the α1-proteinase inhibitor (α1-PI) were determined spectrophotometrically. Protease inhibitory index (PII) was calculated as the ratio of LE to α1-PI. Cluster analysis, as well as the Shapiro–Wilk, Kruskal–Wallis, and ANOVA methods were used as the main approach to statistical data processing. All the subjects were divided into three clusters, according to their immunological parameters. The selected clusters were statistically significantly different from each other, in terms of LE activity, protease-inhibitory index (PII), as well as IL-10 and TNFα levels. The indices of a distinct cluster (43% of total cohort) are most close to average indices assessed for the general sample, which gives ground to consider the values of immune indicators from this cluster as physiological norm, corresponding to the background immunity state in healthy people. Combination of immunological parameters in two other clusters (30 and 27% of the subjects, respectively) may reflect different variants of inflammatory reactions. These clusters are characterized by multidirectional changes in LE activity and protease-inhibitory index, compared to the standard values, thus suggestive for different variants of latent inflammatory reactivity, which are realized in the patients presented in these clusters. The obtained clusters did not differ by age of the subjects (p = 0.3476), which makes it possible to exclude a significant influence of age on the determined immune parameters, and by gender characteristics (p = 0.7233). The selected clusters did not differ statistically in the functional activity of α1-PI and in the level of autoantibodies to S100b and MBP.
Thus, the group of conditionally healthy people is heterogeneous in terms of inflammation markers. Inflammatory reactions of varying severity were detected in about half of the cases. Probably, this may indicate the presence of a latent pathological process and requires a detailed clinical examination. 

Metabolic syndrome (MS) is among the main public health challenges worldwide, leading to significant labor losses, increased costs for treatment and rehabilitation of the patients. The aim of the present study was to identify the informative serum interleukins, by determining the odds ratio in elderly patients with MS and hypertension. The main group of 86 patients with MS and arterial hypertension (AH) aged 60-75 years was examined under clinical conditions. The inclusion criteria were as follows: age of 60-75 years, presence of MS, primary hypertension (grade II-III), absence of acute myocardial infarction, malignant neoplasms, disorders of cerebral circulation, kidney failure over last 6 months. Diagnostics of MS and hypertension was carried out in accordance with Expert Guidelines from the Russian Research Society of Cardiology on the MS Diagnosis and Treatment. Our first study of a large range of serum interleukins in elderly patients with MS and hypertension allowed us to reveal the inversely directed changes in pro- and anti-inflammatory cytokine contents. Combined AH/MS in elderly persons is accomplished by sufficient increase of the most proinflammatory cytokines, and vice versa, by significant decrease in anti-inflammatory cytokines in blood serum. This finding clearly points to importance of immunological regulatory systems for initiation of AH with MS at older age. Pro- and anti-inflammatory serum interleukins are actively involved into the AH/MS development in elderly accompanied by their pronounced imbalance. The mentioned immune reactions could underlie the MS/AH condition. High risk of this disorder is connected with changed production of proinflammatory cytokines (IL-8, IL-1β), like as anti-inflammatory serum interleukins (IL-4, IL-10), with predominance of the former. The above interleukins should be considered dominant diagnostic markers of AH/MS in elderly persons. Measurement of serum interleukins and discriminant-based approach allows highly reliable differentiation of elderly patients with AH/MS from similar individuals without this disorder. 

COVID-19, an infection caused by the new coronavirus SARS-CoV-2, is associated with a number of pathophysiological mechanisms, mobilizing a wide spectrum of biomolecules, mainly, cytokines.

The purpose of this study was to evaluate levels of multiple cytokines in blood plasma from the patients with COVID-19 during acute phase of the disease, and upon complete recovery. Samples of peripheral blood plasma of 56 patients with COVID-19, 69 convalescents and 10 healthy individuals were examined. Concentrations of 46 molecules, such as IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A/CTLA8, IL-17-E/IL-25, IL-17F, IL-18, IL-22, IL-27, IFNα2, IFNγ, TNFα, TNFβ/ Lymphotoxin-α (LTA), CCL2/MCP-1, CCL3/MIP-1α, CCL4/MIP-1β, CCL7/MCP-3, CCL11/Eotaxin, CCL22/MDC, CXCL1/GROα, CXCL8/IL-8, CXCL9/MIG, CXCL10/IP-10, CX3CL1/Fractalkine, IL-1ra, IL-10, EGF, FGF-2/FGF-basic, Flt3 Ligand, G-CSF, M-CSF, GM-CSF, PDGF-AA, PDGF-AB/ BB, TGF-α, VEGF-A were measured via xMAP multiplexing technology. Significantly increased levels of 18 cytokines were found in blood plasma from COVID-19 patients during acute phase of the disease (as compared to control group), i.e., IL-6, IL-7, IL-15, IL-27, TNFα, TNFβ/Lymphotoxin-α (LTA), CCL2/MCP-1, CCL7/MCP-3, CXCL1/GROα, CXCL8/IL-8, CXCL10/IP-10, CXCL9/MIG, IL-1rа, IL-10, M-CSF, GM-CSF, VEGF-A. We found a significant decrease of nearly all the mentioned cytokines in recovered patients, in comparison with those who had moderate, severe/extremely severe disease. Moreover, we revealed a significantly decreased level of 8 cytokines in plasma from convalescents, as compared with control group, i.e., IL-1α, IL-2, IL-9, IL-12 p40, IL-18, CCL22/MDC, Flt3 Ligand, TGF-α. Immune response caused by SARS-CoV-2 infection involves multiple cytokines, mostly, with pro-inflammatory effects. We have shown for the first time that the convalescence phase is characterized by significantly lower levels of cytokines which regulate cellular differentiation and hematopoiesis (in particular, lymphocytes, T-cells and NK-cells). Over acute phase of the disease, the levels of these cytokines did not change. We revealed a significant decrease of most plasma cytokines upon recovery as compared to acute phase. On the contrary, acute phase of the disease is accompanied by significant increase of both pro- and antiinflammatory cytokines in blood plasma. 

Expansion of myeloid-derived suppressor cells (MDSCs) due to impaired differentiation of myeloid progenitor cells under conditions of inflammation was described in a number of autoimmune diseases, including rheumatoid arthritis, systemic lupus erythematosus, and type 1 diabetes mellitus. Studying the role of MDSCs in ankylosing spondylitis is an important issue, given that increased concentration of proinflammatory mediators in this pathology can also cause myelopoiesis disorders. The aim of present work was to study the quantitative content of MDSC subpopulations in patients with different clinical phenotypes and activity of AS. 37 patients, including 10 patients without peripheral skeletal lesions (axial form) and 27 patients with simultaneous lesions of spine and peripheral joints (peripheral form) were recruited into the study. The control group consisted of 32 age/sex-related healthy donors. Evaluation of granulocytic (LinHLA-DRCD33+CD66b+; G-MDSC), monocytic (CD14+HLA-DRlow/-; M-MDSC) and early-stage MDSCs (LinHLA-DRCD33+CD66b- ; E-MDSC) was performed using corresponding antibodies (BD Biosciences, USA) in the population of peripheral blood mononuclear cells by flow cytometry. In general, the AS patients were characterized by an increased relative and absolute amount of M-MDSC (p = 0.00002 and p = 0.00003, respectively) and G-MDSC (p = 0.0002 and p = 0.0006, respectively). Patient gender, age, and HLA-B27 expression did not significantly affect the content of these cells in peripheral blood. An increase in the median values of M-MDSC was detected both in patients with axial (Ме 5.0 (3.2-6.3) versus 2.4 (1.7-3.5) %; p = 0.001) and peripheral form (Ме 5.0 (3.0-7.0) versus 2.4 (1.7-3.5) %; p = 0.0002) AS. At the same time, the G-MDSC expansion was observed only in patients with involvement of peripheral joints (Ме 0.16 (0.07-0.3) % versus 0.05 (0.04-0.09) %; p = 0.0001). The relative contents of E-MDSC, M-MDSC and G-MDSC in the axial form of AS was in direct correlation with the activity of the disease (R = 0.58, p = 0.02; R = 0.73, p = 0.08 and R = 0.65 p = 0.04, respectively). This relationship was not observed in peripheral form of AS. The data obtained suggest a potential involvement of MDSCs in pathogenesis and phenotypic heterogeneity of AS. Simultaneously, the revealed direct correlation between the MDSC contents and the disease activity suggests a decrease in suppressive activity and/or appearance of pro-inflammatory activity in MDSC, thus requiring further research in the field. 

Neutrophils play an important role in carcinogenesis, mediating inflammation, immunosuppression and metastasis. A dual role of neutrophils in regulation of angiogenesis has been shown. Endometrial cancer is the most common malignant neoplasm of the female reproductive system. To assess the cellular angiogenic potential, we determined the levels of inflammatory and angiogenic cytokines (VEGF-A, IL-17A, IL-1β, IL-6) in cell lysate of peripheral blood neutrophils in the patients with endometrial cancer, and with uterine myoma (comparison group). Expression of the nuclear factor-kB was determined. In nuclear fraction of neutrophils. The neutrophil-to-lymphocyte ratio (NLR) was assessed in the studied groups. Statistical processing of the obtained data was carried out using Statistica 10 software. We have not found any significant changes in NLR in endometrial cancer, compared with controls and the uterine myoma groups. Expression of NF-kB and VEGF was increased as compared to the control for all the studied stages of endometrial cancer and in uterine myoma. There was a change in NF-kB level in neutrophils, depending on the tumor differentiation grade. A regression relationship was found between the content of VEGF and NF-kB in neutrophils. We have found increased IL-1β and IL-6 levels in neutrophils of the uterine myoma patients, and at different stages of endometrial cancer compared with control. The IL-1β level was higher in neutrophils of the patients with intermediate and high tumor grade, compared to low-grade cases. IL-17A expression in the neutrophil lysate was significantly reduced in uterine myoma and at different stages of endometrial cancer, as compared with controls. There was a moderate inverse correlation between the contents of VEGF and IL-6 in neutrophils (r = -0.426, p = 0.001); a remarkable inverse relationship between VEGF and IL-17A (r = -0.615, p < 0.001). The combination of IL-6 and IL-17A levels in the neutrophils lysate (according to the results of multivariate analysis) may be used for the differential diagnosis of endometrial cancer and uterine myoma. Thus, the differences in the expression of inflammatory and angiogenic cytokines included in NF-kB-dependent signaling, may point to acquisition of pro-tumor functions by neutrophils during the endometrial cancer progression. 

Activation of cytokine network is one of the key mechanisms in immunopathogenesis of chronic periodontitis (CP), a common dental disease. Therefore, the study of the proteomic (including cytokine) profile of saliva is not coincidentally considered among global research areas in periodontology. However, the existing data on the contents of cytokines / chemokines in oral fluid (OF) in CP are contradictory. This ambiguity of the results can be associated both with a variety of methodological approaches to the cytokine determination, and with different severity of CP in the studied cohorts of patients. Moreover, recent systematic reviews and meta-analyses show that clinical value of chemokines in CP is also not yet determined thus indicating a need for future research in this area.

The aim of this study was to assess clinical and diagnostic value of some chemokines from oral fluid in chronic mild periodontitis. The work was based on a study of 18 patients diagnosed with mild chronic periodontitis (MCD-10 K05.3) and 12 practically healthy volunteers with relatively intact periodontal tissue. The diagnosis was based on standard clinical and radiological criteria. In all cases the levels of 8 chemokines were determined using multiparametric fluorescence analysis with magnetic microspheres (xMAP technology, Luminex 200, USA). Statistical analysis was carried out by methods of non-parametric statistics. To determine the predictive value of the test, ROC analysis was performed.

It has been shown that CP is accompanied by increased levels of CXCL1 (5.5-fold), CXCL8 (8.1-fold), CXCL12 (3.5-fold), CCL2 (2.7-fold). At the same time, the level of other chemokines did not change significantly. There was a lack of correlations between individual parameters in the group of patients with CP, in contrast to the control group, thus, probably reflecting disturbed mechanisms of “balance” regulation of chemokines. By means of ROC analysis, highly sensitive biomarkers of chronic mild periodontitis were identified. The diagnostic sensitivity and diagnostic specificity were for CXCL1 (91 and 75%, respectively), CXCL8 and CXCL12 (95 and 75%), CCL2 (82 and 75%). The data obtained indicate that the salivary chemokines CXCL1, CXCL8, CXCL10, CXCL12, CCL2 can be considered potential biomarkers of mild chronic periodontitis. 

In recent years, primary immunodeficiencies have turned from the class of rare diseases to the category of more common disorders which may be encountered by doctors of any clinical discipline. The first case of primary immunodeficiency disorder (PID) in Chuvashia was detected in 1993. Since that time, the Department of Internal Diseases with the Course of Clinical Immunology at the I. Ulyanov Chuvash State University registered all the cases of PID diagnosed in the region, introducing them into the Republican Registry of PID. The study was aimed for searching epidemiological indexes, clinical and laboratory manifestations of PID in Chuvash region. The study was based on the patient data obtained by retrospective analysis of 85 case histories of PID patients, treated at different departments of the Republican Clinical Hospital, and the City Chuvash Pediatric Clinical Hospital of Public Health Ministry in 2000-2019, as well as on 49 outpatient records of the patients included into the Regional PID Registry. Various forms of PIDs were diagnosed according to the criteria developed by the European Society for Immunodeficiency and the Pan-American Group on Immunodeficiency (1999). The results of this study showed that the incidence of PID in the Chuivash Region is 3.4:100,000. The incidence of common variable immune deficiency (CVID), the most common form of PID in the region, was 1.58 per 100,000 population. The average age at the time of CVID diagnosis in Chuvash patients was 30.4±16.1 years, and the age of CVID debut was 11.3±15.0 years. The delay in proper diagnosis from the moment of clinical manifestation of CVID was, on average, 17.9 years in the region. At the time of CVID diagnosis, the patients showed marked decrease in the levels of 3 or 2 immunoglobulin classes (IgG and IgA), and T-helper cell contents (CD3+CD4+) in peripheral blood. Prevalence of selective IgA deficiency with сlinical symptoms was 0.83 per 100,000 population of the region, and the incidence of the asymptomatic form of this PID was 1 : 167. In patients with selective IgA deficiency, there were also disorders in the T cell system manifesting as decreased relative number of cytotoxic T-cells as well as elevated IgG and IgM levels. The age of diagnosis of X-linked agammaglobulinemia in the region was 3.5±3.0 years. In addition to disturbances of humoral adaptive immunity in children with this disease, a decrease in absolute T cell numbers was detected. In conclusion, the article describes disturbances of postvaccinal immunity in a pregnant patient with CVID, with asymptomatic clinical course, thus leading to false interpretation of the serological markers of TORCH infections and wrong strategy of pregnancy management.

The relationship between the HLA-G gene polymorphism (rs41551813, rs12722477, rs41557518), intrauterine infection and recurrent miscarriage (RM) in women were studied. The case group consisted of 180 patients with RM, defined as two or more consecutive miscarriages (min = 2; max = 8) at up to 20 weeks of gestation, and with clinically confirmed pregnancies and non-viable fetuses. At the time of examination. the women were enrolled from the Genetic Counseling Center at the Kemerovo Regional Clinical Hospital, Kemerovo, Russia, and were not pregnant. Each patient underwent a gynecological examination. We excluded women with a history of medical abortion, birth, and ectopic pregnancies. In addition, we excluded women with endocrine (e.g. diabetes) disorders. To exclude other known causes of spontaneous abortion, the following tests were performed: ultrasound examination of pelvic organs, and karyotyping in women and men. The women’s mean age in the RM group, was 29.6±4.8 (SD) years. The control group comprised 408 fertile women. These women didn’t have a history of spontaneous abortion, or a family history of congenital malformations. They have born, at least, 1-2 healthy children. Women’s mean age at birth of last child was 26.8±5.2 (SD) years. Influence of the intrauterine infection was analyzed on the basis of laboratory tests. Diagnostics of bacterial vaginosis and vulvo-vaginal candidiasis by microscopic examination was conducted. Viral agent infections (herpes simplex virus type 2, cytomegalovirus, human papilloma virus type 16/18), Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, Gardnerella vaginalis and Trichomonas vaginalis were detected by enzyme-linked immunoassay and polymerase chain reaction (PCR). The data were obtained from the medical cards of the surveyed women. All the women gave a written informed consent before participating in the study. Typing of polymorphisms of Thr31Ser (rs41551813, HLA-G*01:03) in exon 2, Leu110Ile (rs12722477, HLA-G*01:04) and 1597 delС (rs41557518, HLA-G*01:05N) in exon 3 HLA-G genes were performed by realtime PCR followed by melting analysis. The study showed that the intrauterine infection was not a risk factor for RM (p = 0.30) in the examined women. It was found that the 110 Ile allele (HLA-G *01:04) was a risk factor for RM both in women with intrauterine infection [ORa = 4.50 (2.41-8.38), p = 2.09e-06], and in women without infection [ORa = 2.46 (1.44-4.21), p = 0.0009]. The cooperative influence of genetic and infections factors with the risk of RM in women was revealed [ORa+f = 3.50 (2.01-6.09), p = 8.78e-06]. Our results will be useful in understanding the molecular mechanisms of immune disorders in fetomaternal interface, and for choosing the strategy of management and treatment in women with RM. 

To treat the patients with oncological diseases, cellular immunotherapy have been actively developed in recent years. Usage of antitumor adoptive immunotherapy (AIT) based on in vitro activated lymphocytes and natural killer cells (NK cells) is the most urgent issue. Their antitumor potential is enhanced by in vitro cultivation in the presence of cytokines. In vitro incubation of autologous peripheral blood mononuclear cells supplemented with cytokines produces cytokine-induced killer cells (CIK). These cells represent the important component of antitumor protection which may be potentially used for AIT. Currently, the studies in the field of antitumor AIT continues, searching for high-tech and safe methods of cellular immunotherapy in oncology. The article describes results of in vitro activation of lymphocytes from cancer patients (n = 19) in the culture media containing IL-2, IL-12 and IL-15. The ELISA assays revealed differences in the cytokine secretion (IL-2, IL-4, IL-6, IL-10, IFNα, IFNγ, TNFα) after in vitro activation of lymphocytes. It was shown that, for earlier in vitro activation of cells, IL-15 should be supplemented. We also studied the ability of whole blood cells from cancer patients (n = 20) receiving cell immunotherapy vs those non-treated with AIT, as well as healthy donors (n = 10), to secrete cytokines in spontaneous and mitogen-induced cultures. Peripheral blood cells of cancer patients receiving AIT for a year or more proved to show a more pronounced ability to respond to mitogen stimulation, in contrast to the patients who have never received AIT, as well as in relation to a group of donors whose immune system has not been stimulated either. Most likely, this may be explained by the fact, that the in vitro activated CIKs if introduced into the patient’s body, trigger a number of cascadelike intercellular interactions leading to activation of the patient’s immune system cells, especially, antitumor immunity, proliferation of cytotoxic lymphocytes and NK cells, like as in vivo inhibition of immunosuppression. The developed method of CIK production may be offered for carrying out cellular AIT in oncological patients as a novel approach to in vivo activation of the patient’s immune system. 

According to WHO data, about 11 million people need medical care after burns every year. In the overall structure of burns, the share of thermal trauma (TT) is 80%. Lymphocytopenia in TT is a risk factor for infectious complications and limited repair, and the development of new tools for TT therapy using dermal films is demanded in combustiology. The aim of the study was to evaluate changes in blood lymphocyte parameters, i.e., quantitative composition and their death during experimental thermal damage under the influence of the originally developed dermal film with melatonin (MT) in 49 inbred rats. The grade IIIA TT of 3.5% body surface was modeled by contact with boiling water for 12 s. Dermal films based on sodium carboxymethylcellulose supplemented with MT at a concentration of 0.005 g/g were applied daily for 5 days. The total numbers of lymphocytes, CD45RA+ and CD3+ cells, counts of lymphocytes with signs of partial necrosis, early and late apoptosis were assessed in blood. Relative decrease in the area and rate of the burn wound epithelization were also calculated. In animals with TT, the number of blood lymphocytes decreased on days 5, 10 and 20, including CD45RA+ and CD3+, along with increased amounts of lymphocytes with signs of necrosis, late and early apoptosis. By the term of 20 days, the burn wound area was reduced by 11.5%. Usage of dermal films with MT increased the amount of CD3+ cells in blood on days 5 and 10, CD45RA+ on days 5, 10 and 20, being associated with decreased number of lymphocytes showing signs of early apoptosis on days 5, 10 and 20, as well as features of necrosis and late apoptosis on days 5 following TT, accelerates the healing of a burn wound on days 5, 10 and 20 after TT. with a 20 cent reduction of its area by the day 20. Epithelization rate of the burn wound when applying MT-supplemented dermal film on days 5, 10 and 20 increases, along with higher amounts of CD3+ in the blood, and reduced counts of lymphocytes with signs of early apoptosis. 

It is known that sufficient changes are observed in cellular and humoral links of immune system upon chronic exposure vapors of metallic mercury. In previous studies, upon development and in the course of the chronic mercury intoxication (CMI) we revealed pronounced regular changes of inflammatory mediators (IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, TNFα), and showed an important role of autoimmune reactions affecting nervous tissue proteins. Over last 20 years, an increased interest was shown for interleukin 17 (IL-17) and its role in a number of inflammatory and autoimmune diseases. However, there is no data on its role in neurointoxication with mercury. Considering that IL-17 has proinflammatory activity and stimulates production of the individual cytokines, the goal of our work at the next stage of research, was to identify quantitative changes of serum IL-17 in patients with mercury neurointoxication of various severity, aiming to substantiate additional criteria for early and effective diagnosis of the disease.

The study was performed in males chronically exposed to metallic mercury vapors with early signs of neurointoxication (n = 37), individuals diagnosed with CMI (n = 40), and “conditionally healthy” men (n = 34). Proper diagnosis confirmed by history of working contacts with a harmful industrial factor, and absence of comorbid pathologies served as inclusion criteria. Statistical processing of the results was carried out using the STATISTICA 6.0 application package (StatSoft, USA). The study has revealed a statistically significant increase in serum IL-17 concentrations, both in the patients with early signs of neurointoxication with metallic mercury vapors, and individuals with CMI, when compared with the comparison group, thus indicating its activation, and being consistent with results of several workers who showed an IL-17 increase in immunoinflammatory diseases. Correlation analysis has shown an association between IL-17 and inflammatory mediators, i.e., the patients with early signs of neurointoxication had an increased production of IL-17 accompanied by an increase in anti-inflammatory IL-10, whereas the CMI patients with an increase in IL-17 concentration showed a decrease in pro-inflammatory TNFα, thus confirming its role in immunopathogenesis of mercury neurointoxication. Further study of IL-17 involvement in the initiation and maintenance of chronic inflammation will not only contribute to better understanding of the disease origin, but also, most importantly, implication of novel, more effective treatments. 

Activated platelets aggregate with monocytes by binding membrane bound molecules. Platelet-monocyte interaction is considered to underlie pathophysiological mechanisms bridging thrombosis and inflammation. Detection and analysis of platelet-monocyte complexes (PMC) provide means for revealing their physiological and pathogenetic roles and are instrumental in the diagnostics of various pathological conditions including obstetric complications. The aim of the study was to develop the method of quantitative determination of peripheral blood PMC, that preserve phenotypic features of platelets and monocytes, and to reveal their changes by ex vivo analysis. The suggested procedure includes immediate fixation of blood sample, immunocytochemical staining with fluorochrome-conjugated specific antibodies against markers of activation and differentiation followed by lysis of erythrocytes, and flow cytometric analysis. Fourteen samples of peripheral blood from patients with history of pregnancy complication were obtained in first trimester of ongoing pregnancy and analyzed. It was demonstrated that quantitative and qualitative in vivo characteristics of PMC remained unchanged in fixed samples, whereas the number of PMC and expression levels of the markers of platelet and monocyte activation dramatically increased in the unfixed blood. The set of monoclonal antibodies and gating strategies, used in this study, ensure phenotyping and evaluation of percentage/absolute count of PMC in the total monocyte population (CD45⁺CD14⁺) and in the subpopulations of classical (CD14⁺CD16^-), intermediate (CD14⁺CD16⁺), and non-classical (CD14^lowCD16⁺) monocytes. This approach provides insight into the participation of different monocyte subsets in the formation of PMC and their roles in physiological and pathophysiological processes. In some samples, elevated PMC proportion was observed, accompanied by significant increase in the expression of platelet activation marker CD62P and decrease in the expression of its monocytic ligand CD162. These changes suggested altered activation of PMC and their participation in the pathophysiological mechanisms of some pregnancy complications. Immunophenotyping of PMC affords an opportunity to characterize their proinflammatory, procoagulant and adhesive properties; these results can be used for research and diagnostics. In particular, the method is suitable for detection and phenotyping of PMC in pregnancy complications and other pathological conditions associated with the disorders of hemostasis and thrombosis.