The ageing process should be considered with respect to all homeostatic systems of the body, their importance for the existence of the organism itself and different timing of their switching-on in the process of age-related changes. An attention should be paid to the literature data on age-related changes in the functional activity of immune system, which starts from almost zero at birth, reaches a maximum at the age of about 40-50 years and significantly decreases during the natural ageing process. The vast majority of most socially significant diseases of modern humans are reasonably associated with this last stage of age-related changes in the functional activity of the immune system. So far, there is probably no consensus on whether ageing is a disease, or not a disease, but just a kind of natural wear of the “biological machine”. In any case, one should emphasize that thymic gland is one of the first organs where negative age-related changes are revealed, one of the two central organs of the immune system. Bone marrow is the second central immune organ. Age-related changes in the thymus are not “all or nothing”, “now or never”, they are normally characterized by a gradual decrease of the cell production and changes in their qualitative characteristics, which undoubtedly result into some negative effects upon the indices of immune system activity. There is a need to develop new approaches towards the assessment of functional parameters of the thymus, highlighting the search for therapeutic approaches that would maintain functional activity of the immune system at a high level.

In the modern world, oncological diseases occupy the leading positions in the structure of mortality. An integrated approach to oncotherapy is not only aimed at immediate affection of malignant tumors, but also directed at reducing the risk of tumor recurrence and metastasis, as well as alleviating side effects of chemotherapy and radiotherapy of the disease. In oncologic disorders, blood viscosity increases, thus being associated with hypercoagulation syndrome. To prevent its consequences, the direct and indirect anticoagulants, especially heparin and its derivatives, are actively used. Biological functions and structural features of heparin make it a potential universal platform of a drug development for broad application, including oncology. With the advent of heparin fractionation technology and preparation of low-molecular weight forms and their derivatives, it has become possible to focus not only on anticoagulant activity but also to obtain fractions with targeted pharmacological activity. Usage of the anticoagulants has shown their antitumor activity in some cases, thus providing a basis for a more detailed study of pharmacotherapeutic effects of this group of drugs. Currently, some data suggest various pathways of interaction between heparin and tumor cells. There are multiple common features in development of a primary tumor and formation of secondary distant metastases, which may be attributed to similar molecular cellular mechanisms. The molecules mediating intercellular interactions, both between the tumor cells and between malignant cells and tumor-associated immune cells (e.g., lymphocytes and macrophages) may serve as targets for heparin thus helping the tumor to evade immune surveillance. The cytokines that stimulate tumor angiogenesis represent another important therapeutic target. Heparin derivatives are able to suppress tumor activity and prevent metastatic processes at various stages by inhibiting heparanase, P-/L-selectin, and angiogenesis activity, modulating the CXCL12-CXCR4 chemokine axis, and regulating OAM activity.

This brief review addresses the current understanding and application of the potentially antimetastatic properties of heparin and its derivatives in malignant bone tumors since the heparin-based drugs are used as anticoagulants in arthroplasty of large joints and bone defects in patients with osteosarcoma.

Monocyte-derived dendritic cells (DCs) can be used for cell immunotherapy of cancer. In most cases, mature DCs, loaded with tumor-associated antigens, are used for immune therapy. The functionality of DCs for immunotherapy substantially depends on their immunophenotype and secretory profile, which are established after DCs maturation. The purpose of this research was to explore the phenotype of DCs after using various approaches for stimulation of their maturation.

Maturation of DCs was stimulated by pro-inflammatory cytokines and their mixtures, or by ligands to the TLRs of DCs. DCs were stimulated by the following means: TNF; poly I:C; LPS; cytokine cocktail (TNF + IL-1 + IL-6 + PGE2); the cocktail mixed with poly I:C; and melanoma cells lysate. Forty-eight hours after stimulation, the expression of DCs’ receptors involved into their interaction with T cells, was evaluated by flow cytometry. Moreover, the secretion of IL-12 (activator of T cell response) and IL-10 (inhibitor of T cell response) was estimated by ELISA technique.

We have shown that, following stimulation with cytokine cocktail, the DCs exhibit highest expression of receptors, which are necessary for interaction with T cells and for activation of T cell mediated immune response, i.e., antigen-presenting receptors (HLA-DR), co-stimulatory receptors (CD83, CD40, CD86), and receptors controlling the migration of DCs to lymph nodes (CCR7). Moreover, the cocktail-stimulated DCs intensively secrete both IL-12 and IL-10. The stimulatory effect of TNF and poly I:C proved to be moderate: the expression of most receptors was significantly lower than after using the cocktail; no significant differences from control (in absence of induced maturation) in IL-12 secretion were detected. LPS and melanoma cell lysate did not affect both expression of receptors and secretory profile of DCs. Addition of poly I:C to the cytokine cocktail did not affect the receptor expression, but significantly increased the secretion of both proinflammatory IL-12 and anti-inflammatory IL-10.

The results of experiments demonstrate that the mixture of cytokine cocktail and poly I:C seems to be the most effective tool for stimulation of DCs maturation. However, further experiments are required to compare the functionality of DCs when using different tools for induced DC maturation.

Hypertrophy of pharyngeal lymphoid tissue is among the most common problems in pediatric otorhinolaryngology, in particular, hypertrophy of the palatine tonsils. This disorder is characterized by increased size of a single or both palatine tonsils combined with various clinical symptoms. The principles of treatment in children with this pathology remain debatable, since the long-term effects of bilateral tonsillotomy are still not fully understood. The aim of the study was to assess the expression levels of genes encoding the innate immunity molecules (TLR4, HBD1, HBD2, IL1β) in the mucous membranes of palatine tonsils in children with palatine tonsillitis before and after treatment.

We conducted a study of 78 patients divided into three independent groups. The 1^st group (comparison) included 20 somatically healthy children. The 2^nd group included 28 children with grade 2 palatine tonsil hypertrophy who underwent local treatment. The 3^rd group included 30 children with grade 3 hypertrophy of the palatine tonsils, who, by clinical indications, underwent bilateral tonsillotomy. Determination and evaluation of innate immunity indices was carried out both before starting the treatment, and one month later, using the real-time PCR method.

In children with hypertrophy of palatine tonsils, the initial values of TLR4 gene expression and antimicrobial peptides differed from those of healthy children. A decreased expression of HBD1 and HBD2 genes, which provide immediate protection against pathogens, was revealed. The values of TLR4 gene expression differed in groups of children with varying degrees of palatine tonsillar hypertrophy. In patients with bilateral tonsillotomy, an increased expression of TLR4 gene and a decreased expression of antimicrobial peptide genes (HBD1, HBD2) were revealed, which may indicate a readiness for development of tonsillar inflammation in response to pathogens. One month after surgical treatment, the indices of innate immunity were comparable with those of healthy children thus confirming the validity of surgical treatment. In the 2^nd group of patients, the TLR4 gene expression one month after conservative treatment remained reduced, the expression of β-defensin HBD1 gene increased and exceeded the indicators of the group of healthy children, the expression level of the IL-1β gene was reduced.

The revealed imbalance between the TLR4, HBD1 and HBD2 expression levels confirms an important role of innate immunity mechanisms in pathogenesis of palatine tonsillitis. The assessment of innate immunity indices may be used as an additional criterion in administration of therapy for hypertrophy of palatine tonsils and evaluation of its efficiency.

Atherosclerosis is accompanied by damage to the vascular endothelium of arteries followed by development of inflammatory response and formation of atherosclerotic plaques. Innate immunity is an important component of this response being the earliest non-specific key mechanism. Our objective was to perform a comprehensive assessment of the cellular link of innate immunity, and to compare the results obtained at various terms after coronary stenting.

The study involved 50 patients with coronary atherosclerosis (Group 1), who had clinical indications for stenting of coronary arteries, and 20 volunteers (Group 2), who have no signs of coronary artery disease. The study of immune parameters was carried out before surgery, at 4-5, 9-10 and 28-30 days after operation (during early postoperative period), as well as 6 and 12 months after stenting, i.e. over the late post-surgical period. Phenotyping of peripheral blood monocytes and lymphocytes was performed by flow cytometry using monoclonal antibodies (Beckman Coulter, USA). Intracellular content of Granzyme B was carried out with an FC500 flow laser cytofluorimeter. Metabolic activity of neutrophils was assessed by the NBT test. Alpha defensin was determined in blood plasma by ELISA technique (Hycult Biotech, USA). Statistical analysis was performed using the Statistica 12.0 program (StatSoft, USA). Statistical significance was considered significant at p ≤ 0.05.

The numbers of natural killer cells and their activity, as well as those of monocytes, were increased in patients with coronary atherosclerosis. We have also shown a suppression of antigen presentation processes, an imbalance in microbicidal activity of neutrophils, with predominant secretion of antimicrobial peptides. Over the early post-surgical period, significant changes included only decreased content of intracellular Granzyme B on days 4-5, and expression of TLR4 and HLA-DR on days 4-5 and 9-10. During the late period, the patients with coronary artery disease exhibited a significant decrease in the content of some lymphocyte subsets: CD3⁺CD16⁺, CD16⁺Gr⁺ as well as amounts of monocytes: CD14⁺CD282⁺, CD14⁺CD284⁺, CD14⁺CD289⁺, along with HBT-test activity and α-defensin contents, and increased numbers of HLA-DR-expressing monocytes.

There are changes in cellular component of innate immunity, indicating persistent inflammation in patients with coronary heart disease. The dynamics of revealed changes following coronary artery stenting may reflect a lability of assessed indicators mostly over the late postoperative period, thus serving a basis for predicting the outcome of coronary stenting.

Development of criteria for the optimal donor selection based on the analysis of the allo-HSCT results, high-resolution HLA typing for the donor and recipient resulted in decreased incidence of immunological complications, primarily an acute «graft-versus-host reaction». However, due to the pronounced allelic polymorphism of the main histocompatibility complex (MHC) genes, the search for an optimal donor is ineffective in a number of patients. To increase the chances of selecting a donor for patients with rare HLA genotypes, the hematopoietic stem cell donor registries recruit the persons of various nationalities. An increased number of donors from different ethnic groups provide a broader immunogenetic diversity of the donor cohort. Currently, the registry of hematopoietic stem cell donors at the Russian Research Institute of Hematology and Transfusiology includes representatives of 49 nationalities, most of which, are considered themselves Russians. The third largest ethnic group in the registry comprises Tatars. The purpose of this study is a comparative analysis of immunogenetic characteristics of potential hematopoietic stem cells donors in the registry, who have self-identified as Russians and Tatars. As a result of the study, we have not found significant differences in frequencies of HLA allelic groups in the compared cohorts, a trend for higher frequency of the HLA-B*27 group was noted in Tatars. However, significant differences have been revealed for the distribution of HLA haplotypes in Russians and Tatars. The most common HLA haplotype among Tatars was A*02-B*44-DRB1*07, being much less common in Russians (4.61% vs 0.55%, p = 0.002). HLA haplotype A*03-B*13-DRB1*07, belonging to the ten most common among Tatars, was significantly less frequently detected in Russians (1.62% vs 0.08%, p = 0.026). HLA haplotype A*03-B*08-DRB1*03 was also significantly more common in Tatars compared to Russians (1.42% vs 0.06%, p = 0.026). HLA haplotypes A*02-B*18-DRB1*11, A*02-B*15-DRB1*04, A*02-B*15-DRB1*13, presented in Russians at a frequency of > 1%, were not determined among the tested Tatars. HLA haplotypes A*31-B*58-DRB1*04, A*24-B*44-DRB1*01, presented in Tatars at a frequency of > 1%, were not detectable in Russians. The results of our study indicate a need for recruiting more representatives of the Tatar ethnicity to the registry, thus increasing immunogenetic diversity of the donor pool and resulting into increased chances of compatible unrelated donor selection for the patients with HLA haplotypes, which are now underrepresented in our registry.

HLA haplotype is a block of HLA genes located on the same chromosome. Highly polymorphic HLA genes display strong linkage disequilibrium, which results in conserved multilocus HLA haplotypes. Assessment of HLA haplotypic diversity of a specific population is important, particularly for allogeneic hematopoietic stem cell transplantation. Family pedigrees remain the gold standard for studying HLA haplotype segregation. HLA haplotypes, obtained by observations of the segregation of HLA alleles within the family, really exist in the human population. The aim of this work has been to establish the frequencies of HLA haplotypes A-B-C-DRB1-DRB3/DRB4/DRB5-DQA1-DQB1-DPA1-DPB1 in families of patients with assignment to HLA-typing for allogeneic hematopoietic stem cell transplantation. The study included 109 families of patients, in which patients and their potential relative donors of allogeneic hematopoietic stem cell were subjected to HLA-typing. Patients and members of their families were typed by the NGS method in the Laboratory of Tissue Typing at the National Medical Research Center for Hematology for 11 HLA genes – A, B, C, DRB1, DRB3, DRB4, DRB5, DQA1, DQB1, DPA1 and DPB1. The genotyping was performed by the NGS method using the AllType NGS 11 Loci Amplification Kits (One Lambda, USA) on the MiSeq sequencing platform (Illumina, USA). The sequences were analyzed using the TypeStream Visual Software (TSV) (One Lambda, USA) and the IPD-IMGT/HLA database 3.44. 360 copies of HLA-haplotypes were found in the studied families. The frequencies of HLA haplotypes were determined by direct counting. The most common 7-locus haplotype was A*01:01-B*08:01-C*07:01-DRB1*03:01-(DRB3*01:01-DQA1*05:01)-DQB1*02:01/163N, the most common 9-locus haplotype was A*03:01-B*07:02-C*07:02-DRB1*15:01-DRB5*01:01-DQA1*01:02-DQB1*06:02-DPA1*01:03-DPB1*04:01P. These HLA haplotypes (in brief, A-B-C-DRB1-DQB1) are the first and second most common HLA haplotypes in most Russian registries of bone marrow donors. Despite several differences, the distribution of HLA haplotypes in families of the patients and in donor registries is similar, and the probability of finding a compatible donor for patients with common HLA-haplotypes in Russian registries is quite high. Most of 7-locus haplotypes are associated with different alleles of the HLA-DP locus in the 9-locus haplotypes, due to presence of a recombination hot spot. The study revealed strong linkage disequilibrium between the HLA alleles DRB1*03:01 and DPB1*01:01P (D’ = 0.579), DRB1*07:01, and DPB1*17:01 (D’ = 0.808), DRB1*09:01 and DPB1*04:02P (D’ = 0.502). The information obtained about real 7- and 9-locus HLA-haplotypes in families may be used in clinical practice as a reference for analyzing the results of HLA-typing and predicting the expected HLA-haplotypes. It has been shown that, despite recombination hot spot between the HLA-DP locus and the rest of the HLA complex, there is strong linkage disequilibrium between some alleles of the DRB1 and DPB1 genes.

Glaucoma is a degenerative disease of the optic nerve, accompanied by the death of retinal ganglion cells (RGCs) and loss of vision. An important role in the pathogenesis of glaucoma is ascribed to activated microglia, which produce pro-inflammatory interleukins and initiate GCS apoptosis. It has been established that single nucleotide polymorphisms of interleukin genes modify the development of neuroinflammation, but their effect on the risk of developing glaucoma is not yet fully established. Our aim was to determine the pathogenetic role of gene polymorphisms in TNFα and IL1β in the development of primary open-angle glaucoma.

We have observed 56 patients of Russian nationality from the South of Russia with primary open-angle glaucoma (POAG), 28 patients with stage I, 16 with stage II, 12 with stage III POAG. The single nucleotide polymorphisms TNFα 308G>A (rs1800629) and IL1β -31 Т>С (rs1143627) were studied by restriction fragment analysis of PCR products. The level of pro-inflammatory cytokines (TNFα and IL1β) in the lacrimal fluid of patients with POAG was evaluated by enzyme-linked immunosorbent assay (Vector-Best test system). To perform optical coherence tomography by analysing the thickness of retinal nerve fiber layer (RNFL) with volume and area of the neuroretinal rim using Torson 3D OST 1000 apparatus.

Results: in patients with POAG, we have found more common incidence of TNFα 308A (OR = 5.21, p = 0.001), and IL1β-31 T alleles (OR = 1.99, p = 0.04). An increased risk of developing POAG was found in carriers of genotypes 308A/A (OR = 6.30, p = 0.049), 308G/A (OR = 3.60, p = 0.049) and -31T/T (OR = 2.67, p = 0.04). The highest levels of TNFα were determined in the 308A/A group (190 (153.0-220.0) pg/mL), IL1β were in the group (-31) T/T – 6.50 (4.10-7.00) pg/mL. A decreased thickness of the retinal nerve fibers was observed in the patients with TNFα G308A genotype (59.5; 40.0 to 78.0 µm, p = 0.03), and in TNFα A308A carriers (79.0; 65.0 to 80.0 µm, p = 0.001).

The TNFα 308 G/A (rs1800629), along with IL1β, -31Т/C (rs1143627) cytokine gene polymorphisms are associated with development of primary open-angle glaucoma. TNFα 308A, IL1β -31T alleles, as well as the 308G/A, 308A/A and -31T/T genotypes seem to be the risk factors for POAG in Russian population. High content of TNFα in the lacrimal fluid was found in the carriers of 308A/A genotype and -31T/T IL1β genotype. The lowest thickness of the retinal nerve fiber layer was observed in the carriers of tTNFα A308A and TNFα G308A genotypes.

Combined cardiological and ophthalmological pathology has a high prevalence in older age groups of the population and common pathogenetic mechanisms, among which, of course, is a violation of the cytokine profile. However, the cytokine profile of the blood was practically not analyzed in elderly patients with combined ischemic heart disease with glaucoma. The aim of the study was to study the cytokine profile in patients with combined cardio- and ophthalmopathology. The study was performed at the S.N. Fedorov National Medical Research Center “MNTK Eye Microsurgery”, in two groups: patients with combined coronary heart disease with glaucoma (n = 58 people), and patients with coronary heart disease (n = 49 people), who in both cases have the same age of 60-74 years. The diagnosis of glaucoma was carried out in accordance with the criteria of the “National Glaucoma Guidelines”. Electrocardiographic, echocardiographic, radiographic, and enzyme studies were performed to diagnose coronary heart disease. The determination of cytokines in blood plasma was carried out on the device “Becton Dickinson FACS Canto 2 (USA)” using a special set of CBA (BD Biosciences, USA). Among the patients of the compared groups of the same age, significant differences in most cytokines were revealed, namely, a predominant increase in patients with combined cardio- and ophthalmopathology relative to the group with coronary heart disease. The content of IL-5, IL-12, IFNγ, TNFα in the blood plasma of patients with coronary heart disease combined with glaucoma increased with a significant difference compared to patients with coronary heart disease. However, the highest increase among the cytokines under consideration is characteristic of IL-6 and IL-17, which amounted to 23.8±1.1 pg/mL and 20.2±1.7 pg/mL in patients with combined cardio- and ophthalmopathology versus 6.3±0.3 pg/mL and 7.9±0.5 pg/mL, respectively, in patients with coronary heart disease. At the same time, the level of IL-4 and IL-10 decreased significantly to 2.2±0.2 pg/mL and 6.4±0.4 pg/mL versus 4.8±0.3 pg/mL and 11.9±0.6 pg/mL. The use of logistic regression made it possible to determine the relative risk values of the studied blood cytokines and to develop uncorrected and adjusted models, according to which the closest association with the risk of developing combined coronary heart disease with glaucoma was established for IL-6 and IL-17, with the relative risk values in the uncorrected model of 2.87 and 2.71, respectively (p < 0.001). However, in the adjusted model, the association of IL-6 with combined coronary heart disease with glaucoma increased to 2.92 (CI 2.80-3.27, p = 0.004), and IL-17 decreased to 2.64 (CI 2.51-2.85, p = 0.003). There was also a significant association of IL-4, IL-5, IL-12, IFNγ and TNFα with combined coronary heart disease with glaucoma. The study demonstrated new associations of systemic cytokines with the risk of developing combined coronary heart disease associated with glaucoma.

Comorbid diseases which include arterial hypertension (AH) accompanied by overweight, and vibration disease (VD) are known to contribute to the mutual aggravation of occupational and cardiovascular pathology. Despite numerous studies indicating that some changes of immune system are observed when exposed to vibration, the contribution of cytokines to the comorbid course of vibration disease has not been sufficiently studied. The aim of the work is to evaluate the role of inflammatory mediators in development of hypertension and obesity in vibration disease.

The cytokine profile parameters were determined by ELISA immunoassay using “Vector-Best” test systems. Some unidirectional changes in serum cytokine concentrations were found in patients with VD complicated

by AH, and those with VD without AH, being characterized by increase in pro-inflammatory IL-1β, TNFα, IL-17, anti-inflammatory IL-4 and IFNγ, and a decrease in multifunctional IL-2 cytokinem, relative to the comparison group. The groups with versus without AH differed in compensatory increase of anti-inflammatory IL-10 in the latter group. At the same time, all patients with combined VD and AH had excessive body weight, with grade 1 obesity in half of the cases, thus being significantly different from those with VD without AH. It is shown that IL-1β, IL-2, IL-4, IL-8, TNFα play an apparent role in the development of obesity. The greatest contribution is observed for IL-1β, as evidenced by the quantitative relationship of body mass index with its concentrations as assessed by logistic regression analysis. The limitation of this study may be a small sample size.

It was found that the unidirectional changes in cytokines when compared to the comparison group were revealed in patients with VD complicated by AH, and in patients with VD without AH. In the subjects with VD and AH, these changes were more pronounced, which may suggest the hypertension is an aggravating factor. The established relationship between individual cytokines (IL-1β, IL-2, IL-4, IL-8, TNFα) and BMI indicates their participation in development of obesity. All these findings may provide a basis for development of preventive and therapeutic measures that reduce the risk of hypertension and obesity in the people working under permanent exposure of vibration.

Resistance to tyrosine kinase inhibitors (TKIs) is currently an important clinical problem in the management of patients with chronic myeloid leukemia (CML). Recent studies suggested that aberrant cytokine secretion may be among the BCR/ABL-independent mechanisms of resistance, thus contributing to the persistence of leukemic stem cells in spite of continuous targeted therapy. The aim of the study was to evaluate concentration of cytokines in the serum of patients with CML depending on the efficiency of therapy.

Quantitative determination of the cytokines (TNFα, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-17, IL-18, IFNα and VEGF) in blood serum of patients with chronic-phase CML (n = 84) and healthy subjects (n = 30) was performed using enzyme immunoassay (ELISA). The patients with CML were divided into 3 groups depending on the duration of therapy: group I, newly diagnosed patients (n = 10); group II, patients receiving therapy for < 12 months (n = 10); group III included patients receiving therapy for more than 12 months (n = 64).

The results of our study showed that cytokine concentration among CML patients significantly differed, depending on the duration of therapy. Significantly higher concentration of IL-17, IL-6, IL-1β, IL-10, IL-18, IL-2 and TNFα was found in group I compared with control group. Group II patients also demonstrated significantly higher concentrations of TNFα, IL-6, IL-10, IL-18 and IFNα by comparison with control group, as well as higher concentration of IFNα compared with in groups I and III. In group III, concentrations of IL-17, IL-1β, TNFα, IL-6, IL-10, IL-18 were significantly higher than in control group. When compared with group I, it was found that concentrations of IL-1β, IL-2 and IL-18 were significantly lower. A direct correlation was found between expression levels of chimeric BCR/ABL gene, (a marker of CML malignancy), and concentrations of IL-1β and IL-17. ROC-analysis demonstrated high-quality models which showed an association between achievement of major molecular response (MMR) and low serum concentrations of IL-1β, IL-6 and IL-17.

Hence, the results of our study have shown that determination of IL-1β, IL-6 and IL-17 concentrations may be a prognostic marker for assessing the efficiency of therapy and probability of achieving MMR in CML.

Cytokines are the key mediators which control and regulate immune and inflammatory responses via complex networks and serve as biomarkers of many diseases. Quantitative determination of cytokines is helpful in assessing immune status and adjusting therapy for various inflammatory diseases, such as sepsis and pneumonia. Since community-acquired pneumonia remains a common cause of childhood morbidity and mortality. At the present stage, prognosis of the disease severity in children is an urgent problem. The aim of our study was to identify associations between cytokine levels in healthy children and in children with community-acquired pneumonia (CAP), depending on the age of patients and severity of the pathological process. The work was carried out at the Department of Microbiology, Virology and Immunology, Department of Propaedeutics of Childhood Diseases and Pediatrics, and at the Research Institute of Immunology at the South Ural State Medical University. The study included 117 children aged 1 to 18 years with radiologically confirmed diagnosis of CAP, either severe or mild degree. The comparison group was composed from 28 healthy children who did not have community-acquired pneumonia, as well as other signs of acute respiratory viral infection at the time of examination, being observed for any chronic disorders. The levels of IL-1β, IFNγ, IL-6, IL-4, IL-10, IL-2, TNFα, IFNλ2 (IL-28A), IFNλ3 (IL-28B), IL-8, MCP-1, IL-17AF, GM-CSF were determined in blood serum by means of ELISA test systems (a “sandwich” technique with peroxidase as an indicator enzyme). For statistical analysis, a multidimensional method was used, i.e., non-linear analysis of the principal components using the CATPCA algorithm. Among the children with CAP, our study revealed a consistent increase of IL-1ß, IL-4, IL-10, IL-2, TNFα, IFNλ2 (IL-28A), IFNλ3 (IL-28B), IL-8, MCP-1, IL-17AF, GM-CSF in blood serum. The highest correlation with severity was shown for IFNλ2 (IL-28A), IFNλ3 (IL-28B) and MCP-1, which may be considered additional biomarkers of the CAP severity. There was also a significant variability of the cytokine profile in healthy children and its significant skewing in pneumonia, especially in severe cases.

An important area of research concerns monitoring of immune response features in patients with SARS-CoV-2 infection as well as their analysis, as compared with characteristics of vaccine-mediated protection, in order to specify the determinants of cellular immune response. The aim of our work was to compare the state of cellular immune response in patients who underwent COVID-19, and in persons vaccinated with a peptide vaccine preparation. The study involved volunteers who suffered with COVID-19 of varying severity (n = 30), as well as persons who completed the full course of vaccination with the peptide vaccine (n = 27). For comparison, we took blood specimens from the volunteers before vaccination. Immunophenotyping of leukocytes was performed by the Lyse/No-Wash procedure (BD Bioscience, USA), and Cyto-Stat monoclonal antibodies (CD45-FITC, CD4-PE, CD8-ECD, CD3-PC5), CD45RA-PC7, CD45RO-PE (Beckman Coulter, USA), and analyzed with a DakoCytomation flow cytometer (Denmark). Determination of intracellular IFNγ (CD4⁺IFNγ⁺) was performed with the standard technique. Cytokine production was determined using reagent kits for detection of IFNγ, TNFα, IL-4, IL-8, IL-10 (Vector-Best JSC, Russia) with automatic enzyme immunoassay analyzer LAZURIT (Dynex Technologies, USA). As based on the results obtained, we have shown that cellular immunity was developed after vaccination and infection with COVID-19. However, the most pronounced immune response was recorded in the COVID-19 reconvalescents, i.e., more than 60% of these patients showed an increased number of CD4⁺T-memory helper cells (8.7 (0.5-12.1) % versus 0.3 (0.1-0.5) % in the comparison group, p < 0.05) as well as proportion of CD4⁺IFNγ⁺T lymphocytes (4.2 (1.8-4.3) % versus 0.4 (0-0.8) % in the comparison group, p < 0.05). Moreover, we revealed an increased functional reserve of cells in terms of TNFα, IL-8, IL-10 production. One month after vaccination of volunteers with the peptide-based preparation, the total pool of memory T lymphocytes was apparently dominated by CD8⁺T memory cells (CD45⁺CD8⁺CD45RA^-CD45RO⁺). A significant increase was found in the average levels of CD4⁺IFNγ⁺ activated cells (8.2-fold), as well as in values of ConA-induced IL-4 production (3.3 (1.1-4.5) pg/mL, and 2.8 (1.7-3.9) pg/mL, respectively versus 1.3 (0.1-2.4) pg/mL in the control group, p < 0.05). The data obtained are in accordance with information available in the literature concerning development of cellular immune responses to SARS-CoV-2, which results from a past illness, or measures for the specific prevention of COVID-19. Further search for cellular correlates of protection against a new coronavirus infection will allow us to revise the current vaccination strategy and develop an optimal approach to COVID-19 prevention.

Cellular immunity plays an important role in the control of SARS-CoV-2. Lymphopenia and a decrease in the functional activity of cells may be among the main reasons for deterioration of clinical outcomes of the disease. Usage of the bacterial therapeutic vaccine Immunovac-VP-4 during the inflammation phase may be promising for immunomodulation of the cellular immunity. The aim of our study was to evaluate the dynamics of lymphocyte subpopulations in hospitalized patients with COVID-19 upon combining the basic therapy with immunotropic drug based on the antigens from opportunistic pathogens. The study included 45 patients (18-70 years old) admitted with a confirmed diagnosis of moderate/severe infection caused by the COVID-19 virus. In addition to basic therapy, 33 persons of this group received Immunovac-VP-4 by a combined nasal-oral method. Subpopulation activity of peripheral blood lymphocytes in patients over time (at baseline, on the 14th and 30th day after hospitalization) was studied by flow cytometry by means of FC-500 Cytomics (Beckman Coulter, USA) using monoclonal antibodies (mAb) (Immunotech, France). In the group receiving only standard therapy, an increased number of T lymphocytes was detected on day 14 (79.9 (75.5-81.6), p = 0.00252), on day 30 from the start of treatment (78.4 (74.25-79. 2), p = 0.03662), and a decrease in B lymphocytes on day 14 (10.6 (7.78-11.63), p = 0.03236), on day 30 (7.85 (6.25-11.1), p = 0.01352) relative to baseline parameters upon admission. We revealed more pronounced changes in the parameters of cellular immunity relative to the initial parameters, i.e., an increased proportion of T lymphocytes on the 14^th day (80.1 (73.8-84.2), p = 0.00018), and 30^th day from starting the treatment (80.2 (76-81.9)), T helpers at 14 days after treatment (50.2 (43-57), p = 0.00694), cytotoxic T cells by 30^th day of therapy (26.35 (24-29.4), p = 0.0114), decrease in B lymphocytes on day 14 (13.1 (8.2-16.9), p = 0 00158), on the 30^th day from the start of treatment (8.2 (7.6-9.7), p <0.00001), and a transient decrease in NK cells on the 14^th day (3.7 (2,1-6.3), p = 0.00308), with their recovery on the 30^th day of observation to 8.6 (6-12.5) in the Immunovac-VP-4 group. Modulation of cellular immunity may be important for the virus clearance.

Chronic endometritis with impaired receptivity of uterine cavity epithelium (the “thin” endometrium) is considered the most common cause of recurrent implantation failures during in vitro fertilization (IVF). Chronic inflammation, accompanied by autoimmune mucosal reactions, may lead to a perverse immune response of lymphocytic cells with altered cascade of cytokine reactions, thus preventing efficiency of decontamination therapy, followed by potential inability to perform reproductive function in the patients. The article concerns our experience of complex preimplantation preparation of patients with recurrent implantation failures caused by chronic endometritis during a course of intrauterine administration of cavitated solution of recombinant interleukin 2 (rIL-2). Our aim was to assess some changes in local immune status in the patients with chronic endometritis and repeated implantation failures during complex preparation for the IVF procedure, using a cavitated solution of rIL-2.

A study of the local immune changes of uterine cavity in the patients with recurrent implantation failures suffering with chronic endometritis was carried out under complex preparation for IVF with thawed embryos (n = 82). A control group included the infertile women with normal endometrial thickness (> 7 mm at M-echo), with excluded diagnosis of chronic endometritis (n = 30). Complex therapy included decontamination and contamination; hormone replacement therapy; intrauterine ultrasonic cavitation of rIL-2 solution. Dynamic assessment of the local immunity was tested in the endometrial biopsies. We performed analysis of the main subpopulations of immune lymphoid cells, morphological composition, phagocytic responses and cytokine status.

he state of immunocompetent cells and cytokine profile of endometrial samples in the patients with chronic endometritis indicates a possible Th2 shift of the local immune response, with changing ratio of cytokines leading to impaired molecular, subcellular and cellular structures, which, along with fluctuating activity of other components of uterine cavity homeostasis, may determine a recurrent course of the disease with impaired endometrial morphology. Preimplantation preparation with intrauterine irrigation with a cavitated solution of rIL-2 in patients with recurrent implantation failures accomplished by chronic endometritis helps to improve the indices of local immunity, being more effective (an average of twofold), compared with the group of conventional preparation for IVF with thawed embryos. The treatment may promote the proliferative processes in epithelium of the uterine cavity thus contributing to increased potential of endometrium for implantation and occurence of clinical pregnancy.

Long-term persistence of inflammation is the main factor of pathogenesis in chronic polypous rhinosinusitis (CRSwNP). Prospectives of current clinical otorhinolaryngology include modification of drugs containing topical glucocorticosteroids in order to increase local bioavailability, reduce the concentration of glucocorticosteroids, and alleviate the local inflammatory response. The aim of our work was to evaluate the effect of urea, magnesium sulfate, and mannitol on the concentrations of mometasone furoate and IL-5 in the polypous tissue of patients with CRSwNP. The study included 146 patients with CRSwNP aged 18 to 62 years. Biopsies of polypous tissue were taken during polypotomy. In the fluid samples obtained by microdialysis of polypous tissues, mometasone concentrations were determined by high-performance liquid chromatography. Concentrations of IL-3, IL-4, IL-5 and IL-10 cytokines in blood serum, and IL-5 in microdialysates of polypous tissue were measured by ELISA technique. The average age of patients with CRSwNP was 42.9±7.2 years, with prevalence of male patients (n = 93, 63.7%) over females (n = 53, 36.3%). Severity parameters of the disease by the SNOT-22 scale were significantly higher in CRSwNP patients than in control group (p ≤ 0.05). The results of biochemical analysis of microdialysates from polypous tissue showed increased content of Na⁺ ions in the resistant clinical cases of CRSwNP compared with the sensitive and dependent clinical forms (p < 0.05). Curves with the level of mometasone furoate were obtained in all the studied samples. It was found that the addition of magnesium sulfate solution to mometasone increases the Cmax of mometasone to 154 ng/mL in biological fluids after microdialysis. When urea solution is added to mometasone, Cmax increases to 198 ng/mL. In the native nasal polypous tissue dialysate, IL-5 levels were 89±2.01 pg/mL. Two hours after the addition of mometasone furoate, IL-5 concentrations decreased to 61±3.5 pg/mL, upon addition of urea, it changed to to 69±2.98 pg/mL (p ≤ 0.01, compared with the control without the addition of drugs). Analysis of serum cytokine levels showed that IL-5 concentrations in the patients with CRSwNP significantly exceeded (> 3-fold) those in the control group (p ≤ 0.05). There was a trend for increase of IL-3 and IL-4 (2-fold) and to decreased IL-10 levels in blood sera (> 1.5-fold) in patients with CRSwNP compared with controls. These results may be helpful for development of novel clinical approaches, in order to increase bioavailability of topically administered glucocorticosteroids, thus allowing to promote the effectiveness of basic therapy of CRSwNP and to develop therapy for the prevention of resistant forms of polyposis.

Studies on pathogenesis of fetal membrane insufficiency in preterm pregnancy should expand the opportunities of predicting prenatal rupture of amniotic fluid and improve the strategy of anticipant gestation management in cases of premature rupture of the fetal membranes. The clinical significance of studying this obstetric problem is unquestionable due to the high risk of complications and perinatal losses caused by preterm birth. This research is devoted to studying the changes in cytokine profile of amniotic fluid during prenatal rupture of amniotic fluid at 22 to 34 weeks of gestation. The levels of pro-inflammatory (TNFα, IL-6, IL-1β, IL-2, IL-6, IL-8) and anti-inflammatory (IL-4, IL-10) cytokines in amniotic fluid were determined in 30 patients, whose premature pregnancy was complicated by early rupture of the membranes. For reference, the level of these cytokines in the amniotic fluid was studied in 25 pregnant women with a physiological course of pregnancy at full-term gestation. The ELISA technique was used with a test system produced by JSC Vector-Best. The study was carried out on the basis of the Perinatal Center of the Saratov Region. We have found that premature rupture of the membranes is preceded by increased levels of pro-inflammatory cytokines in amniotic fluid. The latter result suggests a significant alteration in feto-placental complex, which may be the starting point both for damage to the fetal membranes, as well as for development of labor activity.

Low-energy fractures in rheumatoid arthritis (RA) are more common in patients with high activity and long duration of disease, and with high titers of anti-citrullinated antibodies (ACPA). Increased expression of angiopoietin-like protein type 4 (ANGPTL4) in bone tissue has also been noted in inflammatory arthritis. The purpose of the present study was to analyze the effect of ACPA and ANGPTL4 on systemic bone mineral density in RA patients. Antibodies to ACPA and ANGPTL4 content were detected in blood serum of 96 RA patients (women, 91.7%) by enzyme immunoassay. Mineral density of the lumbar vertebrae (BMD_L1-L4), hip neck, and entire femur (BMD_total) was measured by dual-energy X-ray absorptiometry (DXA). In study group, the ACPA and ANGPTL4 tests were positive in 61.5% and 41.7% of patients, respectively. Negative correlations were shown between ACPA and BMD_total, and of ANGPTL4 with BMD_L1-L4 (p < 0.05). Separation of the patients into groups with low (n = 34) and high (n = 62) DAS28 activity demonstrated a significant increase in ACPA with increasing RA activity (p = 0.042). ACPA and ANGPTL4 scores were also significantly higher in the group of 45 RA patients with osteoporosis (OP) compared to the RA group without OP (n = 51) showing significant difference at p = 0.002 and p = 0.028, respectively. Patients’ age, body mass index (BMI), duration and activity of the disease had no significant effect on ACPA in the general group of RA patients. However, the correlation between ACPA and DAS28 proved to be significant in the group of RA patients with OP (b = 0.31, p = 0.039). Among all presented variables, the disease duration was the only significant factor for ANGPTL4 in the total group of RA patients (b = 0.31, p = 0.039). In the regression model, BMD_total showed similar correlations with patients’ age (b = -0.28), BMI (b = 0.25), and ACPA level (b = -0.26). A search for association between BMD_L1-L4 and various RA characteristics demonstrated a strong correlation with ANGPTL4 only (b = -0.74; R² = 0.57). The revealed correlation between ANGPTL4 and decreased BMD specifically in the spongy layer of bone allows us to identify the RA patients with high ANGPTL4 levels as a risk group specifically for spinal fractures thus considering ANGPTL4 as a potential target for treatment of osteoporotic disorders.

Diseases caused by Streptococcus pyogenes are one of the significant problems of practical health care, due to the ability of this pathogen to cause morbidity in all age groups of the population with the development of different complications. The streptococcal infection is accompanied, along with infectious and inflammatory symptoms, by a pronounced immune response to Streptococcus pyogenes, which is mediated by the functional activity of monocytes and dendritic cells responsible for the production of pro- and anti-inflammatory cytokines that regulate the interaction, proliferation and functional activity of all participants in the cellular and humoral links of immunity. The purpose is to study the effect of a pyrimidine derivative on the level of pro- (IL-1β, IL-6, IL-8) and anti-inflammatory interleukins (IL-4, IL-10) in conditions of experimental streptococcal wound infection. The study of the effect of the pyrimidine derivative 3-[2-[(4,6-dimethylpyrimidin-2-yl) amino]-2-oxoethyl]quinazolin-4(3H)-one was carried out in vivo on the model streptococcal wound infection. The experiments were carried out on CBA mice 3 months old (20-22 g). Animals were divided into groups: control I–healthy animals, which were injected with water for injection; control II–infected untreated animals; experience I – animals that were treated intraperitoneally with a pyrimidine derivative at a dose of 36 mg/kg for 7 days, starting from the first day of infection; experiment II – animals that were treated intraperitoneally with the reference drug cefepime at an average therapeutic dose of 50 mg/kg in the same regimen as the test compound. The levels of pro- and anti-inflammatory interleukins in blood serum were determined by enzyme immunoassay. The study found that the pyrimidine derivative 3-[2-[(4,6-dimethylpyrimidin-2-yl)amino]-2-oxoethyl]quinazolin-4(3H)-one under conditions of streptococcal wound infection has an immunoregulatory effect, manifested in a decrease in pro-inflammatory and an increase in anti-inflammatory interleukins.

Cervical cancer is the most common malignant tumor of the female genital organs. In general, the prognosis in patients with advanced cervical cancer is unfavorable. The option of choice for stage IV of the disease and relapses is systemic platinum-containing chemotherapy. Its effectiveness is about 20-26%, life expectancy is 12 to 13 months. Undoubtedly, the search and development of new methods of treating this disease is an extremely urgent task. Immuno-oncology has emerged as a potential new strategy to improve the treatment outcomes of patients with malignant neoplasms. Much attention in research is focused on the opportunity of using interleukin-2, tumor necrosis factor (TNF) and interferon-gamma (IFNγ) for tumor immunotherapy, since these cytokines play a special role in antitumor protection. Due to the active search for new hybrid molecules based on TNFα, some domestically developed recombinant antitumor drugs are implicated into modern practice of clinical oncologists, in particular, “Refnot” (TNFα-thymosin-α1) and “Ingaron” (IFNγ preparation). We analyzed the result of combined treatment (standard polychemotherapy at the 1st line augmented with cytokinotherapy including Refnot + Ingaron therapy) in one patient with recurrent cervical cancer. According to the control examination, upon completion of the polychemotherapy course, a complete tumor response was registered according to the Recist 1.1 criteria. The patient continued to receive cytokines as supporting therapy. Currently, according to control quarterly examinations, a complete regression of recurrent tumor persists from the end of polychemotherapy to 28 months of observation. One should note that when monitoring the state of the immune system during therapy with Refnot and Ingaron, we noted an increase in absolute and relative numbers of T cells to normal levels along with higher cytotoxic and antitumor potential of NK cells without increasing their number. The patient well tolerates the therapy, improved quality of life is documented, and there are no clinically significant side effects. Hence, the therapy with “Refnot” (TNFα-thymosin-α1) and “Ingaron” (IFNγ) in this clinical case proved to be a safe method of maintenance therapy with a positive therapeutic effect thus allowing effective control of recurrent cervical cancer for more than 2 years, as well as significantly improve quality of life of the patient. This type of therapy may be recommended for usage in clinical oncology.