Sepsis is a heterogeneous and life-threatening condition caused by a dysregulated immune response to infection. The most severe form of sepsis is septic shock, characterized by arterial hypotension, impaired tissue perfusion, and hypoxia. Despite new findings in antimicrobial and intensive care therapy, the incidence and mortality rates of sepsis remain high, which underscores the relevance of further studies on its pathogenesis. In recent years, the research has shifted from clinical signs to analysis of immunological and molecular mechanisms, which allowed to identify specific phenotypes and endotypes of the disease. Sepsis phenotypes are based on clinical manifestations and biomarkers, whereas its endotypes are defined by molecular mechanisms, including immune gene expression patterns. This article reviews key aspects of the innate and adaptive immune responses in sepsis, including activation of proinflammatory cytokines, development of coagulopathies, deterioration of endothelial integrity and microvascular regulation. Moreover, the potential mechanisms, e.g., hyperinflammation, simultaneous development of immunosuppression, and functional exhaustion of immunocompetent cells are highlighted. Thus, immunological biomarkers are considered a promising tool for patient‘s stratification, prognosis prediction and personalized therapy. Current immunodiagnostic methods are also discussed in this article, including quantitative analysis of cytokine levels and assessment of innate immune dysfunction markers. Hence, current understanding of sepsis as an immunologically heterogeneous syndrome enables researchers to expand existing concepts of its pathogenesis. In contrast to classical concept based on a shift from inflammation to immune suppression, the novel results suggest a simultaneous presence of both processes in the same patient, making it necessary to reconsider the existing diagnostic and therapeutic approaches.

Functioning of soluble receptors, mainly soluble cytokine receptors, has been of interest to researchers since the last decade of the 20th century. Allergists’ interest in the issues of soluble receptors is relevant to our studies in the pathogenesis of bronchial asthma (BA), specifically, development of ectopic chemosensory signaling systems, including ectopic bitter taste receptors Tas2R, and signaling pathways of the Receptor for Advanced Glycation End products (RAGE). The expression of soluble receptors in BA was investigated in both studies. The article discusses the mechanisms of soluble receptor formation: 1) proteolytic cleavage of existing membrane receptors; 2) synthesis and release of soluble receptors lacking a transmembrane domain via alternative mRNA splicing; 3) release of membrane receptors in exosomes. Possible functional effects of soluble receptors are considered, i.e., neutralization of corresponding ligands (decoy receptors), ligand transport, ligand stabilization, and binding of membrane proteins. The results of studies of soluble receptors in health and disease are sometimes contradictory and not fully understood. Soluble receptors may be considered prospective therapeutic agents. Our previous data concerning probable functions of a number of soluble Tas2R38, Tas2R31, Tas2R5 receptors are considered when studying the patients with asthma in view of earlier knowledge of the soluble receptors functioning. The most important characteristics of the soluble receptors studied in BA are: 1) lower plasma levels of all discussed Tas2Rs in allergic asthma compared to nonallergic forms; 2) an inverse correlation between plasma Tas2R levels and bronchial resistance; 3) correlation of Tas2Rs with monocyte-macrophages, granulocytes and bronchial ciliated cells. The regulatory role of soluble receptors involved in asthma pathogenesis, mainly cytokines, as well as sTas2R and sRAGE, can be viewed through the corresponding membrane receptor functions. In this case, soluble forms of receptors play the role of decoy proteins that block functions mediated by membrane receptors. In this approach, the discussed soluble cytokine receptors and sRAGE have a positive regulatory effect in BA, whereas sTas2R exert negative action. The balance of soluble and membrane-bound receptor functions in cells involved in asthma pathogenesis is crucial for developing the novel therapies for asthma by targeting the Tas2R and RAGE receptors.

Hypoxia can be both a cause and a consequence of pathogenic events in the infectious, autoinflammatory, and autoimmune processes. Given that human and animal populations are genetically heterogeneous in terms of whole-body resistance to oxygen deficiency, the modern approach to predicting and treating diseases associated with impaired immune regulation requires taking into account the role of hypoxia among pathogenetic mechanisms of inflammation. The purpose of the study was to evaluate the features of neutrophil effector functions under normal conditions and in presence of an induced inflammatory process in animals with genetically predetermined high and low resistance to hypoxia. For our studies, we used 8-monthold male rats of two breedings with different genetic tolerance to hypoxia (highly resistant HR/SmY line; and low-resistant LR/SmY line), weighing 400-450 g. Rats from the experimental groups of the HR/SmY and LR/SmY lines were induced to develop immune responses in a model of rheumatoid arthritis (RA). After 35 days, the blood samples from the animals were incubated with Indian ink suspension (1:10). The smears fixed in formalin vapor were stained with 0.5% methylene blue solution and analyzed with light microscopy (400×). Phagocytic index (PHI), phagocytic number (PN), and the number of suicidal netosis with partial and complete chromatin decondensation were calculated. The significance of differences in the groups was assessed by the Mann–Whitney test and the Student’s t-test. PHI and PN of neutrophils did not significantly differ between control groups of LR/SmY and HR/SmY animals (52.5%/49%; 1.68/1.80, respectively). In experimental groups, the PHI of neutrophils of LR/SmY line (66%, p ≤ 0.05) exceeded that of the HR/SmY line (56%). With systemic inflammation model, the PHI of neutrophils in the group of low-tolerance rats increased by 1.26 (p ≤ 0.05) times, and the number of neutrophils capturing 2-8 particles increased by 1.3 (p  0.01) times in healthy rats. In experimental HR/SmY group, 1.81% of more active neutrophils absorbing 9 to 12 particles (p ≤ 0.01) were recorded, with PHI was 1.2 times lower than that of the LR/SmY line. In healthy HR/SmY rats, higher ratio of neutrophils (1.4-fold increase) was involved in the process of suicidal netosis than in similar group of the LR/SmY line (p ≤ 0.05). Systemic inflammation in the LR/SmY group caused a twofold increase in NETs (19.67%), which was 1.7 times higher than in the experimental HR/SmY group. The organisms with genetically determined low resistance to hypoxia, when accompanied by inflammatory process, show higher burden on cell immunity in the course of phlogogen elimination as compared to highly resistant ones, which may predispose them for more severe pathogenic forms of inflammatory and autoimmune diseases.

Current experimental data indicate the leading role of cytokines in the formation of manifestations of bronchial asthma (BA), thus supporting the so-called anti-cytokine approach to the therapy of this disease. The aim of this study was to evaluate possible association of rs10455025 TSLP gene variant and rs11811856 polymorphism of TNFSF4 gene with the risk of developing BA among children from Kursk population. The study included 999 unrelated individuals, including 526 patients with BA and 473 control persons. These functional gene polymorphisms (SNPs) were genotyped using the MassArray-4 genomic mass spectrometer. We have found that the alleles rs11811856G TNFSF4 and rs10455025C TSLP, as well as the genotypes rs11811856G/G and rs10455025-C/C are associated with an increased risk of developing BA in children. The association of rs11811856 TNFSF4 with an increased risk of developing BA was revealed in both boys and girls. Meawhile, the rs10455025 polymorphism of TSLP gene did not show any associations in the stratified analysis by gender. Upon analysis of gene-environment interactions in general group, a relationship was established with the risk of asthma depending on exposure to tobacco smoke and the place of residence of children. We have found a relationship between the SNP status and the risk of concomitant allergopathology in children with BA, and the influence of the studied polymorphisms on laboratory indices of rhinocytogram and spirometry. The results of this study have shown that the studied SNPs (rs10455025 TSLP and rs11811856 TNFSF4) in children with BA are associated with increased risk of developing food sensitization to banana, rice, duck, and a number of pollen (foxtail, common horsetail, bluegrass, ryegrass, fireweed, ash, barley), and epidermal (human hair) allergens. It has also been found that the studied SNPs reduce the risk of sensitization to house dust, cat hair, rabbit hair, and quinoa pollen. The present study showed that the polymorphic gene variants (rs10455025 TSLP and rs11811856 TNFSF4) are significantly associated with the risk and clinical features of developing BA.

In 2020, the COVID-19 pandemic has been declared. Since then, it was revealed that the severity of the disease pathology may depend not only on the viral strain, but also on the functioning of immunity system. The aim of our study was to evaluate some characteristics of expression and genetic factors of innate immunity in patients who have had COVID-19. We have studied scrapings from mucous membranes and venous blood samples taken from 148 patients. Identification of the studied markers was carried out using reverse transcription and RT-PCR methods. Statistical analysis of results was carried out using Mann–Whitney test, Fisher’s exact test, χ² test, odds ratio and 95% confidence interval. Our study has demonstrated a prognostic role of polymorphic markers, i.e., haplotypes of TLR9 (rs352140 and rs5743836) and TLR4 (rs11536889 and rs4986791) gene variants related to the risk of severe SARS-CoV-2 infection. When studying long-term effects of COVID-19, we have revealed that an imbalance in the expression of receptor and effector molecules at the mucosal level of immune response remains in patients who had a history of this disease. There was a decreased expression of both receptor molecules (TLR3, TLR7) and antiviral immune response factors (IL-28) in the oropharyngeal mucosa along with general increase of these markers in epithelial cells of nasopharyngeal mucosa. Expression levels and genetic factors of innate immunity associated with severe SARS-CoV-2 infection and, consequently, persistent changes in immune system long after recovery, have been revealed, thus expanding our knowledge of molecular genetics mechanisms associated with prolonged course of COVID-19. The results obtained during the study may help to assess the risks of developing severe infection caused by SARS-CoV-2 and evolving complications in hospitalized patients.

Rhinosinusitis is one of the most common diseases in otolaryngology which manifests itself as nasal congestion, headache, nasal discharge and other symptoms. Sometimes rhinosinusitis accompanies seasonal acute respiratory viral infections, being a manifestation of seasonal and year-round allergies, and develops with dysfunction of the immune response. Chronic rhinosinusitis (CRS) is diagnosed if the symptoms of rhinosinusitis last for >12 weeks. Comorbid conditions (curvature of the nasal septum, bronchial asthma, etc.) increase the manifestations of CRS. The purpose of our work was to study the features of cytokine regulation in CRS with comorbid diseases. 84 persons with rhinosinusitis and 100 healthy controls were involved. The study included patients with CRS (n = 29), CRS and deviated septum (n = 24), CRS with bronchial asthma (n = 31) and a control group (n = 100) consisting of practically healthy individuals. The case–control study was performed in Laboratory of Clinical Pathophysiology at the Research Institute of the Railway Ministry. The studied materials were venous blood and nasal discharge. The content of TNFα, IFNγ, IL-2, IL-6, IL-4, IL-10 was determined by ELISA technique using JSC "Vector-Best" reagents (Novosibirsk, Russia) and a Muitiskan FC spectrophotometer. Statistical processing was performed using the Statistica 10.0 program. To describe samples with a distribution different from normal, we used the median (Me) and interquartile range (IQR), defined as the difference between the 25^th (Q_0.25) and 75^th (Q_0.75) percentiles. This approach allows us to present the central tendency and measure of data dispersion, minimizing the impact of outliers (Wilcox, 2012). All patients with CRS and comorbid conditions systemically and locally show unidirectional changes manifesting as an increase in proand anti-inflammatory cytokines, thus suggesting the development of immune responses by Th1/Th2 mechanisms. The presence of comorbid conditions (curvature of the nasal septum, bronchial asthma) enhances the immune response. In comorbid conditions, the content of cytokines is statistically significantly increased relative to both control group and patients with CRS. Maximum concentrations of cytokines are detected when CRS was combined with bronchial asthma.

Chronic obstructive pulmonary disease (COPD) is one of the most common diseases of the bronchopulmonary system. A search of novel inflammatory markers for early diagnosis, optimization and monitoring of therapy is a promising direction of modern studies. Our objective was to evaluate the levels of YKL-40 and NGAL in serum and induced sputum and to determine their significance as inflammation markers in patients with COPD. The study included 50 patients with COPD, 60 patients with asthma. The control group consisted of 30 ageand sex-matched volunteers without allergic and broncho-obstructive diseases. Clinical and anamnestic data, indices of external respiratory function were evaluated. Induced sputum was collected with subsequent assessment of cellular composition. Immunophenotyping of lymphocytes by flow cytometry was performed. The concentration of YKL-40 and NGAL in serum was determined using an enzyme immunoassay test system (R&D Systems). The obtained data were processed using STATISTICA and SPSS Statistics software systems. A significant increase in the absolute number of T lymphocytes in patients of the studied groups compared to the control group was found. The highest number of T cytotoxic lymphocytes and NK-cells was registered in the COPD group. Cell population analysis in the induced sputum from COPD patients revealed the predominance of neutrophilic inflammation pattern. The maximum YKL-40 concentration in blood serum was registered in COPD patients. The levels of NGAL in serum of COPD patients were not significantly different from those in asthma group. NGAL level in induced sputum was significantly higher in COPD group. Correlation analysis confirmed a correlation between YKL-40 and NGAL levels in serum and neutrophilic inflammation indices, smoking index, number of hospitalizations due to COPD exacerbation during a calendar year. YKL-40 and NGAL levels were found to be significantly increased in patients with common COPD exacerbations compared to the group with infrequent exacerbations. YKL-40 and NGAL are promising markers of neutrophilic airway inflammation in COPD patients. The obtained data allow us to consider YKL-40 and NGAL as markers of non-T2-endotype COPD with neutrophilic inflammation pattern and high risk of exacerbations.

The purpose of our study was to evaluate the enzyme profile of purinergic system and lymphocyte subsets in patients with newly diagnosed infiltrative pulmonary tuberculosis (IPT), depending on the drug resistance of Mycobacterium tuberculosis (Mtb) to anti-tuberculosis drugs. In 109 patients with drug-sensitive Mtb (significant or less pronounced improvement after intensive phase of chemotherapy), or in drug-resistant cases, the activity of adenosine deaminase (eADA-1, 2), concentration of ecto-5’-nucleotidase (eNT5E), CD26 (DPPIV), and the composition of lymphocyte subsets were evaluated before treatment. The IPT patients with drug-sensitive Mtb strains who achieved a “less pronounced improvement” exhibited higher concentrations and activity of ectoenzymes responsible for production of extracellular adenosine (eNT5E) and its transformation (eADA-1 and eADA-2). The proportion of cytotoxic T cells was also higher compared with patients who achieved significant improvement. Patients isolating drug-resistant Mtb strains who achieved a “less pronounced improvement” had lower absolute counts of T lymphocytes and helper T cells with an increased proportion of cytotoxic T cells and elevated eADA-2 activity compared with individuals who achieved significant improvement. Thus, prior to initiation of tuberculosis chemotherapy, the activity of purine metabolism enzymes and the subpopulation profile of lymphocytes were not associated with the characteristics of Mtb drug resistance. A relationship between the parameters of purinergic regulation enzymes and numbers/ ratio of lymphocytes was revealed in patients who achieved significant improvement. Such relationships were not revealed in the group with less pronounced improvement, regardless of the drug resistance of Mtb. These findings suggest an imbalance of inflammatory factors and immune response to Mtb in the patients who showed worse clinical outcomes after intensive chemotherapy. Taking into consideration each component of protective reactions is required for administration of adequate chemotherapy, pathogenetic treatment, and immunocorrective treatment in order to prevent progression of the disease.

Despite a wide availability of specific prophylaxis against viral hepatitis B (HBV), the issues of HBV morbidity among healthcare workers remain quite sufficient. A latent form of hepatitis (LHB) may play a role in maintaining the epidemic spread of HBV. The aim of the study was to identify the dependence of the incidence of anti-HBc antibodies among employees of medical institutions by their age, gender and professional categories. Materials and methods: Blood serum samples were tested for anti-HBc in 1,643 medical staff members (doctors, nurses, paramedics, other personnel). Blood sera were examined for the presence of anti-HBc by ELISA using the HepaBest anti-HBc-IgG enzyme-linked immunosorbent assay system manufactured by Vector-Best-Europe JSC. The results were statistically analyzed using Microsoft Office Excel 2010 and Prism9 (GraphPad, USA). For comparison groups of medical workers according to the frequency of occurrence of Anti-HBc, the chi-square criterion was used. Results. Anti-HBs were detected in 601 persons, thus reaching 36.6% of the total study sample. In 1042 subjects (63.4%), anti-HBs were not revealed. Almost half of paramedical personnel from the study sample were IgG seropositive for the HBV nuclear antigen (50.6%). There were no significant differences in the anti-HBc detection rates for men (31.5%) and women (37.0%). In the structure of seropositive anti-HBc, a smaller percentage was detected in the category “young age” (24.0%), the largest prevalence is noted in the category of “middle age” (45 to 59 years), with 38.8%, which may be associated with service duration and longer exposure to potential HBV sources. The detection frequencies of anti-HBc to HBV nuclear antigen, along with anti-virus surface protein antibodies (anti-HBs) represent an index of HBV prevalence. We would like to recommend anti-HB testing for the medical staff members before repeated immunization and with additional examinations (search for viral DNA with a positive result) in order to exclude medical workers as a source of infection followed by further monitoring of these persons for timely medical aid, as well as adjusting the vaccination volume against HBV in this professional cohort.

Cellular contents of red bone marrow is presented by an extremely heterogeneous cell population including stem cells, reticular cells, and differentiationg cells of five hematopoietic lineages. The current task for cell therapy and experimental studies is to obtain cell fractions of bone marrow enriched with a certain type of cells. In this paper we investigated the level of cytokine mRNA expression in bone marrow cell fractions isolated by counterflow centrifugation in an elutriator rotor. The marrow cell fractions were isolated at a rotor speed of 2500 rpm. Six cell fractions (F-1 to F-6) were collected: F-1, at a buffer flow rate of 12 mL/min; F-2, at 15 mL/min; F-3, at 19 mL/min; F-4, at 23 mL/min; F-5, at 50 mL/min. Fraction 6 was collected after stopping the rotation. Cytomorphological analysis of the fractions showed that erythrocytes (80%) and lymphocytes (40%) are collected in the “light” fraction F-1, lymphocytes (44%), polychromatophilic (50%) and oxyphilic (51%) normocytes – in F-2, neutrophils (70%) and eosinophilic granulocytes (40%) – in F-3 and F-4, macrophages (64%), megakaryocytes (95%), reticular (35%) and mast cells (62%) – in F-6. Blast cells of different hematopoietic lineages were detected mainly in F-5. Using RT-PCR, the maximum gene expression of the stem cell factor (Scf) and granulocyte-macrophage colony-stimulating factor (Gm-csf) was detected in the “heavy” fraction F-6, gene expression of tumor necrosis factor-α (Tnfα) and erythropoietin (Epo) – in F-4, F-5 and F-6, and gene expression of macrophage colony-stimulating factor (M-csf) – in F-3 and F-4. Thus, this method allows to separate the “light” fractions of lymphocytes and erythrocytes from the bulk of bone marrow cells, which can be used in allogeneic bone marrow cell transplantation to reduce the risk of acute graft-versus-host disease. Another important advantage of the method is the ability to obtain fractions of “heavy” cells with high regenerative potential in order to use them in cell therapy in order to stimulate regenerative processes in organs and tissues.

The ability of cytokine-induced killer (CIK) cells to recognize and lyse tumor cells has been shown in preclinical in vitro studies. The database ClinicalTrials.gov currently registers over 600 completed clinical studies on cellular immunotherapy including more than 30 trials with CIK cells. There are many ways to obtain the lymphocyte-based cell products. In our opinion, their common drawback is the need for additional blood samples to obtain cell preparation in large volumes, thus requiring the development of new approaches in the field. We have proposed an idea of lymphocyte extraction using leukocyte filters with subsequent in vitro activation of these cells. The purpose of our study was to test the opportunity of using leukocyte filters as a source of lymphocytes to obtain large doses of CIK. The study included data from 30 donors. The article describes in detail the procedure for extracting lymphocytes from leukocyte filters. The isolated lymphocytes were cultured in plastic bottles in the CO₂-incubator for 10-14 days in a medium based on RPMI-1640 supplied with IL-2 and IL-15. The level of cell activation was assessed using enzyme-linked immunosorbent assay (ELISA) and flow cytometry. The subpopulation profile of T, NK, TNK, B lymphocytes (CD3, CD4, CD8, CD14, CD16, CD19, CD45, CD56) and surface activation markers (CD38 and HLA-DR) before and after cell activation were assessed. We have assessed the possibility of using leukocyte filters as a tool of cell separation from leukocyte-rich plasma. The functional activity of extracted lymphocytes and cytokine contents of supernatants were studied after cultivation in a complete nutrient medium. The supernatant peptide complex was shown to contain high doses of signaling proteins, i.e., IL-2, IL-6 IL-10, IFNγ, TNFα and retained its functional activity after long-term storage at low temperatures. The resulting cell product is sterile and consists predominantly of T, TNK and NK cells with a high level of viability and expressed activation markers (HLA-DR, CD38). Each single filter procedure allowed getting of up to 350 million activated lymphocytes and supernatant aliquots. This preparation may be used both for basic research and for administering autologous immune therapy to cancer patients in order to restore the patients’ own antitumor immunity potential. This amount of cellular and cell-free products is sufficient to conduct more courses of immune therapy without additional blood sampling.

Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease that predominantly affects small joints, causing persistent pain, functional impairment, and a marked reduction in patients’ quality of life. The pathological process is characterized by ongoing synovial inflammation, destruction of cartilage and subchondral bone, and extra-articular manifestations involving the cardiovascular, pulmonary, and nervous systems. A key pathogenetic factor seems to be an imbalance between pro- and anti-inflammatory mediators, among which tumour necrosis factor-a (TNFa), interleukins IL-17A and IL-17F, and osteoprotegerin (TNFRSF11B), a regulator of osteoclast differentiation, play central roles. The present study aimed to assess the contribution of polymorphisms in the TNFA, IL17A, IL17F, and TNFRSF11B genes to individual susceptibility to RA in the Russian population of the Chelyabinsk Region. We hypothesized that the major genetic impact on disease development may be produced not only by single nucleotide variants but also by their combined multilocus constellations. Special attention was given to intergenic interactions, which are often underestimated in most association studies. Genotyping was performed by polymerase chain reaction (PCR). To evaluate the data, we applied the multifactor dimensionality reduction (MDR) algorithm, which constructs predictive case–control models and evaluates their robustness by means of ten-fold cross-validation and permutation testing. The algorithm identified three most informative combinations comprising four to six SNPs each; every combination showed statistical significance and high predictive accuracy. Cross-validation consistency values exceeded 9/10, indicating excellent reproducibility of the models. These findings confirm that a comprehensive multilocus genotype analysis is more informative than examining individual markers alone and it may be used for patient risk stratification, early diagnosis, and development of personalised preventive strategies based on targeted anti-cytokine therapies. Further studies in larger cohorts are needed to validate these results.

Altered immune response in abdominal sepsis (AS) leads to dysregulation of innate and adaptive immunity, followed by immunosuppression. Mortality rates in AS are quite high, thus requiring continued indepth study of the role of adaptive immunity in this condition. The aim of our study was to evaluate the role of changes in adaptive immunity as a factor of an unfavorable AS outcomes. The control groups consisted of 63 conditionally healthy individuals (33 women and 30 men, group 1). 64 patients with AS aged 32-82 years were also observed. According to their clinical outcomes, we discerned group 2 (n = 46) with a favorable outcome and group 3 (n = 18) with a fatal outcome. The studies were conducted on days 1, 3, and 7. The total number of leukocytes and lymphocytes was determined by the Sysmex XT-1800i/XT-2000i hematology analyzer (Japan). The subpopulation composition of lymphocytes was studied in peripheral blood by flow cytofluorometry using Navios 2/6 flow cytometer (Beckman Coulter, USA) with monoclonal antibody conjugates (Beckman Coulter, USA). The concentrations of IgA, IgM, and IgG were determined by solid-phase enzyme immunoassay (JSC "Vector-Best", Russia). The contents of circulating immune complexes were determined with a microplate reader (Labsystems Multiskan Plus, Finland) using the CIK-HEMA test systems (Hema-Medica, Russia). Statistical processing of the results was performed using the statistical package SPSS 17.0. Analysis of adaptive immunity cells in patients with AS revealed lymphocytopenia: deficiency of CD45⁺CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺, immunoregulatory index, CD3^-CD19⁺, marker of early activation of T lymphocytes (CD3⁺CD25⁺). A significant relationship was found between the severity of leukocytosis, lymphocytopenia and the outcome of the disease. In cases of favorable outcome, lymphocytopenia was less pronounced than among patients with unfavorable one, along with significantly higher values of the lymphocyte subpopulations: CD45⁺CD3⁺, CD3⁺CD4⁺, immunoregulatory index, CD3^-CD19⁺, CD3⁺CD25⁺, CD3⁺56⁺. Meanwhile, the number of CD3⁺CD8⁺, CD3⁺HLA-DR⁺ lymphocytes in the surviving patients was decreased over 7 days, as compared with the unfavorable outcome group. More pronounced leukocytosis was associated with unfavorable outcome of AS, thus emphasizing severity of the systemic inflammatory response, which is closely associated with severity of clinical course and prognosis of AS. There was a more pronounced lymphocytopenia, a progressive decrease in the following lymphocyte subpopulations compared to the survivors: CD45⁺CD3⁺, CD3⁺CD4⁺, immunoregulatory index, CD3^-CD19⁺, CD3⁺CD25⁺, CD3⁺56⁺. The values of CD3⁺CD8⁺, CD3⁺HLA-DR⁺ lymphocytes in fatal cases were higher than in favorable cases. The results of our study emphasize the need to take into account the state of adaptive immunity in order to more accurately stratify the risk of death and personalize therapeutic strategies.

Dental implants are widely used in current dentistry and effectively replace the lost teeth. However, due to concomitant risk factors and low oral immunity, the patients develop peri-implantitis (PI), accompanied by inflammation and general immune response. Meanwhile, the local cytokine profile in saliva remains insufficiently studied. The purpose of this study was to assess cytokine response and evaluate the diagnostic information content of proand anti-inflammatory cytokines in patients with PI. We have examined sixty-five patients with PI aged 50-65 years and 48 PI-free patients aged 50-65 years. The PI diagnosis was carried out based on the results of clinical examination, presence of symptoms, i.e., bleeding during probing, suppuration during probing, bone loss on radiographs ≥ 3 mm, and the depth of the gingival pocket ≥ 6 mm. In saliva samples collected in the morning on rest, the content of several proand anti-inflammatory cytokines was determined by enzyme immunoassay (ELISA) using Protein Contour kits (St. Petersburg). The cytokine profile of patients with PI is characterized by expression of both pro-inflammatory and anti-inflammatory cytokines. The salivary content of IL-1 increased, particularly, among patients with PI (205.9±4.2 pg/mL versus 67.5±3.1 pg/mL in participants without peri-implantitis). IL-6 levels were increased to 29.4±1.3pg/mL versus 11.7±1.2 pg/mL, respectively, showing a significant difference for the both parameters. In addition, the level of IL-19 increased to 54.9±2.3 pg/mL against the comparison group (30.5±1.4 pg/mL); TNFα, to 202.4±3.8 pg/mL compared to 115.6±2.4 pg/mL, respectively (p < 0.001). Moreover, the salivary concentration of IL-4 was significantly increased in patients with PI (18.2±1.4 pg/mL versus 3.8±0.6 pg/mL in comparison group), along with higher IL-10 amounts (66.5±2.7 pg/mL versus 15.3±0.9 pg/mL, respectively, p < 0,001). An estimation by the Kullback information measure revealed the highest diagnostic information content for IL-4 (8.5), IL-10 (8.1), IL-1β (5.7), IL-6 (4.2). To justify the diagnostic criteria, one may recommend usage of IL-4, IL-10, IL-1β and IL-6 levels established in saliva.

Usage of psychoactive substances may cause toxic encephalopathy, which contributes to the modulation of both innate and adaptive immune responses. The impact of psychoactive substances, including those illegal in Russia, upon immune homeostasis remains poorly studied, especially in northern regions with extreme climatic factors. With regard to medical and social consequences, attention is drawn to the rapidly increasing social maladaptation of young patients, as well as the manifestations of organic brain damage. This study is aimed at assessing the ratio of lymphocyte proliferation and apoptosis events in male subjects living in the Northern region of Russia suffering with toxic encephalopathy caused by psychoactive substances. The work included a comparative analysis of immunological parameters of peripheral blood determined in two groups of people living in the Northern region of the Russian Federation, i.e., twenty men with toxic encephalopathy (mean age 33.8±1.8 years) on the first day after severe poisoning with psychoactive substances versus 22 healthy volunteers (34.5±2.0 years, control group). Phenotyping of peripheral blood lymphocytes was performed by the method of indirect immunoperoxidase reaction with monoclonal antibodies. Statistical data processing was performed using SPSS 25.0. The results revealed a statistically significant (p < 0.01) increase in lymphocytes with the CD10⁺ marker in abusers of psychoactive substances (0.54 (0.40-0.75) × 10⁹ cells/L) compared to the control (0.27 (0.16-0.51) × 10⁹ cells/L), which indicates hyperproliferation of lymphocytes due to CD10⁺. At the same time, a low number of lymphocytes with the CD95⁺ marker was recorded in men with psychoactive substances abuse compared to the control group, thus reflecting the suppression of apoptosis. A two-fold increase in the CD10⁺/CD95⁺ ratio was established in men with psychoactive substances poisoning, demonstrating an imbalance in proliferative-apoptotic processes. A distinctive feature for men with toxic encephalopathy is an increased content of CD10⁺ cells that stimulate immune responses via the classical pathway, along with low concentration of lymphocytes with the CD95⁺ marker, which indicates an increased expenditure of the reserve capacity of immune homeostasis and their reduction. We have also revealed a moderate decrease in the levels of CD71 receptor expression on lymphocytes in males with psychoactive substances abuse. The data obtained indicate that in individuals with toxic encephalopathy in Northern Region, a disruption of both the long-term and short-term mechanisms for maintaining immune homeostasis is observed, being associated with an imbalanced ratio of lymphoproliferation and apoptosis processes.

Some patients with inflammatory bowel disease (IBD) may lack the conventional inflammation criteria, i.e., elevated CRP, ESR and leukocytosis. The aim of this study was to search for additional laboratory risk criteria of IBD in patients with non-specific inflammation markers. The study involved 150 patients divided in two groups: an observation group (100 patients with a verified diagnosis of ulcerative colitis or Crohn’s disease) and a control group (50 clinically healthy individuals). All subjects were tested for complete blood analysis, blood biochemistry. IL-1β, TNFα, and IL-4 concentrations were also assayed. As based on general blood counts and biochemistry testing, the patients from the observation group were divided in two subgroups, i.e., the persons with classic laboratory signs of inflammation, and those without detectable inflammation markers. Three main laboratory blood parameters were assessed for diagnostics of inflammatory process: ESR, white blood cell counts and CRP level. Increase in two or more of the above blood parameters had been considered a positive finding. It was noted that 40% of patients with IBD lacked the non-specific laboratory criteria of inflammation: in 37% of patients with ulcerative colitis, and in 46% of Crohn’s disease cases (p < 0.001). Moreover, we carried out a comparative analysis of cytokine levels and biochemical markers in the blood serum from the controls, and IBD patients without laboratory signs of inflammation. Based on the obtained data, we developed a prognostic model for the IBD probability in patients without non-specific inflammation markers, depending on biochemical and immunological blood parameters. The model included such serum parameters as glucose, sodium and IL-4 concentrations. The predictive ability of the model was assessed using ROC analysis (AUC 0.970±0.018 95% CI: 0.936-1.000; p < 0.001). An algorithm for predicting the IBD risk in patients without non-specific laboratory criteria of inflammation was proposed. The obtained data enable us to identify additional criteria for the IBD risk in patients lacking the non-specific metabolic criteria of inflammation.

The mechanisms of liver regeneration after damage are associated with activation of cytokines. SCF is an important signaling factor since, along with hematopoiesis regulation, it is involved in liver restoration and regeneration after damage. In this respect, the aim of our study was to evaluate SCF levels in blood plasma and liver during physical activity of varying intensity and after Meldonium admonistration. Material and methods: The experiments were conducted with male rats. The study was performed in two stages. Initially, the rats were subject to exertion of varying intensity: the 1^st group of rats performed light physical activity, the 2^nd group underwent moderate exercises, the 3^rd group performed heavy physical activity. A total of 10 sessions of swimming loading were performed. The Intact, sexually mature male rats did not undergo physical activity. The number of these animals corresponded to the number of experimental group. The rat SCF concentration (rat stem cell factor / mast cell growth factor) in blood plasma and the levels of SCF in the liver tissue were determined by means of enzyme immunoassay. The study has shown that SCF level in the intact group was higher in liver tissue than in blood plasma. In the 1^st group of animals, an increase in SCF by 1.5 times in the blood and a decrease in the liver were noted, as compared to the intact group (p < 0.05). In the 2^nd group, SCF contents in the liver was increased, along with a decrease in blood plasma by 1.1 times when compared to the 1^st group. However, this parameter remained higher than in intact animals by 1.4 times (p < 0.05). In the 3^rd group, a decrease in SCF was noted both in blood plasma, and in liver samples as compared to intact animals (p < 0.05). After administration of meldonium during mild and moderate physical exertion, an increase in SCF in the liver by 1.1 times was noted. During severe physical exertion it was 1.4 times higher when compared to experimental animals that did not receive meldonium. Hence, usage of meldonium during physical exercise of varying intensity led to a significant increase in the liver SCF concentration.

Atopic dermatitis is a chronic, relapsing inflammatory skin disease. Immune dysregulation occurs, leading to inflammation in response to allergens that penetrate through a compromised epidermal barrier. Defects in the skin barrier and inflammatory processes in the skin result in increased colonization by conditionally pathogenic microorganisms. Fungal microbiota dysbiosis (Malassezia spp. and others) is considered one of the factors involved in the pathogenesis of atopic dermatitis. However, the influence of fungal microflora on atopic dermatitis remains less studied compared to the role of bacterial microorganisms. The aim of this study was to conduct a comparative analysis of the prevalence of fungi in patients with atopic dermatitis and a control group using real-time polymerase chain reaction. The study included 151 participants. The group of patients with atopic dermatitis included lesions from different locations, and for more accurate comparison. The control group was formed considering both dry skin areas and areas with increased sebum production. All patients had their diagnosis of atopic dermatitis confirmed according to clinical guidelines. Disease severity was assessed using the SCORAD index. Epithelial scrapings were collected depending on lesion location. The obtained skin samples were analyzed using real-time PCR. It was found that patients with atopic dermatitis exhibited significantly higher levels of fungal colonization compared to the control group. Specifically, patients with atopic dermatitis more frequently harbored various yeast species such as Candida, Malassezia, and Saccharomyces cerevisiae, as well as non-dermatophyte molds like Aspergillus spp./Penicillium spp. Additionally, we observed that the distribution of Candida and Malassezia restricta species varies depending on the severity of atopic dermatitis. Changes in the fungal composition of the skin play an important role in the pathogenesis of atopic dermatitis, potentially activating the immune system and exacerbating inflammatory processes. Although fungi are not direct causative agents of atopic dermatitis, an imbalance in their populations likely contributes to disease exacerbation and progression. Further research is needed to better understand how fungi influence atopic dermatitis development particularly studies focusing on immune responses to fungi and allergy mechanisms related to their alterations. Such data could aid in developing new therapeutic and preventive strategies for this condition.

The ecologically unfavorable Trans-Aral region is characterized by high prevalence of chronic respiratory allergies in children, often combined with anemias. These disorders significantly impact children’s health and require comprehensive study of immune and physiological features. Our objective was to assess the characteristics of cytokine profile in children from different age groups with respiratory allergies accomplished by anemia, depending on the presence of concomitant dermatoses, and level of physical development. The study included 284 children aged 3 to 17 years, divided into three groups: 121 children with respiratory allergies (RA), 126 children with respiratory allergies and concomitant dermatoses (RA + D), and 37 practically healthy children (control group). Concentrations of key cytokines (IL-4, IL-6, IL-10, IL-18, TNFα), and total IgE in blood serum samples were determined by enzyme-linked immunosorbent assay (ELISA). Physical development was assessed using the standardized WHO AnthroPlus methodology. Children with respiratory allergies, regardless of the presence of dermatoses, showed significantly increased levels of IL-4 and IgE compared to controls (p < 0.01), thus suggesting emergence of a dominant Th2 response. The highest level of anti-inflammatory cytokine IL-10 was found in the RA + D group (p < 0.01), likely reflecting a compensatory mechanism aimed at limiting inflammation. Meanwhile, pro-inflammatory cytokines IL-6 and TNFα were significantly decreased in patients compared to healthy children (p = 0.001 and p < 0.001, respectively), thus suggesting probable hyporesponsiveness of the pro-inflammatory pathway in chronic allergic conditions. The levels of IL-4 and IL-10 proved to be lower in children with delayed physical development, while IL-18 was relatively increased, being consistent with literature data and suggesting possible mechanisms of immune dysregulation associated with impaired general health status. The data obtained suggest a predominance of Th2 immune response in children from Trans-Aral Region with respiratory allergies as reflected by increased IL-4, IL-10, and IgE levels. The decreased IL-6 and TNFα levels warrant further study to understand inflammatory processes in chronic allergies under the conditions of ecological stress. Accounting for cytokine profiles and physical development is important for development of personalized therapeutic approaches and improving the disease prognosis.

Adoptive CAR-T cell therapy is an innovative approach in oncology that uses genetically modified autologous T cells from the patient as a therapeutic tool to fight cancer. The use of DNA plasmids and in vitro transcribed (IVT) mRNA as vectors for the production of CAR-T lymphocytes has a number of advantages compared to viral vectors, such as the absence of cell genome modification, high transfection efficiency, speed and potentially lower cost of obtaining the final product. In current work, we studied the efficiency of transfection (in terms of cell viability and expression of the target protein) of peripheral blood mononuclear cells and cells of transplanted culture (human embryonic kidney cells, HEK293) by electroporation using model DNA plasmid (pmaxGFP) and IVT-mRNA (mRNA-GFP) encoding green fluorescent protein (green fluorescent protein, GFP). The selection of an optimal transfection regimen was performed. It has been shown that although mRNA-GFP yields a comparable number of cells GFP-expressing cells, the cell viability, and, consequently, general efficiency of transfection is significantly higher when using mRNA-GFP as a vector. At the same time, a comparison of expression level by the cells transfected by two techniques showed that the use of mRNA provides more uniform parameters, whereas usage of plasmid vector results in expression levels differing by several orders of magnitude. The changes in expression level were also tested within 7 days after transfection. It was shown that the proportion of GFP-positive cells decreases with time and does not depend on the method of transfection, while the assessment of the proportion of viable cells showed that plasmid transfection leads to a decreased proportion of viable cells after 7 days to 30%, while the use of mRNA practically does not affect viability (the number of viable cells after 7 days did not significantly differ from the control). The results obtained indicate that the usage of IVT-mRNA may be a more preferable tool in production of CAR-T products by electroporation.

Pruritus is a rare chronic recurrent polyethological skin disease characterized by severe itching, resulting in development of skin elements such as papulo-vesicles, papules, nodules and plaques. The disease is often difficult to treat with standard therapy, and patients experience debilitating symptoms for years. The article presents a clinical case of a patient who suffered from nodular pruritus for more than 20 years, which worsened over last 4 years after suffering from a mild COVID-19 infection, when the disease became continuously recurrent. Conventional treatment was virtually ineffective. Skin rashes became widespread upon admission to Clinic of Immunopathology at the Research Institute of Fundamental and Clinical Immunology (Novosibirsk) in April 2024, severe itching disrupted sleep, thus significantly reducing the quality of life. On 04/16/2024, immunotherapy was initiated with dupilumab at initial dose of 600 mg, according to the instructions, followed by 300 mg every 2 weeks. After the first administration of the drug, significant positive dynamics was noted, i.e., decrease in itching, rapid regression of rashes, and the patient’s skin was almost completely cleansed by the 4th injection. At the present time, the skin remains clean, there is no itching, and sleep has returned to normal. The patient continues to apply emollients and takes second-generation antihistamine drugs. The efficiency of dupilumab therapy, which blocks IL-4 and IL-13 signaling by specifically binding to IL-4Rα, a subunit common to IL-4 and IL-13 receptor complexes, is due to modulation of neuroimmune regulation in the skin which underlies pathogenesis of chronic itching in nodular pruritus. IL-4 and IL-13 are considered important cytokines of Th2 cells, which increase the expression of IL-31R and other pruritus-related receptors, enhance neuronal activation in inflamed skin, transmitting signals to nearby efferent neurons and the central nervous system, thus causing itching. A vicious cycle of itch-scratching is formed, with enhanced sensitization of histamine-independent sensory neurons to pruritogens, enhanced fibroblast proliferation, migration, and production of profibrous factors. Since the main goal of treating nodular pruritus is to block the vicious cycle of itching and scratching, and promote nodular healing, dupilumab therapy leads to remission of the disease, normalizing the interaction of immune and nervous systems in the skin. When treating adult patients with nodular pruritus, dupilumab was recently approved in Russia (just a year ago), and has already shown its high efficiency. The presented case demonstrates the opportunities of modern immunotherapy methods in treatment of dermatological diseases with sluggish clinical course.