Endometriosis is a chronic recurrent disease with insufficiently studied pathogenesis. Endometriosis is known to share similar features with tumors. Thus, the outgrowth of endometrium-like tissue outside the uterus is the main feature of this condition. The dysfunction of local immune response is required for cell proliferation and invasion in ectopic sites. The involvement of immune checkpoints is among the mechanisms allowing avoidance of immune surveillance shown for the tumors. Immune checkpoints are presented by proteins expressed on immune cells (most on T cells). The checkpoint binding to its ligand expressed on immune cells leads to its functional inhibition and, thus, facilitates survival of tumor cells. The data about immune checkpoints, e.g., CTLA-4, PD-1, LAG-3, Tim-3, TIGIT, 4-1BB, GITR are summarized in this review. Their ligands (CD80/CD86, PD-1L, Gal-3, Gal-9) are also described. The review article contains information about cells expressing checkpoints and other proteins involved, We also discuss the examples of tumors using such checkpoint-ligand interactions in order to avoid recognition by immune cells. Furthermore, the review describes immune checkpoint inhibitors currently used in cancer therapy. Due to scarce knowledge about endometriosis pathogenesis, the only diagnostics of this condition is laparoscopic surgery with visualization of ectopic loci and histological study of biopsies. The studies of some biomarkers for non-invasive diagnosis of endometriosis, such as CA-125, MCP-1, IL-6, BDNF etc. are also discussed in this review. The authors describe some studies which concern immune checkpoints in the context of endometriosis. Noteworthy an elevated expression of some checkpoints by T cells was found, along with elevated concentration of their soluble forms in blood of women with endometriosis. The review also includes the studies showing significant sensitivity and specificity of immune checkpoint ligand measurement in patients with endometriosis. Hence, investigation of immune checkpoints as a potential mechanism to avoid immune reaction used by endometriotic cells, and its application as a biomarker for non-invasive diagnostics is a promising direction for the further studies. 

Lipopolysaccharide (LPS, endotoxin) of Gram-negative bacteria is a strong activator of innate immune system and inducer of systemic and local inflammation. Due to increasing number of factors contributing to the translocation of LPS into the systemic bloodstream, e.g., non-adequate antibiotic therapy, usage of entero- and hepatotoxic drugs, as well as increased proportion of carbohydrate and fatty foods in the diet of modern people, the role of LPS is growing, in view of maintaining low-grade inflammatory background. Interactions of endotoxin within human body are mediated by a number of receptors and carrier molecules, many of which can be distinguished into a group of so-called “LPS-binding systems”, i.e., lipopolysaccharidebinding protein (LBP) and bactericidal/permeability-increasing protein (BPI). The character of response to increased LPS pool in blood circulation depends largely on these molecules, as well as additional substances that interact with LPS and LPS-binding systems, in particular, low-density lipoproteins (LDL) and high-density lipoproteins (HDL). Given current publications reporting elevated LPS levels in patients with rheumatoid arthritis (RA), and persistence of dyslipidemias in the vast majority of these patients, LPS is potentially a pathogenetically important factor in RA. This review presents basic data on the biology and role of LPS and “lipopolysaccharide-binding systems” in development and maintenance of inflammation state in RA. Information was searched using the keywords “rheumatoid arthritis and lipopolysaccharide”, “rheumatoid arthritis and lipopolysaccharide-binding protein”, “rheumatoid arthritis and BPI” in foreign and Russian scientific databases, including e-Library and PubMed. The presented data allow us to consider the combination of “lipopolysaccharide-binding systems” imbalance and dyslipidemia a sufficient aggravating pro-inflammatory factor in RA, and the search for potential mechanisms influencing these conditions, either separately, or in combined manner, as a promising field for clinical research.

Chronic rhinosinusitis (CRS) is a disease which manifests with inflammation of the upper respiratory tract. Two main phenotypes can be distinguished in CRS: a clinical form with polypous tissue, and a clinical variant without polyposis. With regard of increased cytokine concentrations, the inflammatory response in CRS can be divided into 3 endotypes: Th1 (IFNγ), Th2 (IL-4, IL-5, IL-13) and Th3 type (IL-17, IL-22). The pathogenesis of inflammation in CRS with nasal polyps and polyposis-free cases is quite different, and, according to current publications, the data on prevalence of different endotypes is very contradictory, thus confirming the need for further studies of CRS development. These important medical and social features of diseases affecting nasal mucosa and paranasal sinuses require further studies in pathogenesis of CRS. This review covers information about the immunological features and dysfunctions that lead to occurence of CRS with or without polyps. The purpose of this review article is to study the influence of the first-line immune defense, components of innate and acquired immunity on the pathogenesis of CRS.
The article provides a review of the worldwide research publications in the field. The authors conducted a search for different items of immune response related to development of CRS with and without polyps. We used keywords and filters in the PubMed and Google Scholar, as well as in Scopus and Web of Science databases.
So far, low efficiency of various treatment methods used may be due to heterogeneous immunopathology. The use of biological preparations, although approved, may be non-reliable, since these Th2-targeted drugs may be administered to patients with non-Th2 disease. The presence of eosinophils and pus may provide a basis for endotype extrapolation. However, the clinicians treating CRS do not have widespread access to laboratory tests in order to specify the CRS type and to administer a tailored drug management. Patients with any type of inflammation may suffer from latent infections caused by bacteria, fungi or viruses, thus making difficult a specific evaluation of polarized immune response. Further studies on the links of immunological pathogenesis in CRS will allow us to develop a personalized algorithm for the diagnosis and treatment of such patients.

The review article is devoted to the two key Th2 cytokines, IL-4 and IL-13, which are directly involved in the immunopathogenesis of atopic dermatitis (AD). The identification of IL-4 and IL-13 in AD was first reported by Q. Hamid et al. in 1994. Since then, a number of studies have appeared confirming the relationship of these Th2 cytokines with disruption of the skin epidermal barrier; a decrease in skin immune response due to inhibited expression of antimicrobial peptides against Staphylococcus aureus, etc. The convincing studies also confirm a relationship with IL-4/IL-13 to such clinical manifestations of ADs as skin infections, as well as inflammation, lichenification and itching of the skin. The role of IL-4 and IL-13 is also confirmed by clinical studies, which indicate a beneficial effect of drugs inhibiting these cytokines on the relief of skin symptoms in atopic dermatitis (itching, rashes). The IL-4 and IL-13 are shown to connect the JAK/STAT signaling pathway due to the common α-subunit of IL-4 receptor (IL-4Rα). Importantly, IL-4, IL-13 and other cytokines (including IL-31) are capable of activating the sensory neurons, thus being often considered potent pruritogens. The article also discusses issues related to the role of the JAK/STAT signaling pathway and, in particular, the JAK1 protein in development of atopic dermatitis. As based on pathogenetic significance of IL-4 and IL-13, drugs have recently been developed that block their activity and, thereby, affect important molecular pathways of the AD development. These drugs are classified as systemic medications which include, e.g., (1) biological therapy (dupilumab, the first monoclonal IgG4 antibody), which blocks IL-4Rα and, thereby, suppresses the IL-4/IL-13 axis, and (2). Janus kinase (JAK) inhibitors or small-molecule agents. Currently, some Janus kinase inhibitors, e.g., abrocitinib, upadacitinib, and barocitinib, are available in Russia. Clinical studies show that both biological therapy and small molecules have an immunomodulatory effect on the course of atopic dermatitis. The review briefly presents the main data of recent meta-analyses on the comparative characteristics of biotherapy and usage of Janus kinase inhibitors in this disorder.

The development of new effective methods for in vitro activation and culturing of natural killer (NK) cells led to advances in adoptive cancer immunotherapy. Our aim was to evaluate expression of activation markers on natural killer cells after cultivation and activation in vitro using flow cytometry. Peripheral blood mononuclear cells were isolated from heparinized blood of 10 donors in a density gradient and were cultivated in RPMI-1640 medium (Sigma-Aldrich, Germany) supplied with IL-2 and TMDK562-15 feeder cells for 14 days. Phenotyping of the NK cells was carried out using an Image Stream MkII (Luminex) flow cytometer. We used a cytometry panel containing 10 monoclonal antibodies against СD45, СD3, СD56, СD16, and against cell activation markers, i.e., CD38/HLA-DR/CD25 (BD Biosciences). Cytotoxic activity of the donor-derived NK cells was tested towards cancer cell lines AsPC-1, A-549, MDA-MB-231 and PC-3 on 14th day of cultivation. The effector-to-target ratios were 5:1 and 10:1. The target cells were exposed to effector cells for 12 hours. Statistical analysis was performed using MS Excel 2016, TIBCO STATISTICA 13. Prior to cultivation, the percentage of natural killer cells in mononuclear cells was 12%. Of those, activation markers (CD38/HLA-DR/CD25) were expressed, accordingly, on 7.6%, 0.7%, 0% cells, Cultivation of mononuclear cells for 14 days resulted in activation and expansion of distinct lymphocyte subpopulations. NK cell counts increased 390-fold on 14th day of cultivation, with percentage of natural killers reaching 86.5%. On the 14th day of cultivation, expression levels of activation markers (CD38/HLA-DR/CD25) were 34.7%, 36.5%, 5%, accordingly. Studying the cytotoxic activity of NK cells against different cancer cell lines showed that, after 12-h incubation of activated cells with target cells, 100% of cancer cells have been lysed. The donor cells were viable by 98%. After 7 days of cultivation, the cell viability decreased to 81%; at the end of cultures, their viability was 95%. The suggested cultivation technique for mononuclear cells cause an increase of proliferative activity and expression of activation markers of NK cells, thus allowing to maintain high viability of the cell population. High functional activity of cultured NK cells was confirmed in cytotoxicity tests against cancer cells.

Human mononuclear cells from umbilical cord blood (HUCBMCs) are used as the main or adjuvant therapy for treatment of about 80 different diseases, due to high proliferative activity of these cells, low immunogenicity and an opportunity of selecting rare HLA types for transplants. In this regard, assessment of cellular material in protocols of immunopharmacology is relevant. Our objective was to study allergenic and immunotoxic effects of mononuclear cells from human umbilical cord/placental blood as preclinical testing in laboratory animals. The study of type I hypersensitivity to HUCBMCs was carried out using a standard method for assessing bronchiolar spasm in male and female guinea pigs. The samples of tracheal sections were incubated in Ringer–Tyrode solution at the 2.5 per cent concentration of mononuclear cell suspension, with histamine hydrochloride serving a positive control. Antibody detection to HUCBMCs was carried out in the CBAxC57B2/6 male mice using by means of complement fixation test (indexed as absence of hemolysis of sheep erythrocytes). The mice were subjected to single intravenous injections of cell material exceeding the human therapeutic dose 10, 50 and 100-times (8.57 × 10⁷  cells/kg, 4.28 × 10⁸  cells/kg, 8.57 × 10⁸  cells/kg body weight, respectively). Blood for analysis was taken 21 days after administration of the biomaterial. Blood serum from mice immunized with S. aureus was used as a positive control. A study of the phagocytic activity of neutrophils was carried out in male and female Wistar rats, which were subjected to a single intravenous injection of HUCBMCs at 10-fold therapeutic dosage. After 30 days, the phagocytic index and phagocytic number were studied using the ink test method, by analyzing 600 cells for each group. The median, upper and lower quartiles (Me (Q_0.25-Q_0.75)) were calculated; the hypotheses were compared using the Mann–Whitney U test. We didn’t detect anaphylactogenic activity and production of antibodies to cellular material after administration of HUCBMCs to the animals. In female rats, the phagocytic activity of neutrophils increased statistically significantly (p = 0.004) relative to control animals [56.5 (53.8-60.8) versus 44.0 (40.5-47.5), respectively]. In male rats, there was a tendency to increased phagocytic activity by 13% (p = 0.054). The phagocytic index in all compared groups remained within deviations of standard values (1.8-2.0). Human umbilical cord blood mononuclear cells do not exhibit an anaphylactogenic, and do not show any immunotoxic effect at 100-fold at the human therapeutic dosage (8.57 × 10⁸  cells/kg). However, they contribute to increase of phagocytic activity of neutrophils, thus requiring further preclinical and clinical trials of efficiency and safety for usage of this biomaterial with high therapeutic potential.

For millennia, the mankind has been trying to fight the causative agent of tuberculosis, Mycobacterium tuberculosis, but morbidity and mortality rates, unfortunately, remain high all over the world. The purpose of our work was to specify changes of immunohistochemical parameters in different areas of the lungs in stable tuberculosis. We have studied 30 cases of stable tuberculosis in male patients. The study concerned two 2 areas: the tuberculoma capsule and adjacent areas (perifocal zones). The comparison group consisted of resected material from upper lobes of the right lung, taken from 10 men who died in accidents and did not have tuberculosis during their lifetime. This group was called “conditionally healthy” controls. To assess the signs of local immune response, an immunohistochemical method was used using monoclonal antibodies to CD8, CD4, CD79a, Fascin. When assessing location of the markers expressed in the lung tissues of “conditionally healthy” group, the number of CD4⁺ and CD8⁺ cells was determined mainly in the interstitial space. The numbers of CD4⁺ cells in this study group prevailed over CD8⁺ cells, whereas CD79a⁺ and Fascin⁺ cells were found in small quantities within interstitial space. In stable tuberculoma, a greater number of lymphocytes were detected in the capsule with a predominance of T killers over T helpers. In perifocal area of stable tuberculoma, the number of immune cells was reduced when compared to the “conditionally healthy” group. At the same time, we determined the areas where CD4-positive lymphocytes formed contacts with interstitial macrophages. The number of CD79a positive cells was mainly determined in the connective tissue capsule of tuberculoma, with single cells on the periphery, being close to macrophages. Fascin-positive cells in both capsule and perifocal zone of stable tuberculoma were rare, being mainly localized around blood vessels. When studying correlations in the “conditionally healthy” group, a moderate positive relationship between cytotoxic lymphocytes and T helper cells was revealed. In the capsular area of stable tuberculoma, strong positive correlations were found for CD4⁺/CD79a and CD79a/CD8⁺ pairs; in the peripheral zone of the lungs, correlations were revealed only between CD79a and CD8⁺ cells. Hence, the degree of activity of the tuberculosis process in the capsule and perifocal areas of stable pulmonary tuberculosis is associated with different cellular interactions which exert double control from both cellular and humoral immunity.

Tick-borne encephalitis is a natural endemic disease which is widely spread in Russia. The purpose of the study was to determine clinical significance of HLA class II genes in tick-borne encephalitis. We observed 75 patients with tick-borne encephalitis admitted to the Kirov Hospital of Infectious Diseases and district hospitals over 2020-2023. Molecular typing of the HLA genes DRB1, DQA1 and DQB1 was carried out using PCR technique, with a set of commercial sequence-specific primers (“DNA-Technology”, Russian Federation). The febrile form of tick-borne encephalitis was noted in 41.3% of patients; focal, in 34.7%; meningeal, in 16.0%, inapparent, in 8% of cases. The comparison group for HLA DRB1 locus included 1528 practically healthy individuals from the same population. Comparison group for HLA DQA1 and DQB1 genes comprised 133 persons. The study has revealed a number of HLA class II genes, which are found significantly more often in TBE patients, rather than in control group (DRB1*1 (χ² = 12.2; p_c < 0.01), DRB1*4 (χ² = 6 .4; p_c < 0.05), DRB1*7 (χ² = 11.7; p_c < 0.01), DRB1*8 (χ² = 4.6; p_c < 0.05), DRB1*13 (χ² = 7.7; p_c < 0.01), DRB1*15 (χ² = 9.3; p_c < 0.01), DRB1*16 (χ² = 14.3; p_c < 0.01), DQA1*0102 (χ² = 7.6; p_c < 0.01), DQB1*0401-2 (χ² = 3.9; p_c < 0.05), DQB1*0502-4 (χ² = 8.1; p_c < 0.01). Among HLA class II haplotypes, the susceptibility to the development of tick-borne encephalitis was determined by the combinations DRB1*08-DQA1*0401-DQB1*401/402 (χ² = 5.7; p_c < 0.05), DRB1*09-DQA1*0301- DQB1*303 (χ² = 5.7; p_c < 0.05) and DRB1*16-DQA1*0102-DQB1*502 (χ² = 7.4; p_c < 0.01). Carriage of the DRB1*15 gene was most risky for development of febrile form of tick-borne encephalitis, (χ² = 7.8; p_c < 0.01; RR = 3.1). Occurrence of three-locus haplotypes DRB1*09-DQA1*0301-DQB1*303 (χ² = 8.8; p_c < 0.01), and DRB1*16-DQA1*0102-DQB1*502 (χ² = 5.0; p_c < 0.05) was associated with increased risk of developing a febrile form of TE by 14.5 and 10.9 times, respectively. In patients with meningeal form of EC, compared with healthy individuals, the gene variants DRB1*08 (χ² = 12.9; p_c < 0.01), DQA1*401 (χ² = 3.9; p_c < 0.05), DQB1*401/402 (χ² = 9.1; p_c < 0.01) were significantly more common. The presence of a threelocus haplotype DRB1*16-DQA1*0102-DQB1*502 (χ² = 10.9; p_c < 0.01) increases the risk of developing a focal TBE by 17.7 times. Thus, tick-borne encephalitis is associated with certain HLA class II alleles, which may be used as a prognostic criterion for development of different clinical forms of tick-borne encephalitis, or tick-borne encephalitis in general.

Chronic urticaria is referred to recurrent, pruritic, erythematous, and edematous mucocutaneous lesions on most days of the week, and persists for six weeks or more. There is a hypothesis about the levels of blood lipid profiles in CSU, which may have a contributing role in development or exacerbation of hives attacks. The present study was conducted to investigate the association between chronic urticaria and blood lipid profiles. Fifty patients with chronic urticaria and fifty healthy people were included in this case-control study. In chronic urticaria patients, each parameters of blood lipid profile and urticaria severity were evaluated in each of four age and two sex cathegories. Urticaria severity in chronic urticaria patients, was also analyzed for levels of HDL, LDL, Triglyceride and Cholesterol. Levels of triglyceride (p value = 0.039), total cholesterol (p value = 0.031), and LDL (p value = 0.001) were significantly higher in chronic urticaria patients than in control group. No correlation was found between the urticaria severity (UAS7 score) average, and the age and sex of the patients. Urticaria severity showed no significant difference within each lipid profile parameter. Average values of lipid profiles in patients with chronic urticaria in different age and sex showed that HDL has remarkably higher mean quantitation in women than men (p < 0.002). Our study found a correlation between chronic urticaria with hyperlipidemia. According to this investigation, we can advise that patients with chronic urticaria should be evaluated for hyperlipidemia.

Aim of the work: to evaluate urinary CD163 as a possible biomarker indicating activity of lupus nephritis (LN). This retrospective, cross-sectional study evaluated 68 patients diagnosed with systemic lupus erythematosus (SLE) over a year, focusing on different states of lupus nephritis (LN). Participants included 38 with active LN, 15 with a history of LN in a non-active phase, and 15 without kidney involvement. The study utilized the SLEDAI index to classify disease activity, with active LN identified through specific urinary parameters. Renal biopsies were performed for those with active disease, following established classification criteria. Comprehensive assessments included blood tests, urinary protein levels, and measurement of urinary sCD163 using ELISA. Statistical analyses employed SPSS, utilizing various tests to compare groups and assess relationships between urinary sCD163 levels and clinical characteristics, establishing significance at p < 0.05. The findings contribute to the understanding of renal manifestations in SLE and the potential role of urinary biomarkers in monitoring disease progression and activity. Laboratory data from 68 participants were analyzed, focusing on correlations among active LN, inactive LN, and SLE without renal involvement. Significant correlations (p < 0.05) were observed in CD163, C3, C4, hemoglobin, platelets, serum creatinine, proteinuria, and BUN, while WBC count, serum albumin, and ESR showed no significant correlation. Notably, 98.5% of patients had positive anti-ds-DNA antibodies. Urinary sCD163 levels were highest in active LN patients. Linear regression showed that serum albumin and ESR significantly predicted urinary sCD163 levels. The optimal cut-off for urinary sCD163 to predict renal activity was > 4.2, with 60.5% sensitivity and 66.7% specificity. However, sCD163 levels did not correlate with renal histopathological classifications. Integration of urinary sCD163 as a biological marker for evaluating the activity of LN together with accurately distinguishing between histopathological classes mostly needs to be further evaluated. To this point of the study, sCD163 can be a good indicator of LN activity, sCD163 still can’t substitute for renal biopsy in differentiation of LN classes as it would not provide the comprehensive understanding necessary for effective management of LN.

A number of studies have shown an association of gastritis symptoms with the levels of specific cytokines. However, some data observed are quite contradictory. Meanwhile, the peptic ulcer disease is known to be associated with unfavorable course of inflammatory processes in gastric mucosa, Moreover, the potential relationships between gastritis and blood cytokine levels in cases of familial predisposal to gastric ulcer have not been studied in children’s age. The aim of our study was to evaluate associations of cytokines in blood serum (IL-2, IL-4, IL-8, IL-18, IL-1β, IFNα, TNFα) with gastritis in the age groups of schoolchildren with a family predisposal to peptic ulcer disease. To this purpose, sophagogastroduodenoscopy with biopsies of gastric mucosa was performed in the children and adolescents (7 to 17 years old) with gastroenterological complaints. A total of 143 children, who had morphologically confirmed gastritis (Sydney classification), were under study, The presence of H. pylori was also determined by morphological technique. The levels of serum cytokines were measured by ELISA technique. The inter-group differences in laboratory parameters were assessed using the Mann–Whitney test. The study has shown that in schoolchildren with gastritis and reported family predisposal for peptic ulcer, the disease is associated with increased content of IFNα in blood, both in the older age group (p = 0.001) and among younger children (p = 0.023). Meanwhile, there were no differences in IFNα content between children with hereditary burden in different age groups. In addition, in older patients with familial burden, in contrast to younger children, an increased level of IL-2 (p = 0.001) and IL-4 (p = 0.015) was noted. Among children with family predisposal to peptic ulcer and H. pylori infection, no age-dependent differences in cytokine levels were found. No differences in the cytokine contents were revealed between schoolchildren of younger and older age with highly active gastritis (grade 2-3). IL-4 contents was the only exception which was higher in older schoolchildren with grade 1 gastritis (p = 0.049). The differences in blood cytokine profile in the schoolchildren may depend on family history of peptic ulcer disease, with some age-dependent characteristics, in particular, concerning IFNα and IL-4 levels.

The phenomenon of cold airway hyperresponsiveness is rather common among patients with bronchial asthma. Possible participation of immune mechanisms in its occurence is scarcely studied. In particular, there is no information about interaction between Th17-related cytokines, and cytokines of Th2 immune response related to inflammation in asthma patients with cold-induced bronchospasm.Our objective was to evaluate the contents of IL-17A, IL-6, IL-22, IL-4 and IL-13 interleukins in asthma patients, and to specify their role in the formation of cold airway hyperresponsiveness. Spirometric indices of forced expiratory flow were measured in 43 patients with bronchial asthma. The content of interleukins in blood serum was estimated before and after bronchoprovocation test with 3-min. isocapnic hyperventilation with cold (-20 °C) air. Two groups of patients were formed with presence (group 1, n = 14) and absence (group 2, n = 29) of cold airway hyperresponsiveness, verified by the degree of forced expiratory volume reduction per 1 sec. (∆FFV_1ihca) after the cold test (-16.5 (-20.0 – -12.0)% and -2.3 (-3.5 – -0.8)%, respectively; p < 0.0001). In group 1, when compared with group 2, lower baseline values of FEV₁ (88.1±3.1% and 96.6±2.2%, p = 0.044), and forced midexpiratory flow (MEF_25-75 62.4±3.87% and 75.6±3.7%, p = 0.013) were registered. Moreover, the baseline contents of IL-17A, IL-6, IL-4 in subjects with cold airway hyperresponsiveness were significantly higher than in patients who did not respond to cold air. There was a correlation between IL-17A content in blood and severity of bronchial reaction (∆FEV_1ihca) to cold test (Rs = -0.33; p = 0.049). In asthma patients with cold airway hyperresponsiveness, high contents of IL-17A, IL-6 and IL-4 suggest a participation of both Th2 and Th1/Th17 cytokines in regulation of cold-induced bronchospasm and immune response with development of immune inflammation of “Th2 low” subtype.

Numerous observations of patients with bronchial asthma (BA) vaccinated against pneumococcal infection have revealed a significantly reduced frequency of exacerbations and hospitalisations after immunisation. Several studies have considered pneumococcal vaccines potential immunoregulators that may improve the clinical course of BA via modulation of the immune response. However, interpretation of these results has serious limitations due to heterogeneity of primary disease and differences in vaccine preparations. The aim of the present study was to perform a comparative analysis of the key cytokines levels which characterize development of distinct BA endotypes in patients following administration of conjugated pneumococcal vaccine. We have analyzed serum samples (n = 31) from patients with BA immunized by PCV13 (Prevenar 13), using ELISA technique, for Th1/Th2/Treg cytokines (IFNγ, IL-4, IL-6, IL-8, IL-10, IL-18, TNFα, and MCP-1), and total IgE level. The time points of sampling were as follows: initial terms, 6 weeks, 6 and 12 months after vaccination.
The results of the study indicate that vaccination against pneumococcal infection using a PCV13 in patients with BA was accompanied by high clinical efficacy, regardless of the disease endotype. This finding was evidenced by a decreased number of BA exacerbations in the patients, and an increased number of non-hospitalized BA patients during 1 year of observation. The clustering of patients according to their inflammatory profile enabled us to detect specific patterns of the cytokine profile. These changes included a statistically significant increase in the concentration of IFNγ in blood serum at 6 weeks after immunisation in the patients with the T2-“high” asthma endotype. Statistically significant changes were observed in patients with atopy and elevated total IgE levels in serum, who exhibited a peak increase in IFNγ concentration, also 6 weeks after vaccination. Conversely, no significant changes in cytokine levels were observed in patients with T2-“low” asthma endotype within a year after vaccination. The results of the study demonstrate that IFNγ plays a significant role in the potential adjustment of immunity in the patients with T2-«high» asthma endotype following immunisation with a pneumococcal conjugate vaccine.

During the COVID-19 pandemic, many issues remained unclear, i.e., those related to features of immune response in the disease, vaccination, and development of post-COVID syndrome, or its atypical forms. Optimal and effective vaccination strategies against respiratory viral infections in the “post-COVID era” have not been determined so far. The variety of vaccines, vaccination history, risk factors, as well as joining of COVID-19 to the spectrum of seasonal infections significantly influence the initial immunological profile of various population groups. The study group included 80 young men aged 19 years living in closed communities. Blood sampling was carried out in 2022, 9 months after the course of vaccination with the Sputnik V vaccine. The levels of pro- and anti-inflammatory cytokines (IL-1β, IL-4, IL-6, IL-8, IL-10, IL-17, IFNγ, TNFα), as well as IgM and IgG antibodies to SARS-CoV-2 were measured in venous blood sera. The results were processed by Microsoft Excel, R-Statistics, SPSS 22. The normality of distribution for quantitative data was assessed by the Kolmogorov–Smirnov test. Quantitative data are presented as Me (Q_0.25-Q_0.75); qualitative, in the form of n (%). Spearman’s correlation coefficient was used to determine statistical significance. The average level of SARS-CoV-2 IgG antibodies 9 months after vaccination exceeded the protective level by more than 3 times. Comparisons of cytokine levels and SARS-CoV-2 IgG antibodies have yielded various findings. In group 1 with SARS-CoV-2 IgG antibodies < 150 BAU/mL, the levels of pro-inflammatory cytokines (IL-1β, TNFα, IL-6, IL-8) were > 2-fold higher than in group 2 with SARS-CoV-2 IgG antibodies ranging from 150 to 500 BAU/mL. Moreover, IL-10 level was 5 times higher in group 1 compared to group 2. The level of IL-10 was significantly higher (4 times) in group 3 with SARS-CoV-2 IgG antibody levels of > 500 BAU/mL. In our opinion, there are many factors influencing the immune response, thus requiring a personalized approach to vaccination by taking into account the initial immune state. To optimize booster vaccination, the initial antibody levels should be checked, and immunological indices may be used in order to assess the necessity of re-vaccination.

The origins and intensity of human immune response to SARS-CoV-2 infection depend on many factors. The anti-infectious immune response includes both cellular and humoral components. The humoral link is mediated by effector-specific (antibodies) and non-specific factors (acute phase proteins, procalcitonin, α- and β-interferons), and regulatory factors, e.g., cytokines (interleukins (IL), γ-interferon (IFNγ)) and mediators (leukotrienes, etc.). In our work, serum levels of cytokines, i.e., IFNγ, IL-2, IL-6, IL-8, IL-10, IL-17A, IL-18 and procalcitonin (PCТ), a nonspecific effector factor of the immune system were compared at varying degrees of COVID-19 severity. Biomaterial samples (blood serum) of 91 patients aged 29-88 years (average age 53.9 years; 15 women, 76 males) were collected between May 2020 and July 2021. We divided patients into groups according to the disease severity, depending on clinical data, oxygen demand, and hospitalization data. COVID-19 patients of “mild degree” (n = 9) were subject to outpatient treatment; the patients of “moderate degree” (n = 38) were hospitalized at the infectious hospital; in the “severe degree” cases (n = 44), the patients were treated at the infectious intensive care unit. The control group of patients (n = 20) was presented by the blood donors without chronic diseases. We have identified the most important markers of the disease severity. Moreover, using ROC analysis, we have calculated the thresholds for differential diagnosis of distinct degrees of COVID-19 severity, by significantly differing criteria, as well as diagnostic efficiency of immune mediator indexes using a developed logistic regression model. We have revealed and statistically confirmed the indices that most significantly influence the COVID-19 severity, i.e., IL-6, IL-8, IL-10, IL-18, PCT, IFNγ. We found a maximum increase in IL-17A and IFNy at a mild degree, and their reduction in the moderate and maximum severe cases of COVID-19. Increase in IL-6, IL-10, PCT levels, and a decrease in IFNγ proved to be the factors of poor prognosis. We have also found some correlations between the immune mediators (IL-8, IL-17A, IL-18, IFNγ) thus allowing a deeper insight into the mechanisms of the disorders, the imbalance of immune response.

Influenza-related morbidity and mortality are disproportionately high among older population. Over 2004-2017, the highest proportion of influenza A(H3N2) virus in the WHO European Region was observed in the age group of ≥ 65 years, i.e., 10%. The efficiency of influenza vaccination is shown to be reduced among individuals over 65-70 years of age, due to age-related immune dysfunction (immunosenescence). It may be associated with an imbalance in effector memory T cells and regulatory responses. In this age group, high-dose or adjuvanted influenza vaccines are potentially more effective in terms of hospitalization days and economy than with non-adjuvanted influenza vaccines used at standard doses. The aim of our study was to investigate the ability to form a protective titer of antibodies to influenza virus among elderly persons after vaccination by the end of the COVID-19 pandemic. A total of 31 participants over 60 years of age took part in the study, who were immunized with an influenza quadrivalent inactivated subunit adjuvant vaccine by intramuscular injections. Antibodies to influenza virus strains were determined by performing a hemagglutination inhibition (HI) test one month after the vaccination. For individuals over 60 years old, one month after vaccination, a statistically significant increase in the seroprotection level (p < 0.05) was observed in relation to three strains: A/Victoria/2570/2019(H1N1) pdm09 (up to 74.2%), A/H3N2/Darwin/9/2021 (93.2%), and B/Austria/1359417/2021 (up to 74.2%). The seroprotection level to the B/Phuket/3073/13 strain was 35.5%. The geometric means of antibody titer (GMT) in older individuals before vaccination was 15.1 (log₂ 3.91±0.59) for the H1N1 strain versus 73.7 (log₂ 6.20±0.93) after vaccination; for the H3N2 strain, 52.7 (log₂ 5.72±0.97) and 147.4 (log₂ 7.20±1.22), respectively; for the B/Yamagata strain, 8.6 (log₂ 3.11±0.54) versus 24.1 (log₂ 4.59±0.79). The GMT level for the B/Victoria strain, was 10.1 (log₂ 3.33±0.38) versus 63.0 (log₂ 5.98±0.69) after vaccination. The seroconversion rate (SCR) significantly exceeded the required level of 2.00 for all strains tested (p < 0.05). For both H3N2 and B/Yamagata strains, the GMT was 2.8; for H1N1 and B/Victoria strains, 4.89 and 6.26, respectively. The seroconversion rate for H3N2 and B/Yamagata strains was 41.9%; for the H1N1 strain, 61.3%, for the B/Victoria strain it was 77.4%. The immunogenicity of each component of the influenza vaccine following a single intramuscular immunization of volunteers over 60 years old met at least one criterion of the requirements for inactivated influenza vaccines.

Methods of immunocorrection based on injection of in vitro activated autologous leukocytes are becoming widespread. Our previous studies suggest that implementation of immunotherapy with in vitro activated lymphocytes using cytokines is effective for the treatment of cancer patients, and that this therapy is well tolerated. This article discusses the use of biological drugs, i.e., recombinant interferon-gamma (IFNγ) and recombinant tumor necrosis factor-alpha-thymosin-alpha-1 (TNFα-T) for the in vitro production of human cytokine-induced killer cells (CIC). The purpose of our study was to assess the possibility of IFNγ and TNFα-T usage within protocols for in vitro production of human CIC. Isolated mononuclear blood cells from 15 donors were cultured in a CO2 incubator for 12 days and the morpho-functional characteristics of lymphocytes were studied. To assess the in vitro toxicity of IFNγ and TNFα-T towards lymphocytes, the drug concentrations have been adjusted experimentally, with respect to control of cell viability. The levels of cytokines in the supernatants at the stages of cell culture were determined by enzyme-linked immunosorbent assay (ELISA). Before and after cultivation, the presence of adhesion and granularity of the studied lymphocytes was assessed, the phenotype of cultured cells was studied by flow cytometry, and the level of viability of activated lymphocytes was assessed in a haemocytometer chamber. We carried out a comparison of methods for culturing lymphocytes under different conditions. We studied culture media containing the following supplements: only IFNγ (group 1); IFNγ, interleukin-2 (IL-2) and interleukin-15 (IL-15) (group 2); TNFα-T only (group 3); TNFα-T, IL-2 and IL-15 (group 4), and only IL-2 and IL-15 (control group). When mononuclear cells are activated in a medium containing IFNγ, IL-2 and IL-15, or in a medium containing TNFα-T, IL-2 and IL-15, we observed a general trend towards significant increase in T-cytotoxic cells (CD3 ⁺CD8⁺), NKT cells (CD3⁺CD16⁺CD56⁺), activated lymphocytes (HLA-DR⁺), activated T lymphocytes (CD3⁺HLA-DR⁺), and activation markers of all lymphocytes (CD38⁺) and on T cells ( CD3⁺CD38⁺). We assessed the possibility of using IFNγ and TNFα-T to obtain human CIK in vitro. Activated cytotoxic lymphocytes obtained by this approach may be used for fundamental research, in order to identify new patterns and morpho-functional characteristics of activated human lymphocytes, as well as for adoptive immunotherapy for cancer patients.

Recent studies suggest a role of genetic factors, i.e., polymorphisms of genes controlling the main immune components of antiviral response, and the risk of COVID-19 infection, severe course and lethal outcomes of the disease, as well as development of clinical or laboratory changes in the post-COVID period. Interleukin-2 (IL-2) plays an important role in immunopathology of COVID-19. However, there are only few data on the impact of single nucleotide polymorphisms, e.g., IL-2 330T/G (rs2069762) gene variant on molecular changes during the post-COVID period. The aim of our study was to investigate differences in systemic inflammation markers, intestinal permeability and vascular regulation in patients with post-COVID syndrome, and single nucleotide polymorphisms (SNPs) of the IL-2 330T/G gene. Fiftyfour patients (28 females (51.85%) and 26 males (48.15%), mean age 45.6±6.14 years) who suffered with COVID-19 were included into the study. The patients were tested for IL-2 T-330G polymorphism by allelespecific polymerase chain reaction (PCR) with electrophoretic detection of products. PCR was performed using the IL-2 T-330G kit (LLC "Litech", Russia). The contents of C-reactive protein (CRP, mg/L), lipopolysaccharide-binding protein (LBP, ng/mL), tissue plasminogen activator (tPA, ng/mL), zonulin (ng/ mL), endothelin-1 (pg/mL), and angiotensin-2 (pg/mL) in blood plasma were determined by ELISA test manufactured by Cloud Clone Corp. (Wuhan, Hubei, China). The patients harboring a homozygous GG variant of T-330G IL-2 gene polymorphism showed significantly lower CRP levels than in the heterozygous TG group (p = 0.013), and TT homozygous group (p = 0.039). The tPA levels were significantly higher in the GG homozygote group compared to TT homozygotes (p = 0.017). The highest zonulin values were recorded in the TG heterozygote group, compared to TT homozygotes (p = 0.013). The highest angiotensin-2 values were found in the homozygous TT group (p < 0.05). No significant variations of LBP and endothelin-1 were registered between the studied groups. The T-330G polymorphism of IL-2 gene is, therefore, associated with some molecular changes in the post-COVID period, which may potentially influence both clinical manifestations of post-COVID syndrome, and its long-term consequences in future. Further in-depth studies of T-330G effects upon activity of the IL-2 gene and related molecular events is necessary in order to understand the clinical aspects of post-COVID syndrome. 

The relation between polymorphic variants of the HLA-G gene and various pregnancy complications (failure after in vitro fertilization, preeclampsia, spontaneous abortions) is discussed. However, research results vary among women in different populations. It remains unknown whether maternal HLA-G alleles can control congenital malformations (CMs) in the fetus. We studied the role of HLA-G gene polymorphism in the formation of pathology in in maternal-fetal interface using the example of recurrent miscarriage (RM) and CMs in the fetus. We studied 461 women with reproductive pathology and 407 healthy and fertile women having 1-2 children and with no history of pathological pregnancy (control group). The RM group included 151 women with miscarriages before 20 weeks of pregnancy (min = 2; max = 6). The CM group consisted of 310 women with congenital malformations of the fetus. The diagnosis of CM type was carried out according to the International Classification of Diseases, Injuries and Conditions Affecting the Health, Tenth Revision (Q00-Q99). All women provided written informed consent to participate in the study. The rs41551813, rs12722477 and rs41557518 loci of the HLA-G gene were typed by asymmetric real-time PCR. HLA-G 14 bp Ins/Del polymorphism (rs66554220) was determined using electrophoretic separation of amplification products. Polymorphic loci rs41557518 and rs66554220 of HLA-G were linked disequilibrium (D’ = 0.808 (r = 0.017), χ² = 14.67, d(f) = 3, p = 0.002). Found only highly significant the association of the 110Ile HLA-G allele with the risk of RM in women after statistical correction (OR = 3.03 (1.97-4.64), p_cor < 0.0006). Found no statistically significant associations of polymorphic loci rs41551813, rs41557518 and rs66554220 of the HLA-G gene with RM in women. Found no associations of maternal HLA-G polymorphic loci with the risk of CMs in the fetus. It seems the role of individual the HLA-G polymorphic loci in the formation of CMs in the fetus may be minimal. The association of the 110Ile allele (HLA-G*01:04) with RM in women is likely due to its recessive effects on local inflammation.

YB-1 is a multifunctional protein, being a transcription factor involved in the regulation of numerous cellular processes. YB-1 is an important factor in molecular cascades that regulate the response to the pathogen invasion, inflammatory activity, as well as efficient curation and healing. We suggest that YB-1 may also play an important role in pulmonary tuberculosis. However, the YBX1 transcriptional activity and the role of the YB-1 protein in pathogenesis of this disease have not yet been determined. The aim of our study was to identify the most significant correlations between expression rates of YB-1 gene with expression of some key cytokine genes involved in the regulation of tuberculous inflammation (IL-6, IL-10, IFNγ, TGF-β, TNFα, IL-1β), hypoxia factor-1 (HIF1a) gene, and P-gp protein gene ABCB1 in the patients with pulmonary tuberculosis. Gene expression was determined by quantitative PCR in the samples obtained at surgery from 35 patients. Correlation and cluster analysis were performed based on the PCR results. A positive correlation was found between the expression of YBX1, TGFB1, and ABCB1 genes. Correlation between the expression of YBX1 and ABCB1 genes were moderate, whereas ABCB1 gene expression exhibited a strong positive correlation with HIF1A and IL6 genes. The strongest correlation was found between YBX1 and TGFB1 gene expression (r = 0.62). There is no correlation found between YBX1 and the genes encoding other cytokines. TGFB1 showed a moderate correlation with TNF (r = 0.56). The relationship of YBX1 with TGFB1 was confirmed by cluster analysis, thus demonstrating a single cluster of YBX1, TGFB1, TNF. We assume that the YBX1, TGFB1, TNF gene cluster forms a regulatory system that plays an important role in development of tuberculous inflammation. Our work expands the knowledge on the molecular genetic features in tuberculoma, a clinical form of pulmonary tuberculosis. We suggest that the YB-1 protein can potentially have different functions: (1) being a participant in tuberculous inflammation via the cytokine expression; (2) modulating the P-gp activity and changing the pharmacokinetics of anti-tuberculosis drugs, thus requiring future studies.

The immunomodulatory properties of probiotics largely depend on the metabolites secreted into the culture medium, which is studied as a cell-free supernatant and referred to as postbiotics. The aim of the present work was to conduct screening of cytokine profiles for intestinal strains of Bifidobacteria and Lactobacillus by testing it with mononuclear cells from human peripheral blood, and comparing it with cytokine profile of typical industrial bacterial strains, in order to select promising strains with anti-inflammatory properties as potential pro-/postbiotics. The cytokine profile of probiotic and intestinal strains of Bifidobacteria and Lactobacillus isolated from the large intestine of healthy people was determined by the biological effects of cell-free supernatants on the production of pro- (IFNγ, TNFα, IL-17, IL-8, IL-6) and anti-inflammatory (IL-10, IL-1ra) cytokines in the in vitro model of peripheral mononuclear cells isolated from human blood. We have established three types of effects on the cytokine profile: type 1 was characterized by a predominant increase in IL-10 production, and a decrease in TNFα, IL-17, IL-6; type 2, produced a decrease, mainly at the level of pro-inflammatory cytokines; type 3 caused a decreased secretion of both pro- and anti-inflammatory cytokines. Among type 1 and type 2 cultures, the Bifidobacterium bifidum ICIS-202 and Bifidobacterium bifidum ICIS-504 strains had high anti-inflammatory potential, capable of both suppressing the secretion of pro-inflammatory cytokines, and enhancing the production of anti-inflammatory cytokines. The supernatant of L. ruminis ICIS-540 strain showed a promising effect, i.e., it repeatedly reduced the level of early proinflammatory TNFα cytokine. The anti-inflammatory activity of these strains was not inferior, but, in relation to individual cytokines (IL-10, TNFα, IL-6), was superior to the known probiotic bacterial cultures. The in vitro testing of metabolic products in bacterial supernatants enabled us to select promising strains of L. ruminis ICIS-540, B. bifidum ICIS-504 and B. bifidum ICIS-202 which may be suitable for implementation of bacterial preparations with anti-inflammatory activity.

Decidual NK cells exhibit distinct phenotypic and functional characteristics as compared to peripheral NK cells. However, the mechanisms underlying development of these unique properties remain poorly understood. The cells in microenvironment are known to exert both direct and indirect influence on NK cells within uterus, modulating their level of “aggressiveness” towards fetal tissues, including trophoblasts. Cytokine release presents a remote regulatory tool for the NK cells. Trophoblasts produce cytokines like as other components of the microenvironment. These cytokines bind the receptors on surface of target cells thud changing the behavior of NK cells. As a result, NK cells may release the own cytokines, which, in turn, influence the behavior of other cells. As mentioned above, there is a lack of data on causes and mechanisms behind the changes in characteristics of NK cells in uterus. Nevertheless, this data can lay the foundation for designing a more accurate cellular model of interactions between fetal cells and maternal immune system. Moreover, it may serve as a basis for developing diagnostic tools for reproductive issues.
The aim of our study was to investigate changes in cytokine profile of NK cells, in particular, their production of TNFα, TGF-β, IFNγ, RANTES, IL-10, and VEGF under the influence of cytokines associated with pregnancy, i.e., TNFα, IFNγ, TGF-β1, IL-15, IL-18, or IL-10. The levels of these cytokines in the culture media conditioned by NK cells were measured using flow cytometry. TGF-β1, produced by trophoblasts was found to have the ability of regulating cytokine secretion by NK cells. The levels of IFNγ, IL-10, and RANTES in the media derived from NK cell culture have been decreased under its influence.
On the basis of these findings, one may propose the existence of a regulatory system that controls activity of NK cells via the cytokine network. These data suggest a potential for using TGF-β1 to model in vitro interactions between NK cells and trophoblasts.

Chronic diffuse liver diseases are among urgent problems in the modern world. Exposure to tobacco smoke was used as a model to reproduce liver pathology. The purpose of the study was to evaluate the level of cytokines, i.e., IL-6, IL-18, IL-1α, IFNγ, TNFα, TGF-β1 in an animal model of tobacco intoxication, with administration of Heptor substance for 7, 14, 21 days. The studies were carried out on male rats, divided into intact and 3 experimental groups of 10 rats each. The animals were exposed to tobacco smoke for one, two and four months. For the experiments, a chamber was used with a volume of 0.3 m³ , in which the cigarette smoke was produced. After four months of exposure (3^rd group), Heptor was administered per os at a dose of 120 mg/kg with distilled water for 7, 14, 21 days. After withdrawal from experiment, the liver specimens were sampled, in order to study the level of cytokines. Liver sections were also prepared and stained with Hematoxylin/Eosin. In group 1, inflammatory cell infiltration and degenerative changes were detected in the liver; fibrosis was absent. In the 2^nd group, necrobiotic foci were noted in the liver along with hemorrhages and dystrophic changes; fibrosis was absent. In the 3^rd group, destructive changes, areas of necrosis in the liver, porto-portal and porto-central septa were observed. The level of TNFα was 1.6 times increased in the cytoplasmic fraction of liver tissue in the 1^st group compared to the intact group; in the 2^nd group it was 1.7 times higher; in the 3^rd group it increased by 1.9 times. The concentration of IL-6 was the highest in the group 3. At these terms, TGF-β1 was significantly increased, with destructive changes and liver fibrosis being detected by histology. The use of the drug Heptor for 21 days in animals of the third group contributed to decreased levels of pro-inflammatory cytokines (IL-18, IL-6, IFNγ) in liver tissue, along with reduced TNFα and TGF-β. As a result of their influence, a decrease in the inflammatory response, destructive changes and lower degree of fibrosis was noted, as well as gradual restoration of the liver structures was developed.

Recently, flow cytometry (FC) has been used both for scientific purposes and in routine laboratory practice. The range of tasks solved by the FC technique is constantly expanding, and flow cytometry is currently used not only for the diagnosis of hemoblastoses and evaluation of therapeutic efficiency, but also is a promising method for analyzing functions of various cell populations, including blood neutrophils, which play a role in pathogenesis of different immunodeficiency conditions accomplished by recurrent bacterial infections. This work concerns implementation of an FC-based method for determining phagocytic activity of neutrophil granulocytes into laboratory practice. In the course of this study, this test was compared with a traditional technique used in laboratories, i.e., simple visual assessment of phagocytosis reaction of baker’s yeast in suspension. Determining the phagocytic activity of neutrophils by means of flow cytofluorimetry proved to be more reproducible and easily controlled when compared to simple visual assessment technique. We have found that the results of studying the phagocytosis parameters measured by FC-based technique were not influenced by gender and age of the patients. Therefore, uniform ranges for the reference intervals were calculated for different categories of patients. Clinical testing of the FC-based method for determining phagocytic activity of neutrophils was performed in a group of patients suffering from recurrent bacterial infections. Assessment of phagocytic activity by FC approach enabled us to identify a patient with phagocytosis deficiency. Meanwhile, the method of simple visual phagocytosis with a suspension of baker’s yeast showed inferior sensitivity in this case, and the obtained indices of neutrophils’ phagocytic activity did not exceed the reference intervals. Thus, the FC-based method of determining phagocytic activity of neutrophils using FC approach is preferable when used in routine laboratory practice as compared with the method of simple visual assessment of baker’s yeast phagocytosis.