Extracellular vesicles (EV) are currently considered potential biomarkers of diseases, as well as “biological constructs” for targeted drug delivery. Extracellular vesicles represent a heterogeneous population of membrane vesicles formed by various cell populations, including immune cells. At present time, EV are divided into exosomes, microvesicles, and apoptotic bodies, depending on their size and formation manner. EV have been found in various human biological fluids. Therefore, the possibility of their usage as diagnostic biomarkers is under discussion. The vesicles have a diverse internal composition and express a wide repertoire of receptors on their surface, thus allowing them to participate in different intercellular communications by transferring to the cells various molecules, including genetic material. E.g., microRNAs transmitted via extracellular vesicles are evolutionarily conserved non-coding RNA molecules 18-25 nucleotides long. Their main function is to regulate gene expression at the post-transcriptional level. MicroRNAs are synthesized by different cell types. However, some microRNAs are found ubiquitously, whereas others are present only in certain types of tissues. MicroRNAs are found both inside, and outside the cells (extracellular or circulating microRNAs). The microRNAs are resistant to RNases and stable in the extracellular environment, due to their secretion as protein complexes, or as part of extracellular vesicles. The variable microRNA profile in extracellular vesicles depends on the physiological conditions and presence of various pathological disorders. Multiple studies show that microRNAs can determine the functional activity of extracellular vesicles, e.g., therapeutic usage of microRNAs carried by EV as well as diagnostic applications in various pathologies. This review considers distinct populations of extracellular vesicles and their main properties, describes the characteristics of intra- and extracellular (circulating) microRNAs, mechanisms of their biosynthesis, and techniques for detection and assessing contents of microRNAs. The review describes microRNAs as a component of exosomes and microvesicles formed by various cells, including cells of the immune system in the course of physiological and pathological processes, with respect to functions of these microRNAs as well as their diagnostic and therapeutic potential.

Chronic rhinosinusitis (CRS) is a disease caused by inflammation of the paranasal sinuses and its mucous membrane lasting for more than 4 weeks continuously. The aim of our study was to examine the main pathophysiological features of chronic IgE-mediated rhinosinusitis of bacterial etiology according to publications in the Russian Federation and in the world. A search was made through English- and Russian-language literature sources using the following databases: PubMed, MedLine, Web of Science, Russian Science Citation Index, Springer, Scopus, Scientific Research, Google Scholar, Crossref, eLibrary. The epidemiological features of CRS in the Russian Federation, bacterial pathogens and pathophysiological characteristics of CRS were analyzed. A 2-fold increase in the prevalence of CRS was registered over the past 20 years. Prevalence of the disease increases at longer age ranges. Chronic rhinosinusitis ranks first among all chronic diseases in the field of otorhinolaryngology. Allergic rhinitis, asthma, bronchiectasia, immunodeficiencies, cystic fibrosis, primary ciliary dyskinesia and autoimmune diseases are associated with CRS. The most common bacterial pathogens are S. aureus, Staphylococcus epidermidis and Propionibacterium acnes, Prevotella, Streptococcus and Veillonella, and some Gram-negative bacteria, e.g., Pseudomonas aeruginosa (P. aeruginosa), Proteus mirabilis and Klebsiella pneumoniae. Staphylococcus aureus (S. aureus) is involved in pathogenesis of nasal polyps. The colonizing bacteria may contribute to pathogenesis of CRS through the formation of biofilms. Alterations in the sino-nasal microbiome may also contribute to the development of CRS. An association of the CRS and CFTR gene mutations plays a significant role in the pathogenesis of chronic rhinosinusitis. An “immune barrier hypothesis” has been proposed as potential mechanism of CRS. Reduced expression of SPINK5, impaired STAT3 signaling, and T2R38 bitter taste receptor polymorphism have been identified in the pathogenesis of CRS. The T2R38 gene stimulates epithelial cells to produce nitrous oxide with a bactericidal effect, promotes mucociliary elimination of pathogens and prevention of upper respiratory tract infections, the polymorphism of this gene predisposes patients to gram-negative infectious diseases, and therefore is a risk factor for the development of CRS. In addition, antibody deficiency is the most common primary immunodeficiency associated with CRS.

Hence, the pathogenesis of chronic IgE-mediated rhinosinusitis of bacterial etiology is associated with defects in innate immunity and mucociliary clearance, influence of the sinonasal microbiome, allergies, and genetic factors. A comprehensive assessment of these factors is necessary for the development of new preventive and therapeutic options for the correction of CRS.

Recent expansion of fundamental knowledge on the physiology of lactation, and breast milk exosomes, stem cell biology, mother-child interactions from prenatal period to postnatal development requires a progressive, dynamic view from the scientific community and practicing physicians when analyzing known, generally accepted clinical phenomena and patterns (development of the immune system of infants and young children, natural and artificial feeding, features of postnatal development and growth of organs and tissues in children born prematurely). The components of the mother-breast-milk-infant triad are closely related to each other and influence developmental trajectory of the infant. According to modern concepts, breast milk of a nursing woman is a “living, metabolic / endocrine signaling system”, which may be considered an “immune organ” significant for postnatal growth and body programming of a premature baby. A valuable phenomenon of early postnatal development is actively discussed in the special literature, i,e., “microchimerism” caused by breastfeeding which, according to modern concepts, may play a key role in development of immune system and the whole body. Absence of protective (immunomodulatory and regenerative) effects of breast milk from a nursing woman on the spontaneous, uncorrectable impact of adverse factors of prematurity is likely predispose for remodeling and dysfunction of heart in prematurely born children, and, at longer range, in adults. The young children born prematurely show a unique cardiac phenotype characterized by reduced biventricular volume, relatively lower systolic and diastolic function, disproportionate muscle mass gain, clinically manifesting by increased risk of cardiovascular disease, hypertension, and decreased exercise tolerance. Hence, the premature birth may be considered a chronic disease state. Therefore, the natural feeding which provides a natural evolutionarily protective mechanism for the child’s heart should be attributed to the fundamental factors that play a vital role in prevention of cardiovascular diseases in prematurely born children and at later life periods.

The purpose of the study was to evaluate the severity of changes in the values of markers-candidates for the differential diagnosis of anemia of chronic diseases in patients with type 1 and type 2 diabetes mellitus. The total number of leukocytes, erythrocyte sedimentation rate, content of C-reactive protein, TNFa, ferritin and hepcidin were evaluated. 50 people with type 1 diabetes mellitus and 81 people with type 2 diabetes mellitus were examined. The diagnosis of anemia was established on the basis of data on the level of hemoglobin, the content of erythrocytes in the blood, ferritin and serum iron. Next, the type of anemic syndrome was determined. The patients were divided into groups: 14 patients with diabetes mellitus and anemia of chronic diseases, 15 people with diabetes mellitus and iron deficiency anemia, 38 patients with diabetes with latent iron deficiency and 64 patients with diabetes mellitus without anemia. The comparison group consisted of 17 healthy volunteers. It was shown that in the general sample of patients with diabetes mellitus anemia of chronic diseases was distinguished only by the erythrocyte sedimentation rate, which was higher than in iron deficiency anemia, latent iron deficiency and in patients without anemia. The severity of inflammation in diabetic patients was analyzed depending on its type. The concentration of hepcidin in the blood of diabetic patients, regardless of type, exceeded its content in the blood of healthy individuals. Elevated serum concentrations of TNFα were characteristic of inflammation in type 1 diabetes mellitus. Diabetes mellitus type 2 was characterized by an increase in: erythrocyte sedimentation rate - relatively healthy individuals; concentrations of C-reactive protein - in comparison with healthy volunteers and patients with type 1 diabetes mellitus; ferritin levels compared with patients with type 1 diabetes mellitus. Taking into account the type of diabetes and the type of iron metabolism disorder, it was found that in type 1 and type 2 diabetes mellitus, only the erythrocyte sedimentation rate in patients with anemia of chronic diseases was significantly higher than in patients with iron deficiency anemia and without anemia. The article discusses the reasons for the difficulties in using inflammatory markers (ferritin and hepcidin) as parameters for verifying anemia of chronic diseases in patients with diabetes mellitus. It is pointed out that it is necessary to take into account the differences in the mechanisms of inflammation development in type 1 or type 2 diabetes mellitus when trying to use cytokines and C-reactive protein as additional diagnostic markers in practice. The rationale is given for the prospects of determining the erythrocyte sedimentation rate, with the recommendation of a certain threshold value, for the detection of anemia of chronic diseases in patients with type 1 and type 2 diabetes mellitus.

The world is experiencing a rapid increase in the prevalence of allergic and autoimmune diseases. It is known that allergic inflammation is most often systemic, involving various organs and systems in the pathological process, such as the skin, respiratory and gastrointestinal tract with the development of dermatorespiratory, dermato-intestinal and other manifestations. The study of the features of the cytokine profile in oral fluid (saliva) deserves special attention, since these characteristics reflect not only local, but also systemic disorders. Of particular relevance is the study of local cytokine regulation of intercellular interactions in food allergies. Our objective was to study the concentration of IL-4, IL-10, IFNγ, secretory IgA in salivary fluid, the concentrations of total immunoglobulin E and eosinophilic cationic protein in blood serum of the patients with atopic dermatitis and psoriasis with concomitant food allergies.
The study included patients with atopic dermatitis (AD, group 1, n = 20), psoriasis with concomitant food allergy (PS, group 2, n = 27), psoriasis without concomitant allergies (PS, comparison group 3, n = 23). Quantitative assessment of the cytokine concentrations (IL-4, IL-10, IFNγ, sIgA) in salivary fluid was carried out by enzyme-linked immunosorbent assay. Concentrations of total immunoglobulin E and eosinophilic cationic protein in blood serum were determined by indirect immunofluorescence. The obtained results were processed using the Statistica 8.0 applied software.
In groups of patients with atopic dermatitis (Group 1) and psoriasis with concomitant food allergy (Group 2), we have noted a statistically significant increase of salivary IL-4 and IL-10, as well as of total immunoglobulin E concentrations in blood serum as compared with a group of patients with psoriasis without concomitant allergies (group 3), and with control group. When studying concentrations of IFNγ in saliva, no statistically significant intergroup differences were found. The concentration of sIgA in saliva was significantly higher in the groups of patients with atopic dermatitis and psoriasis accompanied by food allergies in comparison with control group and the group of psoriatic patients without food allergies (group 3).
The cytokine profile of saliva is characterized by unidirectional changes in food allergy. Skin seems to be the shock organ in this condition, regardless of nosological form of the disease (atopic dermatitis or psoriasis). Salivary fluid is an easily accessible material when assessing the state of mucosal immunity in food allergies.

Increased incidence of alopecia has been noted in children at the present time. Participation of autoimmune (immunopathological) mechanisms in pathogenesis of this disease necessitates additional study of immune status and characteristics of comorbid pathologies. The aim of our study was to specify the features of immune status and comorbidities in children with alopecia areata. The observation group consisted of children with various types of alopecia areata (n = 57), a comparison group included children without clinical manifestations of alopecia or a history of alopecia (n = 157). We performed a comparative evaluation of major lymphocyte subpopulations (CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺, CD3⁺CD19⁺), interleukins (IL-4, IL-6), immunoglobulins (IgA, IgM, IgG), parameters of phagocytic activity (absolute phagocytosis, percentage of phagocytosis, phagocytic number and phagocytic index) and IgE to house dust and cat hair. Analysis of comorbidities was also performed. Statistical processing was carried out with Jamovi software. We have found that the focal clinical form of alopecia prevailed over the subtotal and total forms by 1.8 times (p = 0.033) in the observation group rather than in comparison group, with common variable immunodeficiency being more often (1.4-fold), chronic tonsillitis (3.9-fold), allergic rhinitis (3.9-fold) and autoimmune thyroiditis, which was absent in the comparison group. Among the children with alopecia, disturbances of the T-cell link were revealed, i.e., a higher median levels of the relative and absolute numbers of CD3⁺CD4⁺ cells (p = 0.001 to 0.003), larger proportions of elevated values for IgA (4.1-fold), IgM (7.3-fold), IgG (13.2-fold) with p-levels of 0.0001 to 0.0008; increased IL-4 (8.1-fold) and IL-6 (4.6-fold), with p = 0.002-0.004, along with medium and relatively strong correlations with alopecia. In children with alopecia, we have determined a 3.3-fold proportion of reduced values of absolute phagocytosis and 3.7-fold reduced percentage of phagocytosis (p = 0.0012 to 0.028), with an sufficient correlation (weak to moderate strength) with alopecia, as well as lower values of the median phagocytic index and phagocytic number in the observation group (p < 0.001) associated with a average-strength correlation. Hence, an imbalance of the immune system components was revealed in children with alopecia which manifested with signs of immune hyperfunction, characteristic, e.g., of autoimmune and allergic processes, accompanied by more frequent registration of autoimmune thyroiditis and allergic rhinitis. Moreover, distinct signs of immune deficiency, are found, characterized by a decrease in phagocytic activity and higher incidence of common variable immunodeficiency and chronic tonsillitis.

The aim of our study was to evaluate the features of HLA-G and HLA-DR expression on lymphocytes of women and their children with congenital heart defects (CHD) under the influence of allogeneic and autologous blood sera.

38 women and their children with sporadic septal congenital heart defects (main group) were examined. The comparison groups included 21 women and their children without congenital heart disease (comparison group 1), as well as 17 apparently healthy men (comparison group 2). A total of 115 individuals were examined. The cross-match studies were carried out using a CytoFlex flow cytometer (Beckman Coulter, USA). The effects of autologous and allogeneic blood sera on HLA-G and HLA-DR expression on lymphocytes were evaluated. Statistical processing of the obtained results was carried out using Statistica for WINDOWS software packages from StatSoftInc. Version 10.0 and MedCalc 17.5.3. by the rules of variation statistics.

The expression of HLA-G and HLA-DR molecules on the lymphocytes did not significantly change under the influence of autologous serum from men and women of children with CHD. At the same time, in women with more than two births of apparently healthy children, autologous serum significantly suppressed expression of HLA-G and HLA-DR on their lymphocytes. In particular, a pronounced and significant suppression was noted with autologous serum for HLA-DR molecules on CD3-positive lymphocytes. One may suggest that inflammation in the mother-embryo system is limited by this mechanism. Other significant differences concerned the effect of autologous and allogeneic (maternal) sera on the expression of HLA-G and HLA-DR molecules on the children’s lymphocytes. We have shown that in the group of children with septal CHD, autologous and allogeneic sera did not suppress the expression of HLA-G and HLA-DR on lymphocytes. At the same time, in the group of apparently healthy children, autologous and allogeneic (maternal) sera suppressed the expression of HLA-G and HLA-DR on lymphocytes. Moreover, the suppressive effect upon expression of both HLA-G and HLA-DR was significantly higher in allogeneic (maternal) sera than in autologous serum (p < 0.01). This effect seems to be determined by the presence of autoand alloimmune antibodies to HLA-G and HLA-DR molecules in blood serum of multiparous women.

The suppressor activity of female sera against allogeneic (embryo / fetus / child) and autologous (intrinsic) HLA-G and HLA-DR antigenic molecules may determine a protective effect related to development of septal congenital heart defects in offspring.

Today, the diagnosis and treatment of severe infectious and inflammatory diseases in newborns, e.g., congenital pneumonia (CP) and neonatal sepsis (NS), present difficult problems. Searching sensitive and specific severity markers of bacterial inflammatory process as well as early and effective treatment are crucial for the outcome and prognosis of these life-threatening diseases. The aim of our study was to assess the effects of intravenous immunoglobulin (IVIG) injections on the negatively transformed subpopulations of neutrophilic granulocytes (NG) СD64^-CD16⁺СD32⁺СD11b⁺, СD64⁺CD16⁺СD32⁺СD11b⁺ and evaluation of their functional activity in newborns with CP and NS. We have observed 38 full-term newborn patients. Group 1 included 19 infants with CP, including 11 children who received conventional therapy and IVIG (group 1.1), and 8 children treated at conventional protocols (group 1.2). Group 2 included 19 children with NS, including 12 children who underwent conventional therapy and IVIG treatment (group 2.1), and 7 children who were subject to conventional therapy (group 2.2). The comparison group consisted of 22 healthy full-term newborns. Testing of NG population included the following parameters: counting the numbers of NG subpopulations which simultaneously expressed CD11b CD64, CD32, CD16, as well as their phenotypic patterns, with regard of the receptor expression density (MFI) using flow cytometric techniques. Moreover, we determined phagocytic and microbicidal activity of the granulocytes. We have revealed negative transformation of СD64^-CD16⁺СD32⁺СD11b⁺ and СD64⁺CD16⁺СD32⁺СD11b⁺ subpopulations of neutrophilic granulocytes in newborns with CP and NS, The diagnostic significance of increased СD64⁺CD16⁺СD32⁺СD11b⁺NG subpopulation was more pronounced with increasing severity of bacterial infection and inflammatory process, i.e., 18.7-fold in CP, 52.3-fold in NS, along with predominant decrease in expression of appropriate membrane receptors. These phenotypic changes were associated with impaired phagocytic and killing activity of NG. The effect of IVIG on the impaired mechanisms of antibacterial immunity is associated not only with alleviation of IgG deficiency, but also with positive remodeling of negatively transformed subpopulations of СD64^-CD16⁺СD32⁺СD11b⁺NG and СD64⁺CD16⁺СD32⁺СD11b⁺NG, improved effector functions of NG, especially in cases of CP. Thus, following IVIG treatment, a reduced number of СD64^-CD16⁺СD32⁺СD11b⁺NG subpopulations was fully recovered in CP, while it increased 1.5 times in NS, and the content of diagnostically significant СD64⁺CD16⁺СD32⁺СD11b⁺NG subpopulation showed a significantly decrease, both in CP (2-fold) and in NS (2.6-fold). However, this index remained higher than the content of this subpopulation in healthy newborns. At the same time, we have noted the restorative or modulatory effects by changing density of trigger molecules in NG subpopulations. Limitation of the negative NG transformation in their functionally significant subpopulations in newborns with CAP and NS was accompanied by positive clinical effects, i.e., optimization of antibiotic therapy, reduced duration of treatment, and improved mortality rates.

Hematopoietic stem cell transplantation is used to treat hemoblastoses and some other diseases. Depending on the diagnosis, autologous cells of the patient or allogeneic cells obtained from a healthy related or unrelated donor are used. A sufficient count of harvested hematopoietic progenitor cells is necessary for successful transplantation. Currently, stimulation of their egress from the bone marrow into the peripheral blood by a granulocyte colony-stimulating factor, followed by collection by leukapheresis, is widely used for their preparation. Hematopoietic stem cells leave the bone marrow niche formed by the stromal microenvironment when their holding bonds are interrupted. Mesenchymal stromal cells regulate the release of hematopoietic precursors by producing various cytokines and other biologically active substances. Therefore, the study of the functional properties of bone marrow mesenchymal cells can help in solving the problem of “poor” mobilizations that occur in patients with multiple myeloma.

The aim of the study was to estimate the effectiveness of mobilization of hematopoietic stem cells depending on the cytokine-producing ability of mesenchymal stromal cells in the bone marrow of donors and patients with multiple myeloma.

The yields of hematopoietic progenitors were studied in 10 donors (median age 27 years) and 18 patients with multiple myeloma (median age 57 years). In donors, the release of hematopoietic stem cells into the peripheral blood was stimulated by subcutaneous administration of G-CSF at a dose of 10 μg/kg/day. Patients with multiple myeloma received vinorelbine at a dose of 35 mg/m² of body surface, followed by administration of G-CSF (10 μg/kg/day subcutaneously). A culture of mesenchymal stromal cells was obtained from bone marrow taken prior to mobilization and the content of interleukins IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-α and IFN-γ in supernatants was studied by enzyme-linked immunosorbent assay using kits of reagents produced by Vector-Best (Novosibirsk).

Patients with multiple myeloma were characterized by high secretion of the pro-inflammatory cytokine IL-2 (4.70 pg/ml vs 3.55 pg/ml in healthy individuals, p=0.003) and low IFN-γ (0.41 pg/ml vs 2.14 pg/ml in healthy, p<0.001), but no relationship was found between these cytokines and the hematopoietic stem cells yield. The present study showed that in patients with ineffective harvesting of hematopoietic precursors, secretion of IL-8 by stromal cells was low (308.08 pg/ml vs 561.29 pg/ml in healthy individuals, p=0.04).

The obtained results are consistent with the literature data on the important role of IL-8 in the mobilization of hematopoietic stem cells, which allows to consider IL-8 as an informative marker for predicting the insufficient yields of hematopoietic precursors.

Experimental medicine provides the scientific community with a plethora of information on therapeutic efficacy of probiotic strains. However, from the point of view of evidence-based medicine, the list of disorders controlled by probiotics is limited to antibiotic-associated diarrhea in adults and children, Clostridium difficile-associated diarrhea, acute infectious diarrhea in children and adults, eradication therapy, ulcerative colitis and irritable bowel syndrome. Recently, these indications are also amended by well-validated clinical guidelines for the usage of probiotic preparations, in order to modulate immunity. Given the permeability of gastrointestinal and immune system barriers for pathogenic and opportunistic microbiota, it seems logical to assume the effectiveness of probiotics as potential symbiotic regulators of nervous and cardiovascular systems. It should also be taken into account that metabolic disorders, e.g., obesity, with a low-intensity inflammatory response and characteristic cytokine pattern, are acquired as a gain of human civilization. In this regard, we propose a scientific hypothesis about the effectiveness of probiotic microbial strains in increasing myocardial resistance to ischemic-reperfusion injury, due to their ability to block individual links of the cytokine cascade during the development of inflammatory response, for its subsequent translation into clinical practice.

The development and validation of a new experimental model of systemic inflammatory response syndrome (SIRS) in male Wistar rats, including obesity, acute inflammatory process of the colon, and antibiotic-induced dysbiosis, became basic to the study of efficacy of probiotic drugs in terms of myocardial resistance to ischemicreperfusion injury (IRI). Rats with SIRS showed a significantly increased size of the infarction area (+28%) upon experiments with isolated perfused heart under global ischemia-reperfusion conditions. Significant changes in the leukocyte formula and immunological parameters associated with SIRS were corrected by introduction of a mixture of probiotic strains L. acidophilus (LA-5) and B. animalis subsp. lactis (BB-12), and the isolated strain L. delbrueckii TS1-06. In both groups with probiotic correction, there was a decrease in the infarction area compared to the SIRS group. General and specific changes in IL-2, transforming growth factor-b (TGF-b) and tumor necrosis factor-a (TNFa) were noted. The reduction of myocardial infarction by probiotics may be related to the blocking of first-order cytokines, which leads to a «break» of proinflammatory cascade. A need for in-depth study of cardioprotective mechanisms mediated by probiotics was confirmed due to their potential usage as a symbiotic alternative to biological drugs which block the main pro-inflammatory cytokines.

Tyrosine protein phosphatase (common leukocyte antigen CD45) regulates FcᵧR-mediated cell signaling and secretory function of neutrophilic granulocytes (NG) when interacting with antigen-antibody immune complexes. The aim of the work is to study changes in the expression of CD45 on the surface of human granulocytes in ex vivo modeling of bacteremia by live cells of the plague microbe vaccine strain Yersinia pestis EV NIIEG and to evaluate the priming effect of the live plague vaccine (LPV) in vivo in terms of this parameter. The expression density of CD45 on NG was determined by flow cytometry in conventional units of fluorescence intensity (MFI) after staining the cells with the CD45-FITC labeled mouse antibody reagent (Backman Coulter, USA) during immunophenotyping of blood leukocytes according to the Lyse/No Wash protocol. In donors not previously vaccinated against plague (group 1), the value of the indicator was assessed before and 30 min, 2 h, 6 h after the addition of Y pestis EV cells to whole blood at a dose of 10⁸ mc/ml, as well as 1 month and 6 months after the primary anti-plague vaccination. In individuals who had previously been repeatedly vaccinated with LPV in the territory of the natural plague focus (group 2), CD45 expression on blood granulocytes was determined one year after the last annual vaccination, and then 1 month and 6 months after revaccination. Getting into human blood, living cells of the vaccine strain Y pestis EV of the plague microbe induced a change in the NC phenotype already after 30 minutes, associated with a 3.5-fold increase in the surface expression of CD45, which remained at an elevated level for 6 hours. The studied indicator depended ex vivo on the degree of resistance of plague microbes to phagocytosis and killing of NG. Plague vaccination had a similar stimulating effect on human peripheral blood NG in vivo. Under the influence of HPV, CD45 expression increased on blood NG in groups 1 and 2 one month after vaccination, and the changes persisted in volunteers for 6 months. The experimental data obtained in the work may reflect the result of NG priming with lipopolysaccharide and other Y. pestis antigens. The registered functional activation of NG by expression of tyrosine protein phosphatase probably indicates the formation of “immune memory” at the level of innate immunity cells under the influence of LPV, the functioning of which explains the development of a faster and more intense antigen-specific immune response to repeated introduction of the plague vaccine into the body. 

Polyphenols exert a wide range of biological effects, including immunomodulatory action. Studying the effects of flavonoids on phagocytic activity of specialized phagocytic cells seems to be a rather promising direction for their further usage as pharmacological (therapeutic) agents. Quercetin and luteolin are the most commonly studied flavonoid substances with pleiotropic action. In-depth study and understanding of immunotropic mechanisms (e.g., regulation of phagocytosis) is a prerequisite for adequate pharmacotherapy in infectious conditions, nonspecific inflammatory diseases, autoimmune and oncological disorders. The aim of our work is to study the effect of flavonoids upon phagocytic activity of professional phagocytes (neutrophils) using an in vitro test system. The biological material (venous blood) from 30 practically healthy people (adults n = 15, children n = 15) was used in the present work. The study was carried out in accordance with established international regulations. 0.5 mg/L of Luteolin (basic substance content ≥ 98%) and Quercetin (basic substance content ≥ 95%) were added to experimental samples and incubated for 20 min at 37 °С. The percentage of phagocytosis, phagocytic number (mean number of formalin-treated sheep erythrocytes engulfed per one neutrophil) was determined in control and experimental samples using light microscopy. The unidirectional nature of phagocytosis inhibition by quercetin and luteolin was noted in the test experiments. A statistically significant (p < 0.05) decrease in the phagocytosis intensity by 10% was shown in experimental blood samples obtained from adult patients compared with control values. When quercetin and luteolin were added to blood samples obtained from children, a statistically significant (p < 0.05) decrease in phagocytosis by 30% was noted against control values. At the same time, the mean percentage of phagocytosis and phagocytic number in blood samples after the addition of flavonoids were found to be in the range of reference values, thus suggesting adequacy and physiological suppression of excessive activities of innate immunity compartments by quercetin and luteolin. At this concentration, the flavonoids were found to exert a more pronounced suppressive effect on phagocytic activity in children. Modeling of immune response using the phagocytosis indices assayed in experimental in vitro models with neutrophils from practically healthy adults and children enables us to expand the knowledge of mechanisms underlying the immunotropic effects of flavonoids (quercetin and luteolin), in order to correct immunopathological conditions.

The E antigen of Ambrosia artemisiifolia (Amb a1) is the most potent ragweed allergen. In 97% of patients with ragweed pollen allergy, IgE antibodies to the Amb a1 are detected in blood serum, being associated with a positive skin prick test for the Amb a 1 allergen. In humans, the tryptase alpha/beta 1 (TPSAB1) enzyme is simultaneously released from mast cells resulting from contact of sensitized person with this allergen. Absence of tryptase inhibitor in humans is the typical feature of this enzyme. We have attempted to determine the most significant points of TPSAB1effects after its splitting into peptide fragments. Peptidase cleavage was carried out using the Bioscan 9.14 computer program ODO NICP Resan (Belarus), and the US National Center for Biotechnology Information (NCBI) database. The international one-letter amino acid sequence code was used for calculations and presentation of results. The data on peptide interactions with human proteins were obtained using the SwissTargetPrediction program. The test specimen was as follows: Ambrosia artemisiifolia antigen E (Amb a1) GenBank: AAA32665.1. Cleavage of the sample was carried out from position 1 to the last amino acid (No. 396). The length of split fragments is not specified. The studied enzyme was Homo sapiens tryptase alpha/beta 1 (TPSAB1) Gene ID: 7177, updated: 13-May-2022; enzyme type: endopeptidase. Split positions: 1 r|x and 2 k|x. The following amino acid sequence was analyzed: Ambrosia artemisiifolia antigen E (Amb a1) GenBank: AAA32665.1. It has been found that the E antigen from Ambrosia artemisiifolia (Amb a1) contains 396 amino acid residues. The 40 peptide fragments of the split sample were obtained. The ligand-receptor interaction was analyzed for peptides with a length of 2 to 4 amino acid residues, which had the strongest regulatory potential (p1-4 mgik, p56-57 gk, p127-129 ldk, p143-145 gak, p274-276 mpr). It has been shown that each peptide from the Amb a1 sequence acts as a ligand for specific receptors mediating the effects upon certain mechanisms in the patient’s body. Further study of these interactions enables identification of the most significant proteins (enzymes), which, upon impact of E antigen from Ambrosia artemisiifolia (Amb a 1) may lead to changed functional activity of regulatory systems in humans suffering from allergies.

Markers for identification of ER+/PR breast cancer (BC) risks and conversion of ER+/ER+ to ER-/PR- tumors are necessary for effective prevention and therapy of BC by the selective ER – modificators. Purpose – To reveal the proposed associations of gene polymorphisms of   ESR1 and ESR2 and antiidiotypic antibodies to estradiol (IgG2-E2) with ER+/PR+ BC risk and conversion of ER+/PR+ to ER-/PR- tumors and to study the interrelations between gene variants of ESR and IgG2-E2 in healthy women and BC patients. Polymorphic loci of ESR1 (rs2234693) and ESR2 (rs4986938) were studied by the real time PCR in 370 healthy women and 1169 BC patients. Tumor tissues ER and PR were detected by the standard immunohistochemical methods. Serum IgG2-E2 were studied using non-competitive enzyme immuno-assay. TT, TC and CC genotypes of ESR1 were revealed with the equal frequency in healthy women and BC patients I stage. Homozygotes GG of ESR2 were detected rarely, but AA frequently in BC patients with ER+/PR+ tumors at the I stage, than in healthy women женщин (44.0% and 14.2% vs 52.7 and 8.4%, respectively; p=0.005). The low levels of IgG2-ES were revealed more rarely but high levels more frequently in BC patients with ER+/PR+ tumors at the I stage, that in healthy donors (39.8% and 60.2% vs 58.0% and 42.0%, respectively; p=0.0002). Decreasing of ER+/PR+ tumors proportion and corresponding increasing of ER-/PR- tumors from I stage to II–IV stages (71.7% to 58.9% and 13.9% to 23.1%; p=0.006) were revealed in TC heterozygotes of ESR1 only. The same conversion of ER+/PR+ tumors to ER-/PR- were detected in GG homozygotes of   ESR2 (p=0.004). There have been the similar ER/PR transformation in BC patients with high IgG2-E2 levels (74.7% to 57.6% and 11.3%, to 25.0%, respectively, p<0.0001). High and low IgG2-E2 levels were detected with the same proportions at the any genotypes of ESR1 and ESR2 in healthy women or in BC patients. ESR1-2 gene polymorphisms and serum IgG2-E2 levels may be used as independent markers for prediction of ER+/PR+ and for ER+/PR+ to ER-/PR- tumors conversion during BC progression.

Diabetic retinopathy is a common complication of diabetes mellitus, especially, in elderly persons, due to growth of this population in many countries. However, involvement of immune system in patients with diabetic retinopathy and into the aging process is not sufficiently covered in the research works. The aim of the present study was to evaluate the contents of systemic interleukins in the patients with diabetic retinopathy with accelerated versus physiological aging.

We observed 240 patients aged 60-74 years with diabetic retinopathy and 115 age-matched patients without diabetic retinopathy under clinical conditions. The diagnosis of diabetic retinopathy was assessed in accordance with Clinical Guidelines of the All-Russian Association of Ophthalmologists “Diagnostics and Treatment of Diabetic Retinopathy and Diabetic Macular Edema”. The biological age of the subjects was determined instrumentally, by means of VaSera VS-1500 sphygmomanometer. The contents of interleukins in blood plasma was determined by ELISA technique using the “Protein contour” kit.

It was established that the chronological (calendar) age of patients with diabetic retinopathy and without diabetic retinopathy was not significantly different (70.9±0.7 and 70.2±0.8 years old, respectively; p > 0.05). However, the biological age in these groups differed significantly (75.7±1.1 and 72.3±1.0 years old, respectively; p < 0.001), thus suggesting accelerated aging of patients suffering from diabetic retinopathy. Plasma concentrations of interleukins in patients with diabetic retinopathy with accelerated aging, when compared with physiologically aged patients with diabetic retinopathy revealed statistically significant differences for the most analyzed interleukins. A particularly pronounced increase of the blood plasma interleukins in patients with diabetic retinopathy and accelerated aging was revealed for IL-6 (25.7±1.8 pg/mL versus 4.2±0.5 pg/mL in physiologically aged patients with diabetic retinopathy (p < 0.001). A significant increase of interleukin levels among patients with accelerated aging and diabetic retinopathy was found for IL-13 and IL-17. IL-13 contents in the patients with diabetic retinopathy and accelerated aging reached 2.2±0.3 pg/mL versus 0.7±0.2 pg/mL in physiologically aging patients with diabetic retinopathy (p < 0.001). Respectively, the IL-17 levels were 19.8±0.6 pg/mL and 8.4±0.9 pg/mL. The mean concentration of IL-1b, IL-3 among patients with diabetic retinopathy and accelerated aging was also significantly increased. At the same time, in the blood plasma of the prematurely aged patients with diabetic retinopathy, we have revealed a statistically significant decrease of anti-inflammatory interleukins and, especially, IL-10 to 7.4±0.6 pg/mL versus 19.2±0.7 pg/mL (p < 0.001). Therefore, IL-6, IL-8, IL-13, IL-17, IL-4, and IL-10 levels may be used as immunological predictors of accelerated aging in the patients with diabetic retinopathy.

The similarity of clinical manifestations and some pathogenetic mechanisms in rheumatoid arthritis (RA) and osteoarthritis (OA) is of particular interest in studying the features of extracellular neutrophil traps (NETs) formation in these musculoskeletal diseases. Our objective was to evaluate circulating neutrophil extracellular trap formation related to autoimmune inflammation in RA and reactive inflammation in OA.
39 RA patients comprised the main group; 35 OA patients, control group; 33 healthy individuals were included into the reference group. Circulating neutrophils were isolated by the one-step centrifugation in double layer of iohexol density gradient. NETs were induced by phorbol-12-myristate-13–acetate (PMA) being visualized by fluorescence microscopy.
The RA disease activity at the inclusion period should not exceed 2.6 DAS28 points. ОA patients were in clinical remission at the inclusion timepoint. In 17 RA patients, an increase in DAS28 exceeded 3.2 points; in 25 OA patients, an exacerbation was diagnosed during the study. The mean percentage of spontaneous and induced NETs in reference group was 3.8% (2.6-5.0) and 12.2% (9.0-15.4), respectively. Appropriate values for the patients with inactive RA were 5.9% (5.6-6.2) and 26% (23.9-28.1); 16.6% (16.1-17.1) and 38.0% (36.6-39.4) in active RA cases, respectively; 5.4% (5.2-5.6) and 20.3% (18.3-22.3) in OA without synovitis; 13.1% (12.6-18.1) and 28.3% (6. 4-30.2) in OA with synovitis, respectively. Spontaneous increase of NETs during RA activation was 181.4%; induced increase in NETs was 46.2%; and in cases of OA exacerbation these values were 142.6% and 39.4%, respectively. In RA patients, the rates of spontaneous NETs formation were 3.9 times higher than for induced values. Appropriate index was 3.6 times higher among the OA patients. The increase in spontaneous -to-induced NETs formation was more pronounced in active RA than in OA with synovitis. The main difference in NETs composition for active RA patients (in 88% of cases) and OA synovitis (in 50% of cases) is related to the contents of citrulline epitopes (p = 0.03).
Transition of RA from the remission state to active inflammation, as well as OA exacerbation are accompanied by a significant increase in NETs, especially, spontaneous formation. The degree of increase in RA was higher than in OA, thus, probably, indicating higher involvement of neutrophils in generation of NETs during autoimmune inflammation rather than in reactive inflammation. A sufficient role of citrullinated epitopes in NETs demonstrates their influence upon induction and maintenance of autoimmune response to RA-specific autoantigens.

Glomerulonephritis (GN) is a group of immuno-inflammatory kidney diseases with predominant glomerular lesions that are difficult to treat. The greatest problems are caused by the treatment of GN with nephrotic syndrome, which often has a recurrent course. The aim of the research was to study the effectiveness of recombinant interleukin-2 (rIL-2) therapy in the GN patients with nephrotic syndrome. 62 patients with a nephrotic form of primary GN with frequent relapses admitted to the Nephrology Department have been recruited into the study. The age of patients was from 18 to 65 years. The patients underwent standard examinations, as well as immunological studies, before administration of the anti-relapse treatment, and 12 months after the treatment was started. Immunological testing included immunophenotyping of lymphocytes with counting of T and B lymphocytes, immunoregulatory and activated subpopulations of T lymphocytes, determination of urinary immunoglobulins (IgM, IgG, IgA) by immunoturbidimetric assays, proinflammatory cytokines – IL-1β, IL-8, IL-17A and anti-inflammatory cytokine IL-10 by ELISA tests. As a result of studies, the examined patients showed an increased contents of T helper cells, activated T lymphocytes (CD8⁺HLA-DR⁺CD45⁺, CD3⁺CD8^brightCD38⁺) along with decreased numbers of Treg cells and an increased contents of proinflammatory cytokines IL-1β, IL-8, IL-17A and immunoglobulins of all three classes in urinary samples.

The cohort of patients with GN selected for the study was divided in two groups (the main group and the comparison group). In addition to nephroprotective and steroid therapy, the treatment regimen of patients included rIL-2 in the main group, or cyclophosphamide in the comparison group. Regardless of the method used, the levels of protein, IgG and IL-17A in the urine proved to be decreased relative to the initial values; the contents of B cells and HLA-DR⁺ cytotoxic T lymphocytes in peripheral blood were found to be decreased. The revealed changes were more pronounced in the main group of patients. By 12 months after starting the treatment, the mentioned indexes began to differ significantly in the main group from those in the comparison group. Serum creatinine levels, numbers of T helper cells and Treg cells, IL-1β levels in urine did not undergo significant changes in the comparison group, whereas a decrease in serum creatinine and urinary IL-1β was registered in the main group of patients, along with decreased number of T helpers and increased numbers of Treg cells. In the main group of patients treated with rIL-2, the average number of relapses per year decreased by 4 times, showing only a 1.2-fold decrease in the comparison group. Hence, the low-dose therapy with rIL-2 may be considered an effective and safe alternative to conventional immunosuppressive therapy and a new option of the targeted treatment of glomerulonephritis with frequent recurrence of nephrotic syndrome.

The worldwide circulating influenza viruses annually lead to serious medical and socio-economic consequences. It is generally recognized that vaccination is the most effective and safe strategy for preventing influenza and its complications. In order to reduce side effects when using live viruses, split and subunit influenza vaccines are widely used. To date, the characteristics of B cell response after immunization with influenza vaccines remain insufficiently studied. The aim of our study was to compare the effects of immunization with different influenza vaccines, i.e., “Sovigripp”, “Grippol plus” and “Ultrix”, on the B cell response. The study was conducted on the base of Clinical Department at the A.Smorodintsev Influenza Research Institute during the epidemic flu season of 2018-2019. For clinical studies, venous blood samples were obtained from 39 volunteers before vaccination, on the 7th and 21st days after vaccination. The subpopulations of B cells were analyzed by flow cytometry using fluorescently labeled antibodies to CD3, CD19, CD20, CD27, CD38, IgD, IgA surface antigens (BioLegend, USA). Cryopreserved mononuclear cells (1 × 10⁶ cells/sample) were used for analysis. The processing of flow cytometry data was carried out with special software (H., Cytexpert, Beckman Coulter, Inc., USA) and Kaluza 2.0 (Beckman Coulter, Inc., USA). The differences with pre-vaccination data were evaluated using the Mann–Whitney U-test and being considered significant at p < 0.05. As a result of the studies, the following subpopulations of B lymphocytes (CD3^-CD19⁺) were specified: naive B cells (CD20⁺CD27^-IgD⁺), non-switched memory B cells (CD20⁺CD27⁺IgD⁺), switched memory B cells (CD20⁺CD27⁺IgD^-), effector memory B cells (CD20⁺CD27^-IgD^-), plasmablasts (CD20^-CD38^hiCD27^hi). Activation of the B cell immune response was assessed by measuring the relative content of CD38⁺B cells belonging to subpopulations of naive, effector B lymphocytes, switched and non-switched memory B cells. The analysis of B cell response showed an increase in both the total number of B lymphocytes and their subpopulations including plasmablasts and activated switched memory B cells after immunization. With adjuvant vaccines “Grippol plus” and “Sovigripp”, as compared with the split “Ultrix” vaccine, an early increase in relative counts of plasmablasts was shown on the 7th day of the study. At the same time, all three vaccines equally contributed to an increase in the number of activated memory B cells with a switched antibody isotype. Thus, the assessment of B cell response revealed significant changes in contents of peripheral blood B cell subpopulations in response to vaccination with “Grippol plus”, “Sovigripp”, or “Ultrix”.

The main cause of edema in hereditary angioedema (HAE) is due to elevated bradykinin levels, caused either by C1-INH deficiency/change in functional activity and caused by mutations in the SERPING1 gene or by mutations in the F12, PLG, ANGPT1, KNG1, MYOF and HS3ST6 genes with a normal level and functionality of the C1-esterase inhibitor. The aim of the work was in silico prognostic analysis of the rare synonymous variant NC_000003.12:g.186725098T>C in the KNG1 gene and its impact on the development of HAE symptoms. The material was a whole blood sample obtained from a woman with clinical manifestations of hereditary angioedema without a decrease in the levels and function of the C1 inhibitor. The research methods included whole exome sequencing, bioinformatic analysis of the KNG1 gene mutation using a number of databases and web resources. Results. When processing full-exome sequencing data, we detected a synonymous variant in the KNG1 gene (exon 4, isoform 1): NC_000003.12:g.186725098T>C. The patient is a heterozygous carrier of the variant, with a frequency of 0.000004 (1:264690). Presumably, the identified variant can lead to the development of sporadic edema through several pathways that are associated with the formation of bradykinin or its analogues. Therefore, (1) the mutant high-molecular-weight kininogen is more easily activated by kallikrein and becomes a source of bradykinin formation through the kallikrein-kinin system; (2) the mechanism of bradykinin formation undergoes significant changes and results in the formation of functionally active but aberrant bradykinin, which alters its inactivation by enzymes with a consequent increase in its half-life, (3) the changes in positions 380-389 bring about modifications in Lys-bradykinin reproduction such that in subsequent steps it is “easily” cleaved to bradykinin by arginine aminopeptidase. The results of our study therefore indicate a possible role of the identified variant in the KNG1 gene in the development of HAE.

The results of study relationship between antigen reactivity of T-lymphocyte population under ex vivo conditions and the intensity of protective post-vaccination immunity to causative agent of brucellosis are presented. Тaking into account the peculiarities of immunopathogenesis brucellosis and prevailing role of adaptive T-cell immunity to protect against the causative agent of infection, possibility predictive evaluation of protective immunity against brucellosis using CAST-tests is considered as the most important aspect of brucellosis problems. There is an obvious need for an ex vivo correlation analysis of the activity of antigen stimulation of T cells and the intensity of protective immunity formed after vaccination. A close direct proportional relationship was established between the number of live microbial cells Brucella abortus 19BA vaccine strain administered and increase in ex vivo CD3-cell activation. A close correlation was revealed between ex vivo value of antigen-induced stimulation CD3-lymphocytes and level of post-vaccination immunological protection against brucellosis infection. It has been shown that in biomodels vaccinated against brucellosis with a T-lymphocyte stimulation coefficient of 50% or more (according to intensity of antigen-induced ex vivo expression CD25), 100% protection from the development of brucellosis infection after infection with Brucella melitensis at a dose of 1 × 10³ live microbial cells are provided. At the same time, there was a lack of a close correlation between an increase in the dose of brucella vaccine strain administered to biomodels and a change in geometric mean antibody titer, presence of a weakly pronounced relationship between level of agglutinins and immunological protection of biomodels from development brucellosis infection and indicators bacterial contamination body.Based on results of study, it was demonstrated that it is possible to quantify the formation and protective activity of T-cell immunity to causative agent of brucellosis based on analysis of level antigen reactivity of CD3-lymphocytes ex vivo. The data obtained and described methodological approach can be used as a predictive criterion in assessing protective level of cellular immunity to causative agent of brucellosis in vaccinated or recovering patients, as well as in order to analyze effectiveness of specific prophylaxis brucellosis and study immunogenicity and protective properties candidate for brucellosis vWe present the results of studies related to antigen reactivity of T lymphocyte population under ex vivo conditions and the intensity of protective post-vaccination immunity to causative agent of brucellosis. Due to peculiarities of immunopathogenesis in brucellosis infection and prevailing role of adaptive T cell immunity for protection against the causative agent of infection, a predictive evaluation of protective immunity against brucellosis using CAST-tests is considered the most important issue in the field. There is an obvious need for ex vivo analysis of correlations between the activity of antigen stimulation of T cells, and the intensity of protective immunity raised after vaccination. A close direct relationship was established between the number of live microbial cells of Brucella abortus 19BA vaccine strain administered, and increase in ex vivo CD3 cell activation. A close correlation (r = -0.841 ÷ -0.966, R2 = 0.708 ÷ 0.969) was revealed between ex vivo values of antigeninduced stimulation of CD3 lymphocytes, and the levels of post-vaccination immunological protection against brucellosis infection. We have shown that, in biomodels vaccinated against brucellosis with a T lymphocyte stimulation coefficient of 50% or more (according to intensity of antigen-induced ex vivo CD25 expression), 100% protection against brucellosis infection was achieved after contamination with Brucella melitensis at a dose of 1×10³ live microbial cells. At the same time, a lack of a close correlation was noted between an increased dose of Brucella vaccine strain administered to biomodels, and a change in geometric mean of antibody titer (R2 = 0.357÷0.404), along with a weak relationship between the levels of agglutinins and immunological protection of biomodels from developing brucellosis infection and indices of in vivo bacterial contamination.
These results suggest an opportunity to quantify development and protective activity of T cell immunity to the causal agent of brucellosis based ex vivo levels of antigen reactivity of CD3 lymphocytes. A correlation analysis between the state of T cell antigen reactivity and immunological resistance to brucellosis infection indicated a high degree of closeness between these indices. The key influence on activity of protective immunity is exerted by the levels of antigen reactivity of T lymphocytes, whereas the quotient of antigenic stimulation in CD3+CD25+ population may be considered the most informative index of immune protective activity. The data obtained and the described methodology may be used as a predictive criterion in assessing protective level of cellular immunity to causative agent of brucellosis in vaccinated or recovering patients, testing the efficiency of specific prophylaxis in brucellosis and studying immunogenicity and protective properties of candidate vaccines against brucellosis.