Introduction. At the beginning of the SARS-CoV-2 pandemic, specialists in biomedicine were predominantly focused on development of preventive vaccines. However, there is a huge unmet need for specific ways of treatment. In this study we describe the development and testing new antiviral drug which acts through RNA interference mechanism.

The aim of the study was to evaluate the antiviral effect of siRNA against SARS-CoV-2 both in vitro and in vivo, and development of specific siNRA based therapeutic drug.

Material and methods. Bioinformatics methods and specialized software were used for the design of siRNA molecules. More than 15,000 siRNAs variants were designed, 13 variants of siRNAs were selected for synthesis. A system based on vectors expressing luciferase reporter genes fused with SARS-CoV-2 genes was used for screening their activity in vitro. In addition, an in vitro SARS-CoV-2 infection model was used. To improve stability, the most active siRNA variant, siR-7, was modified using locked nucleic acids (LNA), and the modified variant was designated as siR-7-EM. Dendrimeric peptide KK-46 was used for intracellular delivery of siRNA. The siRNA molecules were mixed with this peptide at a ratio of 1/20 by mass, and the resulting complex (siR-7-EM/KK-46) was added to the culture of infected cells or used for inhalation in a Syrian hamster infection model.

Results. As a result of screening a library of designed siRNA molecules, the most active variant – siR-7 – was identified, which targets a region of the virus genome encoding the essential RNA-dependent RNA polymerase (RdRp) enzyme. We demonstrated that LNA modification of this siRNA significantly improved its stability to degradation and enhanced its antiviral activity in vitro. In vivo experiments showed that the inhalations with siR-7-EM/KK-46 complex of infected Syrian hamsters resulted in a reduction in viral load in the lungs and a decrease in virus-induced inflammation.

Conclusion. A therapeutic strategy for the treatment of COVID-19 based on the inhalations with a complex of siRNA molecules suppressing the virus replication enzyme and a carrier peptide is proposed. The antiviral activity of the complex has been demonstrated on experimental models in vitro and in vivo. MIR 19, the world’s first siRNA-based drug specifically suppressing SARS-CoV-2 replication, was developed on the basis of the siR–7-EM/KK-46 complex.

Introduction. In connection with the threat of coronavirus infection, increased the need for the development of fundamentally new antiviral drugs with increased efficiency and availability. One of the promising directions in the development of such drugs is the use of inhibitors of viral reproduction based on small interfering RNA (siRNA).

Aim – to study the safety of MIR 19^® (siCoV/KK46) in single and multiple doses, as well as to study the efficacy and choice of dosage of MIR 19^® in the treatment of hospitalized patients with infection caused by SARS-CoV-2 (COVID-19) who do not require treatment in the intensive care unit.

Material and methods. Phase I clinical trial, the safety of MIR 19^® was studied for single and multiple administration. As safety criteria, the incidence of any adverse events, the incidence of serious adverse events according to complaints, the results of a physical examination, the results of assessing heart rate and blood pressure, respiratory rate and body temperature, as well as laboratory monitoring data – indicators of clinical and biochemical blood tests, general urine analysis. Also, for the registration and monitoring of AEs, the instrumental research methods – ECG and spirography were used. An open-label, randomized, controlled, multicentre, phase II study (NCT05184127) was then conducted to evaluate the safety and efficacy of inhaled MIR 19^® (3.7 and 11.1 mg/day: low and high dose, respectively) compared with standard treatment (group of comparison) in patients with coronavirus infection / COVID-19 who did not require mechanical ventilation at the time of inclusion in the study (n = 52 for each group).

Results. Based on the results of the phase I clinical trial, it can be concluded that the single and multiple administration of the MIR 19^® preparation is safe in healthy volunteers. Phase II clinical trials showed that patients in the low-dose group achieved the primary endpoint of simultaneously achieving fever reduction, respiratory rate normalization, cough reduction, and blood oxygen saturation > 95 % within 48 h, significantly earlier (median 6 days) than patients in the group of comparison (median 8 days). In the high dose group, no greater clinical efficacy was observed in relation to the group of comparison. None of adverse events were associated with MIR 19^®.

Conclusion. MIR 19^®, an etiotropic drug specifically suppressing SARS-CoV-2 replication, is safe with single and multiple use, is well tolerated and significantly reduces the time to clinical improvement in patients hospitalized with moderate COVID-19, compared with standard therapy in a randomized controlled trial. MIR 19^® is registered (registration certificate LP-007720), it is included in the Temporary Methodological Recommendations of the Ministry of Health of the Russian Federation «Prevention, diagnosis and treatment of new coronavirus infection (COVID-19)» and is actively used in clinical practice.

Introduction. The consequences of COVID-19 infection in TB patients remain unclear. Coronavirus infection caused by primary variants of SARS-CoV-2 burdened the course of TB in elderly patients but did not exacerbate TB in children. The review of scientific literature did not reveal any studies describing T- and B-cell specific immune responses to SARS-CoV-2 in children and adolescents with TB.

Aim – to study expressiveness and duration of cellular and humoral immune responses to SARS-CoV-2 in children and adolescents with pulmonary TB after coronavirus disease or exposure to coronavirus infection caused by omicron strain.

Material and methods. We carried out a cohort prospective study including 34 patients aged 2–17 years with different forms of pulmonary TB, received treatment in the Children and Adolescents’ Department of the Central TB Research Institute. The children had a history of either coronavirus disease or exposure to coronavirus infection (omicron strain). Laboratory diagnosis of coronavirus infection in patients with clinical signs of acute respiratory viral infection (ARVI) was based on detection of SARS-CoV-2 RNA in nasopharyngeal swabs (omicron strain). Cellular immunity was assessed using T-SPOT.COVID (Oxford Immunotec, Great Britain) on panel A (COV-A) (Spike antigens) and panel B (COV-B) (Nucleocapsid antigens). IgM and IgG antibodies against nucleocapsid protein were determined (ARCHITECT, Abbott, USA) with assessment of positivity ratio (PR). The results of qualitative signs were expressed in absolute numbers with indication of proportions (%), the calculation of statistical significance used Pearson’s χ² criterion. The comparison of T cell and humoral immunity parameters after one and three months used Wilcoxon T-criterion.

Results. In all cases coronavirus infection had a mild course, the clinical picture of ARVI was observed in 67.6 % (23/34) of cases, and SARS-CoV-2 RNA was only determined in 74 % of cases. IgM antibodies were absent in all cases one month after the disease initiation/exposure. IgG antibodies were detected in 76.5 % of cases one month after the disease/exposure and 53 % of cases 3 months after the disease/exposure (р < 0.01). Positive results of specific T-cell immunity were observed in 97.1 % of cases after one month and 79.4 % after 3 months. At the same time specific T-cell immunity (panel A and panel B) significantly decreased by the 3^rd month of our study (р < 0.01). Trends in changes in T-cell immunity parameters (panels A and panels B) in groups of patients with and without clinical manifestations of the disease were different. If in the group with clinical manifestations, the number of spots did not change significantly when comparing the results after 1 and 3 months observation, in the group without clinical signs, a statistically significant decrease was noted by the 3^rd month in panel B, in the absence of significant dynamics in panel A.

Conclusion. In our study, coronavirus infection in children and adolescents with tuberculosis in all cases proceeded in a mild form and was characterized by the presence of a pronounced specific T cell response in 97.1 % and the production of specific IgG-antibodies in 76.5 % of patients after 1 month of the study with subsequent statistically significant decrease in indicators by the 3^rd month of observation – 79.4 and 53 %, respectively.

Introduction. The participation of cytokines in reactions of adaptation and the formation of comorbid somatic at various stages of stress reaction is beyond doubt. The duration of preservation of cytokine imbalance can serve as a marker for assessing the state of reserve mechanisms of homeostasis in military personnel – participants in combat operations suffering from comorbid pathology.

Aim of the study – studying the ratio of pro- and anti-inflammatory cytokines in chronic stress-induced comorbid pathology in combatants in the long-term follow-up period.

Material and methods. 100 reserve officers were examined, divided into 2 groups: group I – military personnel with arterial hypertension, which first debuted while participating in hostilities on the territory of the Chechen Republic (1994–2001), and group II – military personnel with arterial hypertension, but not taking part in hostilities. The content of cytokines: interleukins (IL)-4, -6, tumor necrosis factor (TNF)α, interferon (IFN)-α и -γ – in blood serum was assessed in various variants of the course of comorbid stress-induced pathology in the long-term follow-up period.

Results. In the long-term follow-up period, reserve officers suffering from stress-induced comorbid pathology registered a continuing increase in the content of IL-6, as well as the preservation of increased the levels of TNFα and IFN-α.

Conclusion. In the long-term follow-up period, reserve officers suffering from stress-induced comorbid pathology retain cytokine imbalance, which is manifested by an increase in the content of IL-6 and an increase in the levels of TNFα and IFN-α.

Introduction. The development of autoimmune thyroiditis (AIT) is determined by defects in the immune system in individuals with a genetic predisposition.

The aim of our study was to assess the levels of organ-specific autoantibodies – thyroperoxidase antibodies (TPO-Ab), thyroglobulin antibodies (TG-Ab) and non-organ-specific autoantibodies – double-stranded DNA (Ab-dDNA) and single-stranded DNA (Ab-sDNA) antibodies in patients with various forms of autoimmune thyroiditis; analysis the correlation between the level of antibodies to DNA and the level of antibodies to thyroperoxidase, depending on the functional state of the thyroid gland.

Material and methods. Non-randomized study was performed, in which a comprehensive examination of blood serum samples from 58 patients (24 men and 34 women) with autoimmune thyroiditis, aged 18 to 64 years and 32 healthy persons (group of comparison, 14 men and 18 women) without thyroid pathologies and other autoimmune diseases aged 20 to 65 years, was conducted. Patients were equally divided into 2 groups: a group of patients with subclinical form of the disease (subclinical hypothyroidism) and a group of patients with a manifest form of the disease. In patients with a subclinical form of the disease, some mild manifestations of hypothyroidism were noted. All study participants were tested for the content of Ab-dDNA and Ab-sDNA in the blood serum by enzyme immunoassay using an automatic analyzer «EL 808 Bio-Tek Instruments, Inc.» (Diagnostic Products Corporation, USA).

Results. The levels of antibodies to DNA were higher in patients vs the group of comparison. The median concentration of Ab-dDNA in patients with AIT was 4.4 (2.3; 9.9) U/ml. It was statistically significantly higher compared to the group of comparison 2.6 (1.5; 3.6) U/ml (p < 0.001). There were no significant intergroup differences in the levels of Ab-sDNA in patients with various forms of hypothyroidism.

Conclusion. The study of the levels of IgG antibodies to DNA in the blood serum samples of patients with AIT depending on the functional state of the thyroid gland and the TPO-Ab level can be a prognostic marker and be used as an additional criterion for the development of an autoimmune process. According to the results of our study, the level of Ab-dDNA in the blood serum samples of patients with AIT positively correlates with the level of TPO-Ab.

Minireview addresses the question of whether passive immunization with palivizumab and routine vaccination can be combined in young children, including preterm infants. This question is increasingly coming from neonatologists and pediatricians and addressed to clinical immunologists and pharmacologists. Palivizumab is a humanized therapeutic monoclonal antibody used to prevent respiratory syncytial virus (RSV) infection. The drug belongs to the class of targeted monoclonal antibodies, which means its high selectivity for only RSV and only for one protein of this virus that provides virulence and is responsible for fusion with the infected host cell (F-protein). There are no direct indications of the possibility of combining anti-RSV prophylaxis and vaccination in the summary of product characteristics for palivizumab. However, based on the precisely described mechanism of action and according to all the canons of clinical immunology and pharmacology, palivizumab therapy cannot change the efficacy and safety of vaccination. For more than 25 years of clinical use of palivizumab, extensive experience has been accumulated, which is reflected in international and national clinical guidelines. The final conclusion of these recommendations to date: the combination of passive and active immunization is justified and recommended.

Introduction. A high degree of hypersensitivity to honey bee venom, high rates of anaphylaxis when stung by a bee (in the Russian Federation up to 6.4 %), the absence of therapeutic drugs in a wide allergological practice that allow the only effective and reliable method of treatment and prevention – allergen-specific immunotherapy – for patients with insect allergy to bee venom, which allows to reduce or completely eliminate the risk of anaphylactic type reactions, including anaphylactic shock with a fatal outcome, indicate the urgency of the problem of creating medicinal products.

The aims of the study – development of technology for obtaining and experimental evaluation of the bee venom allergoid.

Material and methods. The raw bee venom was purified, preparative and analytical chromatography and electrophoresis in polyacrylamide gel were performed. The gas chromatography technique was used for the analysis of impurities. A competitive enzyme immunoassay (EIA) was performed to determine specific circulating IgE antibodies in experimental animals and human.

Results. An allergoid was made from purified bee venom. Analysis of the binding of purified bee venom, allergen and allergoid in competition with biotinylated bee venom allergen in competitive reverse EIA showed that allergoid, as a result of formaldehyde treatment, lost majority binding sites for specific IgE antibodies to bee venom proteins. Evaluation of the immunogenicity of the allergoid using EIA, showed that in the sera of 104 immunized male mice 1, 2, 3 and 4 weeks after the start of immunization of animals, circulating specific IgG antibodies are determined and a gradual increase of its level is observed.

Conclusion. The developed allergoid has a high ability to induce the specific IgG antibodies, which indicates the possibility of its practical application for allergen-specific immunotherapy.

Introduction. The pharmacological effect of human immunoglobulin (HI) antirhesus Rh_o(D) drugs is characterized by their specific activity – the ability of anti-D antibodies (Ab) IgG to selectively block an undesirable immune response to the D antigen of Rh-positive fetal erythrocytes that have entered the mother’s bloodstream, as well as the dose of the administered. The dose of the drug is calculated based on the quantitative content of anti-D-IgG-Ab, expressed in International Units (IU) or in micrograms (mcg). Foreign manufacturers the content of anti-D-IgG-Ab in preparations determined quantitatively by hemagglutination, flow cytofluorimetry and enzyme immunoassay in IU or mcg (5 IU = 1 mcg) in relation to the International Standard (IS) of anti-D immunoglobulin or a standard sample (SS) calibrated relative to the IS. Domestic manufacturers determined the content of anti-D-IgG-Ab in the titers using an indirect Coombs method without using a SS. The absence of a national SS certified in IU (or mcg) did not allow domestic manufacturers of HI anti-rhesus Rh_o(D) drugs to establish the norm of anti-D-IgG-Ab content in the drug in IU (or mcg) to calculate the dose of administration in accordance with international requirements for its clinical use, which could cause the absence of the desired therapeutic effect, erythrocyte hemolysis or the development of other adverse reactions.

The aim – to develop a Pharmacopoeia Standard (PhS) for the quantitative determination of the activity of HI antirhesus Rh_o(D).

Material and methods. Samples of the candidate for PhS, IS anti-D immunoglobulin, SS of the European Pharmacopoeia with the content of anti-D-IgG-Ab determined in the agglutination reaction (1:8), human erythrocytes of the phenotypes R1R1 and R2R2, rr were used in study. To establish the certified characteristics of the PhS, the enzyme immunoassay and the hemagglutination in gel were used.

Results. The national PhS 3.1.000452 of the State Pharmacopoeia of the Russian Federation «Standard sample for determining the activity of IH antirhesus Rh_o(D) (content of anti-D-IgG-Ab)» has been developed with an activity value of 516 IU/ml (103 mcg/ml) and an extended uncertainty: ±100 IU/ml (±20 mcg/ml) (coverage ratio 2, confidence level 95 %). It has been established that the available validated enzyme immunoassay using erythrocytes of the R1R1 or R2R2 phenotypes for self-preparation of an immunosorbent provides an accurate quantitative determination of anti-D-IgG-Ab in the PhS. It is shown that the use of enzyme immunoassay and hemagglutination in gel to establish the certified characteristics of the PhS ensures the traceability and comparability of the results obtained.

Conclusion. The developed PhS is included in the register of PhS of biological origin, as well as implemented in the regulatory documentation for the drug produced in the Russian Federation (INN «Human Immunoglobulin anti-rhesus Rh_o(D)»). The use of PhS makes it possible to establish the values of the anti-D-IgG-Ab content in the drug in IU/ml (or mcg/ml), which contributes to an adequate calculation of the dose of administration during its clinical use for specific perinatal prevention of Rh-immunization of women with Rh-negative blood affiliation.

The prevalence of allergic diseases is getting higher. Patients of the Central District of the Russian Federation most often suffer from an allergy to a domestic cat: trigger proteins are found in dander and animal body fluids. 8 allergens of the domestic cat are known recently, however, in practice, native extract is used for the diagnosis of IgE-associated hypersensitivity, which contains almost all allergenic molecules. For a more personalized approach to treatment, an assessment of the individual molecular allergological profile of patients is necessary. We reviewed studies that examined the molecular characteristics of cat allergens and summarized the most valuable and clinically relevant data. Based on the review of the literature, we have concluded that it is necessary to continue the study of proteins, especially minor ones, since there is a shortage of information about their allergenic activity and clinical relevance. The most studied allergen is Fel d 1, as it is widely distributed among cat-sensitized patients. The specificity for determining the guilty cat allergen when using native allergens is low, which in turn reduces the effectiveness of the treatment. For a more accurate allergen-specific therapy, which is under active development, it is important to assess the molecular allergological profile of patients and standardize them for the feline allergen with which they associate the clinical picture of sensitization.

Chronic rhinosinusitis (CRS) is a group of diseases with different etiology and pathogenesis characterized by persistent inflammation of the mucous membrane of the nose and paranasal sinuses (PNS) and is subdivided into CRS without polyps, which is more prevalent (more than 2/3 of cases), and rhinosinusitis with nasal polyps (CRSwNP). Histologically, CRSwNP is characterized by mucosal infiltration of PNS with inflammatory cells (eosinophils, lymphocytes, plasmacytes, neutrophils, etc.), modifications in epithelial cell differentiation, tissue remodeling, including basement membrane thickening, epithelial degradation, oedema, and fibrosis of the submucosal layer with accumulation of extracellular matrix (ECM), basal cell dysplasia. Polarized epithelial cells transform into mesenchymal forms with loss of typical morphology, which are characterized by resistance to apoptosis, increased invasiveness and migratory ability, loss of polarity, adhesiveness, intercellular connections and hyperproduction of EMS proteins, the accumulation of which is critical for pathological reconstruction of the PNS mucosa. Metalloproteinases (MMPs), whose activation and biological activity is regulated by tissue inhibitors of metalloproteinases (TIMPs), are the main proteolytic enzymes responsible for extracellular matrix remodeling. The balance between MMPs and TIMPs determines the processes of production and degradation of individual components of the ECM, thereby influencing the pathogenesis of inflammation.

The current review summarizes the data of investigation of the role of ММPs in the upper respiratory tract remodeling during chronic rhinosinusitis. Experimental data on the molecular and cellular action mechanisms of ММPs were reviewed. The systematization of these data could help to reveal promising targets for the development of new approached for CRS treatment.