Vol 28, No 4 (2025)

Natural killer (NK) cells represent a critical component of antiviral immunity, demonstrating remarkable adaptability during infections such as human cytomegalovirus (hCMV) and SARS-CoV-2. Recent advances in single-cell transcriptomics have revealed the clonally expanding NK cell populations with distinct functional profiles, blurring the traditional boundaries between innate and adaptive immunity. However, the functional heterogeneity and immunological significance of these clones remain incompletely understood. Our aim was to dissect clonal NK cell heterogeneity using published scRNA-seq datasets from hCMV-seropositive, seronegative, and COVID-19 patients, focusing on cluster-specific gene expression patterns. Our computational pipeline employed Seurat-based integration and high-resolution clustering of datasets from hCMV-seropositive (n = 5) and seronegative (n = 2) donors, along with COVID-19 patients (n = 2). We analyzed datasets using Seurat 5 in R. Quality-controlled data were normalized (SCTransform), integrated (batch-corrected), and clustered (UMAP). Differential gene expression (Wilcoxon test, log2FC > 0.25, p-adj < 0.05) and annotation were performed. In hCMV-seropositive individuals, we identified 12 transcriptionally distinct NK cell clusters exhibiting KLRC2 (NKG2C)-dependent organization, with specific clones showing either enhanced cytotoxic potential (marked by GZMB/GZMA upregulation), or unique inhibitory receptor profiles (variable KIR expression patterns). The hCMV-seronegative cohort displayed a simpler clonal structure with 9 clusters showing reduced KIR diversity but maintained distinct effector gene signatures. Analysis of COVID-19 patients revealed divergent clonal patterns: one patient showed reduced KLRC2 variability with prominent KLRC1 (NKG2A) expression, while another exhibited KIR heterogeneity without KLRC2 variation. Our analysis reveals distinct clonal NK cell populations in hCMV and COVID-19 contexts, characterized by divergent expression of activating and inhibitory receptors. These findings demonstrate infection-specific dynamics of clonal NK cell populations, highlighting their adaptive potential through differential receptor expression in antiviral responses.

Targeting of adoptive immune cells is among modern approaches to therapy of solid tumors. Expression of an epidermal growth factor receptor HER2 is a common finding (20% of breast tumors) and is associated with a negative outcome. In this regard, obtaining HER2-specific effector cells carrying a chimeric CAR receptor is a relevant task. NK cells have a wide spectrum of activating receptors capable of recognizing tumor-associated markers and do not initiate a graft-versus-host reaction. The goal of this study was to obtain effector CAR-NK cells capable of eliminating the HER2-positive tumor targets. The NK cells were obtained by negative magnetic separation from peripheral mononuclear cells isolated from volunteers’ blood using density gradient centrifugation. Activated NK cells were modified by retroviral transduction. Preliminarily transfected Phoenix Ampho cells were used to accumulate retroviral particles carrying HER2- CAR construct. The DARPin 9-29-HER2 molecule with affinity for the HER2 distal domain I was used as the antigen-recognizing domain. The proportion of transduced NK cells was measured by means of GFP reporter protein expression. The surface emergence of HER2-CAR receptors was detected by expression of the c-Myc extracellular domain. All modified GFP⁺NK cells were capable of expressing HER2-CAR receptors on the cell membrane. Functional activity of HER2-CAR NK cells was measured using flow cytometry, by degranulation intensity and IFNγ production in the presence of HER2-positive target cells BT-474. To assess lytic activity of HER2-CAR NK cells, the cultures of HER2-CAR-expressing GFP⁺NK cells and unmodified GFP^-NK cells were obtained by cell sorting. Lysis of BT-474 targets was measured by calcein release upon incubation with HER2-CAR and GFP^- effectors. HER2-CAR NK cells were characterized by higher levels of degranulation and IFNγ production compared to GFP^-NK cells. Moreover, HER2-CAR-NK cells had higher lytic activity towards BT-474. Thus, by means of genetic modification based on primary NK cells, we obtained highly effective HER2-CAR-NK agents capable of HER2-positive target recognition and possessing cytotoxic and cytokine-producing potential in presence of tumor cells. Expanding the variety of cellular effectors aimed at treating HER2-positive breast tumors will increase the potential of personalized tumor therapy in the future.

Enriched habitat of laboratory animals causes stimulation of neuronal plasticity including increased level of hippocampal neurogenesis and more efficient spatial orientation. In addition, the enriched environment has an immunomodulatory effect on both peripheral immune cells and microglia, i.e., resident brain macrophages. Since immune cells and their secreted products modulate the processes of neuronal plasticity, studying the effects of enriched environment is relevant on the level of neurogenesis in rodents with different immune status. Compared with C57Bl/6, Balb/c mice are characterized by predominance of humoral (Th2) rather than cellular (Th1) types of immunity which is associated with differences in the H2 region of the major histocompatibility (MHCI) gene locus, as well as a higher level of anxiety revealed by behavioral phenotyping. The aim of our study was to evaluate the effect of enriched environment on spatial memory, levels of neurogenesis, and activation features of resident macrophages (microglia) in hippocampus of Balb/c mice. The study was performed on 4-month-old female Balb/c mice. To assess the spatial memory and learning ability, the Barnes labyrinth was used. Gene expression levels were assessed by means of gene-specific PCR using primers for BDNF, CD68, DCX, FGF2, IBA1, IL-1B, IL-10, SOX2, TMEM119, TNFα; GAPDH was used as a reference gene. The obtained data suggest that keeping Balb/c mice under the enriched conditions promotes increased levels of hippocampal neurogenesis, along with more efficient spatial orientation. Expression of genes encoding neurotrophic factors, as well as cytokines and markers of microglia activation proved to be increased for FGF2, TNFα, Iba1 and TMEM119, whereas expression of BDNF, IL-1β, IL-10 and CD68 genes did not change. Hence, expression of BDNF and IL-10 genes in Balb/c mice from enriched environment was not increased, being typical for C57BL/6 mice. However, it does not prevent the proliferation and differentiation of neurons in gyrus dentatus and successful spatial orientation in Barnes labyrinth. Microglial cells in Balb/c mice under enriched conditions are probably polarized according to the M0/M2 type.

Herpes simplex virus type 1 (HSV-1) is taxonomically classified as α-herpesvirus and is a complex double-stranded DNA virus. HSV-1 infections are extremely common among human population, with a global prevalence of 65% in persons under 50. In addition to labial and genital lesions, HSV-1-associated diseases include herpetic stromal keratitis, HSV encephalitis, and Alzheimer’s disease. Therefore, adaptive immune response is essential for controlling HSV infection, its reactivation, and complications. Key components of the adaptive immunity include IFNα and IFNβ, which play an important role at the early stages of infection caused by herpesviruses. Increased IFNα expression induces a systemic immune response, by activating NK cells, T lymphocytes, and increasing their migration to the inflammation site, as well as suppression of the viral life cycle by stimulating NK cell cytotoxicity and Th1 cell differentiation. Moreover, the main functions of IFNβ are to induce expression of interferon-stimulating genes (ISGs), whose expression products are able to inhibit viral reproduction cycle at various stages. A wide range of drug therapies and other approaches are currently used to treat herpes infections. Over time, however, an increased incidence of HSV-1 infection is observed in general population. In view of the current trends, it is necessary to search for new approaches aimed at reducing the incidence of HSV-1 infection and its complications. A promising approach may include usage of RNA interference effect, which underlies the action of potential new-generation antiherpetic drugs. RNA interference is a targeted inhibition of mRNA translation which entails disruption of subsequent protein biosynthesis. During its reproduction, HSV-1 imports viral DNA through the nuclear pore complex (NPC), located at the nuclear membrane. The NPC consists of nucleoporin proteins (Nup98, Nup205, NXF1 and others). Therefore, disruption of NPC structure resulting from miRNA-mediated inhibition of nucleoporin protein formation may hypothetically lead to a decreased HSV-1 reproduction.

Multiple myeloma (MM) is an incurable disease with common recurrence after treatment, including high-dose chemotherapy (HDC) followed by autologous hematopoietic stem cell transplantation (auto-HSCT). Tumor microenvironment is a factor of achieving long-term remission. Its immunosuppressive effects are mediated due to non-classical HLA class Ib molecules, i.e., HLA-E and HLA-G. These molecules interact with inhibitory receptors (NKG2A/CD94 for HLA-E; ILT2, ILT4, KIR2DL4 for HLA-G) on the surface of NK and T lymphocytes, blocking their cytotoxic activity and facilitating tumor evasion from immune surveillance. In contrast, the classical HLA-DR molecule plays an opposite role, enhancing antitumor immunity through the activation of CD4⁺T helpers. In this study, a comparative analysis of HLA-G/E/DR expression and counts of main immunocompetent cell populations (CD4⁺, CD8⁺, NK, NKT cells, B lymphocytes, monocytes) were performed by flow cytometry method in 10 patients with MM before and after HDC with auto-HSCT, and in 8 healthy volunteers. The results showed that, after therapy, the patients had a significant decrease of total CD3⁺T lymphocyte counts, including CD8⁺ and CD4⁺HLA-G⁺ subpopulations (p < 0.05), thus reflecting a cumulative immunosuppressive effect of melphalan-based conditioning. A decreased number of CD4⁺HLA-G⁺ cells, which have regulatory properties and are capable of suppressing the immune response, may indicate partial destruction of the tumor niche. At the same time, a paradoxically increased proportion of CD8⁺T lymphocytes, along with general lymphopenia, may be associated with clonal expansion of antigen-specific populations with “exhausted” T cell phenotype which requires further study. The results obtained suggest a dualistic role of HLA molecules in pathogenesis of MM: HLA-E/G act as key mediators of immunosuppression, while HLA-DR potentiates antitumor response. The discovered imbalance opens up new prospects for further studies of a role of HLA molecules in the tumor process, which may be applied to personalized therapy, including targeted blockade of NKG2A/LILRB1 receptors or HLA Ib molecules, and development of prognostic models based on a comprehensive analysis of HLA-G/E/DR expression.

Severe trauma remains a leading cause of disability and mortality among children over 1 year. This study aimed to assess the dynamics of peripheral blood lymphocyte subpopulations in pediatric trauma, depending on severity, complication risks, and general prognosis in pediatric trauma. A total of 118 children with severe mechanical trauma (ISS ≥ 16) were included. According to the Severe Injury Outcomes Scale (OISS), 60 patients had a favorable outcome; 51, with unfavorable outcome, and fatal outcome was documented in 7 cases. On the basis of clinical course, the subgroups with septic complications (PSC, n = 23) and multiple organ dysfunction syndrome (MODS, n = 19) were discerned. Flow cytometric analysis of lymphocyte subpopulations (CD3⁺, CD19⁺, NK cells) was conducted on days 1, 3, 5, 7, 10, and 14 post-trauma, with return to age-specific reference values. A comparison group included 34 children with mild or moderate trauma (ISS < 16), and a control group consisted of 60 age- and sex-matched healthy children. The laboratory findings revealed a significant and sustained reduction in CD3⁺, CD19⁺, and NK lymphocyte counts in children with severe trauma as early as day 1, with partial recovery by days 5-7. In children with mild trauma, these changes were less pronounced. Persistent lymphopenia, especially, in CD3⁺ and NK cells was significantly associated with MODS, PSC, and poor outcomes. These results confirm the prognostic relevance of dynamic monitoring of lymphocyte subpopulations as sensitive markers of immune dysfunction in the context of severe trauma and its complications. Inclusion of cellular immune parameters into risk stratification protocols may enhance the early identification of children at high risk for adverse outcomes and suggest targeted therapeutic interventions.

Specification of laboratory criteria for improving differential diagnostics of children with autoimmune hepatitis (AIH) and Wilson’s disease (WD) is a relevant task in pediatrics. Increased titers of antinuclear factor (ANF) in blood serum is among laboratory criteria for diagnostics of AIH. Some WD cases have been described which can manifest as autoimmune hepatitis (AIH) with detection of non-specific autoantibodies, thus complicating the differential diagnosis. The aim of our study was to identify the specific features for determination of antinuclear factor on the Hep-2 cell line and specific autoantibodies using a triple cell substrate in children with autoimmune hepatitis and WD. We have examined 62 pediatric patients with WD and 28 children with AIH. ANF was determined on the HEp-2 cell line (ANA-HEp-2, AESKUSLIDES^®, Germany). Specific auto-antibodies were detected by means of a triple substrate obtained from rodents, which included samples of kidney, liver and stomach tissues (LKS Mouse, Germany) using indirect fluorescence reactions applying an automated analyzer HELIOS^® (Germany). The analysis revealed ANF positivity in 15 out of 62 (24%) children with WD. Positive ANF results were obtained in 26 out of 28 children AIH (93%), being significantly more common in AIH compared with WD. The study of specific autoantibodies using a triple substrate revealed positive results in 34 children with WD (55%), versus 26 children with AIH (93%), Hence, the specific auto-antibodies on the triple substrate are more commonly found in children with AIH (p < 0.001) than in children with WD. In children with suspected AIH, when a homogeneous type of luminescence is detected on the HEp-2 cell line, and with a combination of different luminescence types on a triple substrate, the presence of Wilson’s disease in patients cannot be excluded.

Dysfunction of neutrophilic granulocytes (NG) is a decisive factor in the development and progression of atypical purulent-inflammatory diseases, including acute destructive pneumonia (ADP) in children. A promising approach is to include immunocorrective therapy in complex treatment, thus promoting recovery of NG functions. The objective of this study was to assess the features of defective NG functioning associated with changes in relative contents and phenotype of CD16⁺CD62L⁺CD11b⁺CD63^- and CD16⁺CD62L⁺CD11b⁺CD63⁺ neutrophil subsets in children with ADP, to evaluate clinical and immunological effectiveness of immunomodulatory therapy in complex postoperative treatment using a pharmaceutical drug with a synthetic thymic hexapeptide (pdHP) being the active substance. The study included 21 children aged 2-4 years: with a diagnosis of ADP (study group, SG) before complex treatment, and a group after treatment with hexapeptide (SG1). The comparison group included 20 conditionally healthy children (CG), Twenty children treated for ADP comprised the archival comparison group (aCG). The functions of NG were assessed by the content of CD16⁺CD62L⁺CD11b⁺CD63^- and CD16⁺CD62L⁺CD11b⁺CD63⁺ subsets using FC500 (Beckman Coulter) device. We also determined indices characterizing phagocytosis (%PAN, PN, PI, %D, ID), NADPH oxidase activity (%PPC, MCI) in the NBT test. We have found a phenotypic transformation of physiological subset to CD16^dimCD62L^dimCD11b^brightCD63^- NG with defective immune properties, a 3-fold increase in the content of NG subset with hyperactivated phenotype CD16^brightCD62L^brightCD11b^brightCD63^bright (p < 0.05), a defect in phagocytic function, but increased oxidase activity (p_{1, 2} < 0.05) in immunocompromised children with ADP. After complex treatment with the inclusion of hexapeptide, we observed normalization of NG effector functions associated with phenotype remodeling and restoration of the subpopulation ratio. Positive clinical dynamics was revealed when compared with aCG children receiving conventional therapy, i.e., a significant reduction in fever duration (|δ| = 0.98), respiratory failure (|δ| = 0.99), more rapid regression of pleural effusion (|δ| = 0.96) and early removal of drainages (|δ| = 1.00). The obtained results indicate a pronounced positive clinical and immunological efficiency of pdHP usage in complex treatment of children with ADP.

The issues of widespread psoriasis are quite urgent, due to its high prevalence of this disorder, absence of a single etiopathogenetic concept, diversity of clinical forms and systemic nature of the disease manifestations. Metabolic disorders and biochemical defects play an important role in pathogenesis of psoriasis and may be associated with impaired lipid metabolism. High frequency of dyslipidemia and metabolic syndrome in psoriasis determine high risk of cardiovascular diseases. The study of changes in differential leukocyte counts upon general blood testing and biochemical parameters of peripheral blood in psoriasis is of great clinical and diagnostic importance. The purpose of the study was to evaluate basic laboratory parameters of peripheral blood in patients with psoriasis who underwent inpatient treatment at the all-day Department at Chelyabinsk Regional Clinical Dispensary for Dermato-Venereal Diseases. Laboratory parameters of general and biochemical blood tests were studied in 750 patients with psoriasis admitted to the Dispensary. The severity of psoriasis and clinical PASI index were assessed. The counts of red blood cells, leukocytes, platelets, and hemoglobin were determined in blood as well as RBC color index, erythrocyte sedimentation rate (ESR). We have also assessed the levels of glucose, direct bilirubin, total bilirubin, urea, creatinine, C-reactive protein, rheumatoid factor, cholesterol, and triglycerides in blood serum. The study revealed an increased ESR in elderly patients with severe psoriasis. Dyslipidemia with increased level of cholesterol and triglycerides in the peripheral blood was detected in middle-aged and elderly women and men with moderate-degree psoriasis. In patients with psoriasis over 45 years of age with moderate course of the disease, an excess of total bilirubin was more often observed. An increase in systemic inflammation markers, i.e., C-reactive protein, ESR, direct and total bilirubin is observed in psoriatic disease. Lipid metabolism disorders with increased cholesterol and triglyceride levels were more often observed in moderate course of psoriasis among middle-aged and elderly patients.

Hyperactivity of angiopoietin-like proteins (ANGPTL) involved in persistence of chronic inflammation may lead to progression of different arthropathies. The purpose of the study was to evaluate interrelations between serum angiopoietin-like proteins types 2, 3 and 4 with clinical and laboratory characteristics of osteoarthritis (ОА) and comorbid conditions. We have examined 42 OA patients (females, 78.6%; aged 59.2±8.7 years; mean disease duration of 8.9±5.5 years) More severe joint damage (assessed as radiological stage) was accompanied by an increased levels of ANGPTL type 4 (p = 0.014). The group of OA patients with synovitis had significantly higher rates of type 2 and type 4 ANGPTL than patients without synovitis (p = 0.039 and p = 0.021, respectively). The content of type 4 ANGPTL was also significantly higher in patients with increased values of ESR and C-reactive protein (CRP) (p = 0.018). The associated co-morbidities in OA patients were as follows: obesity (40.5%), arterial hypertension (AH) (38%) and features of metabolic syndrome (MetS) (31%). The levels of type 3 ANGPTL in patients with MetS was higher than in OA patients without MetS (p = 0.024). The levels of type 4 ANGPTL in obese patients was higher than in OA patients with normal body weight (p = 0.027), and the contents of type 3 ANGPTL in this group did not reach the level of statistical significance (p = 0.052). The dispersion analysis also showed a significant effect of obesity degree on the level of type 4 ANGPTL (p = 0.034). The levels of ANGPTL types 2, 3 and 4 did not differ between the groups of OA patients with/without AH (p = 0.11, p = 0.055 and p = 0.09, respectively). However, when comparing patients with different degrees of AH, the difference in ANGPTL type 3 levels reached the level of statistical significance (p = 0.049). In summary, ANGPTLs showed different dependence on the comorbid background in patients with OA: type 2 ANGPTL is associated with inflammatory processes in joints; type 3 ANGPTL correlates with presence of MetS and severity of AH. Type 4 ANGPTL is associated with obesity, inflammation and severity of joint damage.

Traumatic brain injury (TBI) is among the most common forms of neurological pathology. In traumatic brain injury, axonal damage occurs not only as a result of direct cytotoxic interactions, but also due to activation of pro-inflammatory cytokines and chemokines, which play a crucial role in development and maintenance of neuroinflammation. At the early terms after brain trauma, there occurs both activation of microglia, and formation of a pool of T cell subsets able to produce various chemokines. The aim of this study was to assess the contents of chemokines (CXCL8/MIG, CXCL9/IP-10, CXCL10/I-TAC) in cerebrospinal fluid (CSF) of patients with brain contusion of varying severity within the first 24 hours after hospital admission. The study included 86 patients diagnosed with “TBI: brain contusion,” who were divided into three groups based on the injury severity. A comparison group consisted of patients with brain concussion (n = 24). Obtaining CSF from healthy individuals is challenging due to absence of clinical indications for lumbar puncture. Chemokine concentrations (pg/mL) were measured by means of multiplex analysis based on xMAP technology (Luminex, USA), using Milliplex MAP test systems (Millipore, USA). The results showed a trend toward increased chemokine levels correlating with more pronounces severity of brain injury. Notably, CXCL9/MIG levels were elevated in all patients with brain injury (p < 0.05), unlike CXCL8/IL-8, which showed significant increases only in the 4^th group (severe TBI), and CXCL10/IP-10, which was significantly elevated in both the 3^rd (moderate TBI) and 4^th groups. Specifically, in the 4^th group (severe TBI), levels of CXCL8/IL-8 were increased 3.5 times, CXCL9/MIG showed a 9-fold increase, and CXCL10/IP-10 was 3 times higher than in control group (1^st group, mild TBI). To evaluate the diagnostic value of these changes, ROC analysis was performed, establishing a clinically significant chemokine thresholds that serve as highly informative markers of neural tissue damage. In conclusion, assessment of CXCL8, CXCL9, and CXCL10 levels in CSF following brain contusion provides valuable insights into their role in disease progression, i.e., recruiting T helper cells and neutrophils from peripheral blood into neural tissue and sustaining neuroinflammatory processes.

The COVID-19 pandemic caused by the SARS-CoV-2 virus has resulted in global morbidity and high mortality rates worldwide. According to case histories, for a long time after acute COVID-19 infection (six months to 2-3 years), patients may experience a pronounced fatigue, increased tiredness, higher incidence of acute respiratory viral infections per year, increased recurrence of skin diseases, allergies, exacerbation of pulmonary disorders, urinary tract diseases, increased recurrence of chronic viral diseases, e.g., herpesvirus and papillomavirus infections, aggravation of chronic cardiovascular and other somatic diseases of various organs and systems. Patients were examined at early as six months after recovery from acute COVID-19. Such persistent post-infectious consequences are referred to as post-COVID syndrome. When assessing post-COVID syndrome, it is necessary to identify the main clinical syndromes of multiorgan pathology characteristic of post-COVID patients. Endocrine and cardiac manifestations of post-COVID syndrome may be a consequence of direct damage from viral infection, immunological and inflammatory damage, as well as iatrogenic complications. Objective of our study was to assess the impact of acute COVID-19 severity on the course of post-COVID syndrome. The research objectives were as follows: 1. To analyze the severity of clinical manifestations of cardiovascular disorders in post-COVID patients depending on the degree of lung damage in the acute period of COVID-19 (CT0 to CT4). 2. To analyze the severity of clinical manifestations of endocrine disorders, including newly diagnosed pathology, in post-COVID patients depending on the degree of lung damage in acute period of COVID-19. Since we did not found any significant gender-and age-dependent differences, all patients were divided into groups by the degree of lung damage during acute period of COVID-19, according to clinical guidelines for the diagnosis and treatment of a new coronavirus infection. This study showed that the clinical picture of post-COVID syndrome is characterized by a pronounced diversity in development of multiple organ pathology, both newly diagnosed, or manifesting by increased frequency of exacerbations of chronic diseases. Conclusions: 1. According to the data obtained, reliable differences were obtained between groups CT0 and CT1-2, as well as CT0 and CT3-4: the frequency of exacerbating cardiovascular disorders in the post-COVID period is significantly higher in groups with lung damage over acute period of COVID-19 compared to the group of patients without lung damage. These data suggest that cardiovascular disorders are directly related to the severity of COVID-19, viral load, and were detected most frequently (68%) in post-COVID patients who initially had a severe coronavirus infection. 2. According to our results, the frequency of glucose metabolism disorders, including those detected for the first time, proved to be significantly increased in the post-COVID patients with lung damage during acute infection. Meanwhile, no significant differences were found for thyroid disorders, except of autoimmune thyroiditis (AIT). These disorders may be associated with usage of corticosteroid therapy in acute period of coronavirus infection, or with impairment of regulatory mechanisms of the endocrine and immune systems induced by SARS-CoV-2 virus, thus again confirming our assumptions concerning development of multiple organ pathology in post-COVID patients.

Polycystic ovary syndrome (PCOS) is among the most common endocrine disorders observed in women of reproductive age and is characterized by a wide range of clinical manifestations, including ovulatory dysfunction, hyperandrogenism, and metabolic alterations, such as insulin resistance and obesity. In recent years, increasing attention has been given to immune factors and systemic inflammation in pathogenesis of this syndrome. Our objective was to determine the levels of anti-Müllerian hormone, gonadotropins, proinflammatory cytokines, carbohydrate metabolism parameters, and insulin resistance index in women with different clinical phenotypes of PCOS depending on the type of menstrual cycle. The study included 86 women at their reproductive age, being divided into four groups: a control group (women without menstrual cycle disturbances) and three groups of patients with PCOS, stratified by the type of menstrual irregularity: regular cycle, oligomenorrhea, and amenorrhea. All participants underwent evaluation of serum levels of anti-Müllerian hormone, follicle-stimulating hormone, luteinizing hormone, glucose, insulin, and proinflammatory cytokines including IL-6, TNFα, and IL-17A. Insulin resistance index was also calculated in accordance with a homeostatic model. The findings demonstrated significantly increased levels of all evaluated hormonal and immune parameters in women with PCOS compared to the control group, with the most pronounced changes observed in amenorrheic subgroup. The results of the study confirm a systemic interrelationship between inflammatory, hormonal, and metabolic disturbances in women with PCOS. Proinflammatory cytokines may serve as potential biomarkers of disease severity, and their comprehensive assessment, along with hormonal and metabolic markers may enhance the accuracy of diagnosis and support a more individualized approach to therapy in this patient population.

Globally, the prevalence of bronchial asthma (BA) alone reaches 19%, with the number of affected individuals showing an upward trend. Atopic dermatitis (AD) affects 25-30% of children and 7-10% of adults. Remission remains elusive even with advanced genetically engineered biological therapies. Consequently, research into immunopathogenic pathways and the development of novel therapeutic strategies remain critical. A typical representative of “non-classical” HLAs, the HLA-E molecule, is a perspective for study. These molecules are also expressed across various tissues, notably in the respiratory system (bronchial epithelium) and skin keratinocytes. As a cell surface protein, HLA-E participates in diverse immune response pathways. VL9 peptides stabilize HLA-E expression, enabling presentation to its primary receptors, NKG2, on natural killer (NK) cells. In adaptive immunity, HLA-E serves as a ligand for CD8⁺ cytotoxic T cell receptors (TCRs). This study aimed to evaluate HLA-E-bearing cells (CD4⁺, CD8⁺, CD14⁺) in patients with BA and AD, both before and 12 months after initiating genetically engineered biological therapy (GEBT). Peripheral blood mononuclear cells (PBMCs) were analyzed from healthy donors (n = 16), BA patients (n = 4), and AD patients (n = 5). AD patients received dupilumab (300 mg loading dose), while BA patients were treated with benralizumab (30 mg). Prior to therapy, both AD and BA patients exhibited lower proportions of HLA-E-positive CD4⁺ and CD8⁺T cells compared to healthy donors. No significant differences were observed in HLA-E expression on CD14⁺ monocytes. During treatment, HLA-E levels across all cell types in patients reached levels comparable to those in healthy donors. These findings suggest HLA-E’s involvement in disease pathogenesis and the modulating effects of GEBT on HLA-E-positive cell populations.

Fever is an extremely common symptom in children and the most common reason for visiting a pediatrician. There are many studies worldwide indicating that physicians and parents have a lot of misconcepts and conflicting results regarding fever. This is due to the methodological limitations of available studies in the field of fever, some disagreements in terminology, orientation towards patient comfort when making treatment decisions and lack of a holistic approach to the body reactivity. In this study, we sought to identify the knowledge, attitudes and misconcepts of primary care physicians regarding fever in children. This cross-sectional study was conducted in June-July 2024 with the participation of physicians and nurses at pediatric clinics (n = 209). Physicians, with verbal consent, independently completed the proposed questionnaire. In the study, a third of respondents had conflicting answers, whether a fever state should be considered a normal or pathological response in health disorder. All pediatricians recommend to bring the temperature down, and only 13.4% preferred to do it, if the child feels unwell. The overwhelming majority believes that fighting fever is not harmful to the body, although 72.2% note that administration of antipyretics may mask more serious problems. It is positive that 58.8% of doctors suggest bringing down the fever at > 39-39.5 °C, as currently recommended. A fewer number of respondents (13.9%) associate fever with a bacterial infection. There are misconcepts among health workers regarding fever after vaccination, during teething, the needs for physical cooling measures, mandatory watering, and indications for seeking medical care. All health workers have fears regarding the consequences of pyrexia, especially regarding convulsions, loss of consciousness, protein denaturation, and dehydration. On the one hand, this attitude demonstrates alertness and encourages a diagnostic search. On the other hand, it increases fear of fever. As a result, the instructions and definitions given to parents regarding fever treatment are often contradictory and incomplete. Most health care providers recognize the lack of knowledge in this area. By focusing on fever treatment, we suggest an impression for parents that fever is harmful and antipyretics are beneficial. Are there long-term consequences of such a daily struggle when the primary goal is to maintain health?