BACKGROUND: Asthma is considered one of the urgent medical and social problems of the XXI century. In recent years, not only an increase in the incidence, but also an increase in the prevalence of severe forms of asthma has been noted in all countries of the world. The effect of the sensitization spectrum on the severity and level of asthma control has not been studied enough. AIM: Evaluate the importance of sensitization spectrum to aeroallergens in severe asthma patients as indicators for disease exacerbation risk. MATERIALS AND METHODS: Single-center cross-sectional study at North-Western State Medical University n.a. I.I. Mechnikov. Examined 93 severe asthma patients. Analysis covered demographic traits, comorbidities, hereditary history, living conditions, exacerbations, past-year hospitalizations, medications, spirometry, and specific IgE presence for 10 aeroallergens. RESULTS: Aspergillus fumigatus specific IgE notably impacted asthma control. Disease exacerbation risk was 8.4 times higher with A. fumigatus sensitization. Multivariate logistic regression revealed higher risk with A. fumigatus sensitization (4.79), fixed obstruction (6.2), systemic steroids use (5.85), indoor mold exposure (4.45), and familial asthma history (2.53). CONCLUSION: A. fumigatus sensitization significantly influences asthma course and control, worsening prognosis. Targeted examination of patients measuring sIgE levels for common aeroallergens is necessary to identify control obstacles. The results of the study were previously reported at the All-Russian Congress on Medical Microbiology, Clinical Mycology and Immunology and published in the form of abstracts in the journal Problems in medical mycology, 2022, Vol. 24, No 2.

Immunological characteristics of airway inflammation in asthma patients with sensitization to various fungal allergens are not well understood, and the search for new markers is necessary to establish future targets for targeted therapy. The purpose of our study was to assess the levels of eosinophilic inflammation markers of the respiratory tract in patients with severe asthma and allergic bronchopulmonary aspergillosis, depending on the spectrum of fungal sensitization. The study included 31 patients with severe asthma with fungal sensitization and 29 patients with allergic bronchopulmonary aspergillosis (ABPA). The levels of total, specific IgE to fungal allergens and periostin in blood serum were determined by enzyme immunoassay. The study of basophil activation was performed by flow cytometry. The patients with severe asthma and sensitization to Aspergillus spp. and Alternaria spp. had significantly higher levels of eosinophils, periostin, and stimulation index to Alternaria spp. in the basophil activation test when compared with the group of asthma patients with sensitization to Aspergillus spp. only. In patients with ABPA with combined sensitization, we have found significantly higher levels of eosinophils, periostin, and stimulation index to Alternaria spp. The finding of pronounced eosinophilic type of inflammation in patients with asthma and combined sensitization to thermotolerant and thermolabile fungi may result from aggressive effect of fungal allergens on the barrier functions of bronchial epithelium, which should be taken into account when choosing therapeutic strategy and administration of immunobiological therapy.

The etiology of precocious puberty includes organic anomalies, genetic mutations, but the primary cause remains unclear in the vast majority of cases. Gonadotropin-releasing hormone (GRH) agonists are used as a treatment of gonadotropin-dependent precocious puberty. Blocking the secretion of gonadotropin-releasing hormone, these drugs stop the premature development of sexual features, prevent premature closure of ossification zones, thereby increasing the child’s expected adult height. The interest in the effects of this group of drugs beyond the hypothalamic-pituitary-gonadal axis has been recently increased. A series of clinical cases have been reported on the development of autoimmune diseases, e.g., autoimmune thyroiditis, Graves disease and type 1 diabetes. The article presents a clinical observation of a patient with central form of premature development who exhibited satisfactory response to treatment with a GRH agonist drug. Further follow-up did not show any reproductive dysfunction. Upon immunological examination, a disturbance was revealed only in the cellular component of immunity. An increased metabolic activity of neutrophils was found, thus, probably, indicating a nonspecific inflammatory process. The levels of immunoglobulins A, M, G matched the reference values. Thus, the therapy with a drug from the group of GRH agonists was effective and safe in terms of influencing the patient’s immune system. The role of hormonal disorders and effects of GRH agonists on the development of immunopathological conditions require further research.

Introduction. Allergy to microscopic fungi (micromycetes), which can colonize the respiratory tract, is associated with an eosinophilic endotype of inflammation and severe disease course. Among the hallmarks of fungal allergy, experts identify fixed airways obstruction, bronchiectasis, and fibrosis. Currently, there has been proposed the term “allergic fungal airways disease” (AFAD), which combines various clinical phenotypes, which pathogenesis is based on IgE-mediated allergic reaction against fungal spores colonizing the respiratory tract. According to this approach, the AFAD group includes severe asthma with fungal sensitization and allergic bronchopulmonary aspergillosis (ABPA). A promising direction in AFAD treatment implies the use of genetically engineered biological drugs against T-2 inflammation. The aim of the study was to evaluate the prevalence ofAspergillusspp. sensitization and allergic bronchopulmonaryAspergillosis in patients with asthma. Materials and methods. A retrospective single-center registry study was conducted in the I.I. Mechnikov North-Western State Medical University, Ministry of Health of Russian Federation. The study included 523 patients with asthma of varying severity. The level of total and specific blood serum IgE against aeroallergens, includingAspergillusspp., was analyzed by enzyme immunoassay. The diagnosis of asthma was established in accordance with the recommendations of the GINA working group and the Clinical recommendations of the Ministry of Health of the Russian Federation, fungal sensitization and ABPA — in accordance with the recommendations of the ISHAM working group.Results. During the examination of patients with asthma, sensitization toAspergillusspp. recorded in 16.6% cases, severe asthma with sensitization toAspergillusspp. — in 7.4%, ABPA — in 6.1%. In patients with allergic asthma, the prevalence of sensitization toAspergillusspp. was 22%, severe asthma with sensitization toAspergillusspp. — 10.1%, ABLA — 8.1%. Among patients with ABPA, females (64%) aged 40 years and older prevailed. The major clinical manifestations of ABPA were presented as cough (96%) and shortness of breath (76%), CT scan radiological changes were in a form of bronchiectasis (72%). Conclusion. A high prevalence of sensitization toAspergillusspp. in patients with asthma was found. It is necessary to includeAspergillusspp. into the list of tested allergens while examining patients with chronic lung diseases for timely detection of AFAD and prescription of proper anti-inflammatory or antimycotic therapy.

Introduction. Aspergillus fumigatus is able not only to sensitize patients with atopy, but also to colonize the respiratory tract, remaining a constant source of allergens due to the small spore size and thermal tolerance. Currently, the role of micromycetes in the immunopathogenesis of asthma has been poorly studied. The objective was to evaluate the features of the immune response regulation in patients with allergic bronchopulmonary aspergillosis (ABPA) or asthma with sensitization to A. fumigatus. Materials and methods. There were enrolled 15 patients with ABPA, 10 patients with asthma with sensitization to A. fumigatus, 16 patients with asthma without sensitization to A. fumigatus. The control group consisted of 16 apparently healthy volunteers. All patients underwent a clinical and functional examination. The subpopulations of blood lymphocytes were assessed by flow cytometry. The A. fumigatus allergen was added to peripheral blood samples to evaluate the production of IFN, IL-10 and IL-13. The serum cytokine levels in cell culture supernatants, as well as total IgE, A. fumigatus-specific IgE (sIgE) as well as thymus and activation-regulated chemokine (TARC) level were measured by enzyme-linked immunosorbent assay. Results. Significantly higher serum levels of total IgE, A. fumigatus-specific sIgE and TARC were found in patients with ABPA and asthma with sensitization to A. fumigatus compared to patients with asthma without sensitization to A. fumigatus. The results of lymphocyte immunophenotyping revealed significant excess of memory Th2 cells and T-regulatory cells in all patients with asthma compared to the control group. The count of Tfh2 was higher but memory Th17.1 cells were lower in patients with sensitization to A. fumigatus compared to those of apparently healthy volunteers. Patients with ABPA had significantly higher count of memory Th2 cells and TARC level compared to patients with asthma sensitized to A. fumigatus. The increased activity of memory Th2 cells is confirmed by increased secretion of IL-13 and IL-10 following along with decreased IFN production in response to specific fungal allergen stimulation of blood cells compared to the patients with asthma and the control group. A positive correlation was revealed between the count of memory Th2 cells and the levels of sIgE, IgE, TARC, a negative correlation with FEV1. Conclusion. Thus, exposure to A. fumigatus significantly enhances the activity of memory Th2 cells in patients with asthma which can lead to severe disease course and development of allergic bronchopulmonary aspergillosis. The features of the immune response identified dictate a need for a personalized approach to choose therapeutic tactics in such patients.

BACKGROUND:Diagnosis of asthma with sensitization toAspergillusspp. is becoming increasingly important due to the severe, uncontrolled course of the disease and the possibility of the formation of allergic bronchopulmonary aspergillosis. AIM:To evaluate the possibility of using the basophil activation test using flow cytometry for the diagnosis of asthma with sensitization toAspergillusspp. MATERIALS AND METHODS:118 patients with asthma were examined. The levels of total IgE and specific IgE to aeroallergens were determined in the blood serum by the enzyme immunoassay. Basophil activation was studied by flow cytometry using the Allergenicity kit (Cellular Analysis of Allergy, Beckman-Coulter, USA). The allergenAspergillus fumigatus(Alcor Bio, Russia) was used to stimulate basophils. RESULTS:The first group consisted of 57 patients with asthma without sensitization toAspergillusspp. The second group included 36 patients with asthma with sensitization toAspergillusspp. The third group consisted of 25 patients with allergic bronchopulmonary aspergillosis. The number of basophils activated by theAspergillusfumigatusallergen in patients with asthma with sensitization toAspergillusspp. and allergic bronchopulmonary aspergillosis was significantly higher than in the asthma group and amounted to 8.1 [5.2; 20.9]% and 84.6 [75.7; 94.0]%, respectively (p 0.001). The stimulation index in the study groups ranged from 0.7 to 72.6. The optimal diagnostic point (cut off) for identifying patients with asthma withAspergillusspp. sensitization there was an stimulation index value of more than 2.4, and for patients with allergic bronchopulmonary aspergillosis 15.95. Among all patients with sensitization toAspergillusspp. a positive correlation was established between the level of specific IgE toAspergillusspp. and the percentage of basophils activated by the allergenAspergillusfumigatus(r= 0.792,p 0.001) and stimulation index (r= 0.796,p 0.05). CONCLUSIONS:The basophil activation test can be used as an additional diagnostic method for asthma with sensitization toAspergillusspp. and allergic bronchopulmonary aspergillosis.

Aim: to assess the clinical and immunological characteristics of patients with severe bronchial asthma (BA) with Aspergillus sensitization and to study the possibilities of applying basophil activation test (BAT) using flow cytometry to detect mycogenic sensitization. Patients and Methods: 93 patients with severe BA were examined. Therapy had 4 or 5 steps according to GINA 2019 recommendations. TARC (thymus and activation-regulated chemokine) content, the number of eosinophils, the levels of total IgE and specific IgE to 10 allergens were determined in blood serum by enzyme immunoassay. BAT with Aspergillus fumigatus allergen was performed in vitro using flow cytometry. Results: Aspergillus sensitization was detected in 33 (35.4%) patients with severe BA. In patients with BA and Aspergillus sensitization, the disease course was uncontrolled, and patients in this group were credibly more likely to receive oral glucocorticosteroids. The number of basophils (activated by the A. fumigatus allergen) and the stimulation index in patients with BA and Aspergillus sensitization were significantly higher than in patients with BA (9.9 (6.0–24.0) % vs. 3.6 (2.0–5.4) %; (p=0.000) and 4.25 (2.49–9.30) vs. 0.94 (0.75–1.16); (p=0.000)). Significant differences in TARC content were obtained in the groups of patients with severe BA and Aspergillus sensitization and patients with BA (625.0 (418.4–875.0) pg/mg versus 406.0 (210.0–561.0) pg/mg; p=0.001). A negative correlation was determined between TARC levels and a decrease in forced expiratory volume in 1 second (FEV1) (r=-0.70, p<0.05), and between positive correlation and absolute eosinophil count (r=0.81, p<0.05) and level of specific IgE to Aspergillus (r=0.36, p<0.05). Conclusion: Aspergillus sensitization is associated with an uncontrolled BA course. An additional method for diagnosing mycogenic sensitization is the BAT. The TARC concentration can serve as a biomarker of an active inflammatory response. KEYWORDS: Aspergillus spp., severe bronchial asthma, mycogenic sensitization, basophil activation test, TARC. FOR CITATION: Kozlova Ya.I., Frolova E.V., Uchevatkina A.E. et al. Clinical and immunological characteristics of severe bronchial asthma with Aspergillus sensitization. Russian Medical Inquiry. 2021;5(1):10–16. DOI: 10.32364/2587-6821-2021-5-1-10-16.

Allergic bronchopulmonary aspergillosis (ABPA) is a severe lung disease caused by hypersensitivity to Aspergillus spp. The search for new diagnostic markers is essential for early detection of the disease in patients at risk and identifying eff ective treatment strategies. 21 patients with ABPA, 37 patients with asthma and 16 healthy subjects were examined. The content of thymic stromal lymphopoietin (TSLP), thymus-associated regulatory chemokine (TARC), IL-8, eosinophil count, total IgE levels and specifi c IgE to Aspergillus fumigatus were determined in serum by ELISA. The positive correlation of the TARC level with the indicators of total IgE, sIgE to Aspergillus fumigatus, the number of eosinophils and the negative correlation relationship with the FEV1 value confi rms the important diagnostic value of this pro-infl ammatory chemokine in patients with ABPA.

Aspergillus fumigatus colonization in the patients with cystic fibrosis (CF) may cause sensitization against A. fumigatus and/or allergic bronchopulmonary aspergillosis (ABPA), which significantly worsens the course of underlying disease. At the present time, new diagnostic tests are searched for detection of fungal sensitization in these patients. The aim of this work was to evaluate an opportunity of application of basophile activation test with A. fumigatus allergen in vitro using flow cytometry, aiming for identification of fungal sensitization in the CF patients. The study included 190 patients with CF aged 1 to 37 years. All the patients underwent common allergy screening (skin tests with fungal allergens, determination of serum levels of total IgE and specific IgE for the fungal allergens), and mycological examination (microscopy and culture of respiratory substrates). Computed tomography of the chest was performed upon clinical indications. The basophil activation test with the A. fumigatus allergen was performed in 10 CF patients with ABPA, and 10 CF patients without ABPA, in addition to the standard allergological examination. Frequency of sensitization to A. fumigatus in the patients with cystic fibrosis was 27%, the incidence of allergic bronchopulmonary aspergillosis was 5.7%. The number of eosinophils, total IgE and specific IgE levels in CF patients with ABPA were significantly higher than in CF patients without ABPA. In blood of the ABPA patients we have identified 68.5 (52.5-81.5%) of basophilic leukocytes activated by A. fumigatus allergen, with a stimulation index of 17.07 (10.30-27.70). In appropriate comparison group, the stimulation index did not exceed 1.5 (p = 0.000). Direct positive correlation between the levels of specific IgE to A. fumigatus and the number of basophils activated by A. fumigatus allergens was revealed (r = 0.77; р < 0.05). FVC values and the body mass index in CF patients with ABPA were significantly lower when compared with the patients without fungal sensitization. Introduction of the basophil activation test, along with standard techniques, may enable a more differentiated assessment of ABPA development in CF patients. Timely detection of associations between A. fumigatus sensitization and clinical status of CF patients will facilitate early and effective administration of specific therapy.