Бронхиальная астма в сочетании с полипозным риносинуситом: клиническая характеристика и анализ локальной экспрессии гена IL371 Федеральное государственное бюджетное учреждение «Государственный научный центр «Институт иммунологии» Федерального медико-

Background: Mouse models of allergic asthma play a crucial role in exploring of asthma pathogenesis and testing of novel anti-inflammatory drugs. Widely used acute asthma models usually developed with adjuvant (aluminum hydroxide (alum)) do not reproduce one of the main asthma feature - airway remodeling while chronic asthma model mimic the pathophysiology of human disease. Moreover, the use of alum causes distress in experimental animals and impedes the test of adjuvant-containing drugs. In this study, we aimed to develop a chronic adjuvant-free asthma model with pronounced asthmatic phenotype. Methods: Female BALB/c mice were divided into 3 groups. The first group was sensitized with intraperitoneal injections of ovalbumin (OVA) emulsified in aluminum hydroxide on days 0, 14, 28 followed by two stages of intranasally challenge with OVA on days 41-43 and 62-64. The second group was subcutaneously sensitized with the same dose of OVA without adjuvant and challenged on the same days. The third group (negative control) included mice which did not received any kind of treatment (i.e. sensitization and challenge). Serum levels of OVA-specific IgE, IgG2a and IgG1 antibodies were detected by ELISA. Airway hyper-responsiveness was measured by non-invasive plethysmography on days 44 and 65. Bronchoalveolar lavage fluids (BALF) sampled in all groups on days 45 and 66 were analyzed by light microscopy. The left lung was removed for histological analysis. The IL-4 and IFN gamma mRNA expression in BALF cells was evaluated by RT-PCR. Results: The OVA-specific IgE antibody response was two-fold increased in mice from adjuvant-free group compared to the adjuvant group that reflects reorientation of immune response towards Th2 phenotype. At the same time, the level of OVA-specific IgG1 and IgG2a antibodies was increased in the adjuvant group. Airway hyperresponsiveness to methacholine in mice of both experimental groups was two-fold higher than in control. Analysis of cell composition in BAL has shown a significant increase in eosinophil count in both experimental groups that indicate the development of allergic inflammation. Lung histology revealed airway remodeling in both experimental groups including goblet cell hyperplasia/metaplasia, thickening of airway walls, collagen deposition in the wall of distal airways. Additionally, the tendency to develop hypertrophy of bronchial smooth muscle layer was observed. Study of gene expression in BAL cells revealed the increase of IL-4 level in both adjuvant and adjuvant-free groups while IFN gamma expression in both experimental groups was similar to control group. Conclusion: We have developed a chronic adjuvant-free mouse asthma model which possesses all necessary features of the disease including airway remodeling and is more suitable for pre-clinical evaluation of novel therapeutic approaches including adjuvant-containing drugs.

Interleukin-33 (IL-33) belongs to the IL-1 cytokine family and plays an important role in modulating immune system by inducing Th2 immune response via the ST2 membrane receptor. Epithelial cells are the major producers of IL-33. However, IL-33 is also secreted by other cells, e.g., bone marrow cells, dendritic cells, macrophages, and mast cells. IL-33 targets a broad range of cell types bearing the ST2 surface receptor. Many ST2-positive cells, such as Th2 cells, mast cells, basophils, and eosinophils, are involved in the development of allergic bronchial asthma (BA). This suggests that IL-33 directly participates in BA pathogenesis. Currently, the role of IL-33 in pathogenesis of inflammatory disorders, including BA, has been extensively investigated using clinical samples collected from patients, as well as asthma animal models. In particular, numerous studies on blocking IL-33 and its receptor by monoclonal antibodies in asthma mouse model have been performed over the last several years; IL-33-and ST2-deficient transgenic mice have also been generated. In this review, we summarized and analyzed the data on the role of IL-33 in BA pathogenesis and the prospects for creating new treatments for BA.

In the study, the effect of the TLR4 agonist Immunomax was investigated in vitro and in vivo. In particular, Immunomax was shown to polarize mouse bone marrow macrophages from the M0 and M2 states into the M1 state (ARG1 and iNOS mRNA expression levels were used to identify the mouse M1 and M2 phenotypes). Next, we investigated the prophylactic antiviral effect of Immunomax in both a model of mouse respiratory syncytial virus (RSV) infection and a model of RSV-induced bronchial asthma (BA) exacerbation. In the experiment with RSV-induced BA exacerbation, Immunomax-treated mice were characterized by a significant decrease of the viral load in lung homogenates, an increased amount of M1 macrophages in the lung, a tendency toward Th2-dependent ovalbumin-specific IgG1 antibodies decrease in blood serum, a significant increase in RSV-activated CD4+ T cells secreting IFNγ (Th1 cells), and a simultaneous significant decrease in the amount of CD4+ cells secreting IL-4 (Th2 cells) in the mouse spleen, which were detected by ELISPOT 1.5 months after experiment. These findings suggest that treatment with the TLR4 agonist Immunomax polarizes the immune response towards antiviral Th1 and may be used for short-term antiviral prophylaxis to prevent acute respiratory viral infections in asthmatics.

Bronchial asthma (BA) is a heterogeneous disease which characterized by chronic inflammation of the respiratory tract. Respiratory viruses such as the respiratory syncytial virus (RSV) are the main causes of BA exacerbations. Increase of BA incidence stimulates the search for novel targets, which will be suitable for the drug development. Different studies have shown that IL-33 is involved in the pathogenesis of bronchial asthma, but there is no enough information on its role in the pathogenesis of virus-induced of BA exacerbations. Therefore, the purpose of this study was to evaluate the effect of suppression of the il-33 gene by small interfering RNA (siRNA) molecules on the mouse model of RSV-induced BA exacerbations. The local suppression of il-33 gene in lung tissue was shown to decrease the main features of the disease: the pulmonary function was restored, and the degree of pro-inflammatory cells in lungs was reduced. Simultaneously, suppression of il-33 did not alter RSV-infection of respiratory tract. IL-33 was proved to be a promising target for the development of new drugs for the therapy of bronchial asthma and its virus-induced exacerbations.

Asthma is among the most common chronic disorders of airways, which affects both children and adults. Asthma being a common disease among different segments of population, it has a high mortality rate and, in the absence of appropriate care, affects the quality of life and leads to economics losses. In a view of continuing growth in the incidence of asthma, it is important to find relevant biological targets for developing new approaches to astma therapy. Recent advances in molecular immunology, genetics, and bioinformatics allowed genes involved in the pathogenesis of asthma to be identified, which provided prerequisites for the development of new types of drugs that can regulate the activity of pathogenically significant genes. To date, a number of technologies for sequence-specific gene regulation (ASO, ribozymes, DNAzymes, EGS, DNA-decoys, U 1-adapters) are available, but RNA interference is the most promising approach in both terms of efficacy and financial cost. This review focuses on the generalization and analysis of experimental data regarding the use of RNA interference technology for the treatment of astma.

Bronchial asthma (BA) is the most common chronic disease of the airways affecting up to 18% of population in different regions of the world. Respiratory viruses such as rhinoviruses and respiratory syncytial virus (RSV) are the main causes of BA exacerbations. Current data demonstrate the major role of Th1- and Th2-response and corresponding cytokines in the development of infectious and allergic inflammation of the airways. At the same time, there are no data available concerning changes in Th1/Th2-balance during virus-induced asthma exacerbations. In the current study, we evaluated Th1/Th2-balance using developed mouse model of RSV-induced BA exacerbation. In was demonstrated, that RSV infection does not increase Th2-immune response but enhances Th1-response that shifts Th1/Th2-balance towards Th1.

Бронхиальная астма является одним из самых распространенных хронических воспалительных заболеваний дыхательных путей, от которого страдают как взрослые, так и дети. Данное заболевание распространено среди разных слоев населения, имеет высокий уровень смертности, а при отсутствии квалифицированной помощи приводит к ухудшению качества жизни населения и экономическим потерям для государства. Ввиду продолжающегося роста заболеваемости бронхиальной астмой остается актуальной задача поиска значимых биологических мишеней с целью разработки новых способов терапии. Недавние успехи в области молекулярной иммунологии, генетики и биоинформатики позволили выявить гены, вовлеченные в патогенез бронхиальной астмы, что создало предпосылки для появления лекарственных препаратов нового типа, способных регулировать активность патогенетически значимых генов. К настоящему моменту известен ряд технологий для сиквенс-специфической регуляции генов (АСО, Рибозимы, ДНКзимы, EGS, ДНК-ловушки, U1-адапторы), однако использование феномена РНК-интерференции является наиболее перспективным с точки зрения эффективности и экономической целесообразности. Данный обзор посвящен обобщению и анализу экспериментальных данных по использованию технологии РНК-интерференции для терапии бронхиальной астмы.

Background : Aluminum hydroxide (alum) is widely used for sensitization in mouse models of allergic asthma (MMA). The purpose of this study was to develop a phenotype of allergic airway inflammation in adjuvant-free MMA. Methods: BALB/c mice were sensitized according to two protocols. Adjuvant protocol (AP) consisted of three i.p. injections of 20 μg ovalbumin (OVA) with 2 mg of alum/mouse on days 0, 14 and 28 followed by five intranasal applications (INA) with 50 μl OVA (10 mg/ml) on days 41-45. Adjuvant-free protocol (AFP) included by three s.c. injections of the same OVA dose followed by INA with OVA at the same manner. 24 hours after the last INA airway hyperresponsiveness (AHR) to methacholine was measured by plethysmography. 48 hours after the last INA bronchoalveolar lavage (BAL) was performed for differential cell counts and lungs were removed for histological examination. OVA-specific serum IgE-, IgG1- and IgG2a- antibodies were detected by ELISA. Results: OVA-specific IgE production in AFP group was 1.6 fold higher than that of AP group. OVA-specific IgG2a level in AFP group was 3 times lower than that of AP group while levels of anti-OVA IgG1 were similar in both groups. AHR in AP and AFP groups was higher than that of normal mice but did not differ from each other. Both AP and AFP groups have an equal amount of inflammatory cells in BAL while lung histology picture (perivascular and peribronchial inflammation) was more expressive in AFP group. Conclusion: Adjuvant-free protocol generates an experimental asthma phenotype similar to that of adjuvant protocol which is widely used in the literature. The adjuvant-free MMA can be a relevant model for testing of new approaches in treatment or prevention of bronchial asthma.