Epigenetics of bronchial asthma
Bronchial asthma (BA) is a chronic inflammatory disease of the respiratory tract. The etiopathogenesis of BA is diverse and is still poorly understood. Epigenetic mechanisms of gene expression regulation are believed to be an important factor associated with the clinical heterogeneity of this disease. A number of studies have established the relationship of individual epigenetic features with predisposition to BA, with the severity of clinical manifestations of the disease, and with the nature of response to drug therapy. In particular, DNA methylation, regulation of genetic activity by microRNA molecules, and modifications of histones are considered to be important mechanisms of epigenetic regulation of inflammatory response of BA. In the review these mechanisms are examined in the context of BA pathogenesis and therapy and information on the main methods for assessing epigenetic modifications is provided. Further study of epigenetic aspects of BA is relevant and important not only from the point of view of basic science, but also of practical medicine, due to the possibility of personalization of BA diagnosis and therapy.
A comparative characterization of macrophages tolerant to NOD1 and TLR4 receptor agonists
Introduction. Innate immune cells activated through a pattern recognition receptor develop a transient state of tolerance, i.e. lack of pro-inflammatory cytokine production upon repeated stimulation of the same receptor. The phenomenon of tolerance can be used to enhance resistance against pathogens. However, data about mechanisms of tolerance are contradictory.
Aim of the study - to compare transcriptomic and metabolic characteristics of human macrophages tolerant to NOD1 and TLR4 receptor agonists upon restimulation with the same agonist and cross-stimulation.
Material and methods. Macrophages were obtained by culturing blood monocytes from healthy donors with granulocyte-macrophage colony-stimulating factor. Macrophages were stimulated for 24 h by a NOD1 or a TLR4 agonist to induce tolerance, whereafter restimulated with the same or the other agonist. Expression of 17 immune-response-related genes was assessed by reverse-transcription real-time PCR, TNF production by ELISA, intensity of glycolysis by real-time monitoring of extracellular medium acidification rate.
Results. Macrophages tolerant to a NOD1 agonist are characterized by a more complete inhibition of the transcriptional response to restimulation with the same agonist, as compared to LPS-tolerated macrophages restimulated with LPS. We did not observe significant cross-tolerizing effects of NOD1 and TLR4 agonists on gene expression. We also show that in macrophages tolerant to NOD1 or TLR4 agonists, there is a complete inhibition of activation-induced enhancement of glycolysis upon restimulation with the same agonist, and partial inhibition upon cross-stimulation.
Conclusion. These data are discussed in light of main theories of immune tolerance (receptor-based, epigenetic).
The relationship of markers of early and late lymphocytes activation with the efficiency of neoadjuvant chemotherapy in triple negative breast cancer patients
Introduction. Determination of reliable predictive and prognostic biomarkers is essential, both for assessing the clinical course of cancer and for the development of adequate treatment methods.
The aim of the study is to determine the relationship between CD3+CD19– and CD3+CD4+-T-cells, CD3–CD19+-B-cells, activated CD25+-, CD4+CD25+-, HLA-DR+-, CD3+HLA-DR+-, CD3-HLA-DR+-cells and CD16+CD56+Perforin+ peripheral blood lymphocytes (PBL) with immediate and long term results of neoadjuvant chemotherapy (NAСT) with paclitaxel and cisplatin in patients with TN BC.
Material and methods. The study included 76 patients aged 26 to 76 years. The observation period was 66.9 months. Using multivariate cytometric analysis, the relationship between the initial (before chemotherapy) amount of CD3+CD19‒, CD3+CD4+, CD3‒CD19+, CD25+, CD4+CD25+, HLA-DR+, CD3+HLA-DR+, CD3-HLA-DR+, CD16+CD56+Perforin+ lymphocytes stage of disease, degree of therapeutic pathomorphosis (TP), disease progression (DP), overall (OS) and relapse-free survival (DFS) of patients was investigated.
Results. An increase in the initial number of CD25+- и CD4+CD25+-lymphocytes was associated with an increase, and HLA-DR+- and CD3-HLA-DR+-cells with a decrease in DFS and OS in patients compared with patients in whom these parameters were reduced. An increase in the initial number of HLA-DR+- and CD3+HLA-DR+-cells was associated with later stages of the disease, and an increase in the percentage of CD3-HLA-DR+-lymphocytes was combined with a low level of TP.
Conclusion. Populations of CD25+- и CD4+CD25+-PBLs may be possible positive predictive factors in patients with TN BC receiving NACT with cisplatin and paclitaxel. It can be assumed that NAH with paclitaxel and cisplatin will be less effective in patients with stage II–III breast cancer who have an «increased» number of HLA-DR+- and CD3-HLA-DR+-lymphocytes before treatment than in patients with a decrease in the initial number of these cells.
Three-month results of vaccination of mono-stationary health workers with the drug «Gam-COVID-Vac»
Introduction. Antibodies are considered as a key immune effectors that provide protection against pathogenic threats. At the same time, the nature and duration of the antibody response to SARS-CoV-2 infection has not been precisely determined.
The aim of the study was to determine the changes in the level of antibodies to SARS-CoV-2 during immunization with the «Gam-COVID-Vac» vaccine.
Material and methods. Observations were carried out with participance of 30 employees of the city clinical hospital No. 1. Medical workers were vaccinated against COVID-19 with the «Gam-COVID-Vac» vaccine according to the standard scheme. The level of antibodies was assessed on the 17th day after the administration of the 1st component of the vaccine, on the 17th and 30th days after the administration of the 2nd component of the vaccine.
Results. Our studies show that the blood level of IgM antibodies to SARS-CoV-2 practically did not change on the 17th and 38th days after immunization with the 1st component (the average CP values were 1.616 and 1.75, respectively). However, by the 30th day, a decrease of IgM to SARS-CoV-2 was observed (average CP value - 0.7829). It should be noted that on the 17th day after immunization a significant variation was revealed in its indicators, which leveled off by the 30th day. A completely different picture was observed with the level of IgG antibodies to SARS-CoV-2. It significantly increased after immunization with the 2nd component and did not change until the 30th day (the average CP values were 12.36 and 12.48 on the 38th and 51st days, respectively). It is necessary to pay attention to the fact that after immunization with the 1st component on the 17th day, a low level of IgG to SARS-CoV-2 (CP - < 1) was recorded in 48 %, and after immunization with the 2nd component - only in 10 % vaccinated. At the same time by 51 days from vaccination the seroconversion level was 100 %. It should be noted that in some patients the IgG level changed by the 30th day after immunization, compared to the 17th day.
Conclusion. Thus, our information confirms that the «Gam-COVID-Vac» vaccine is effective and, in more than 90 % of cases, leads to the formation of a sufficiently high level of antibodies against SARS-CoV-2.
Production of transforming growth factor β and interleukin-10 in the mouse spleen at early stage of gestation in models of spontaneous and muramylpeptide-dependent abortions
Introduction. The maternal immune response is a key determinant of successful pregnancy development. Among the multiple local and systemic mechanisms of immune tolerance formation to the fetoplacental «graft», the regulatory cytokines interleukin-10 (IL-10) and transforming growth factor β (TGFβ) play the leading role. However, the relationship between the production levels of IL-10 and TGFβ in peripheral organs of the immune system in vivo and the frequency of fetal losses in experimental pregnancy failure has not been sufficiently studied.
The aim of the study was to evaluate the production levels of IL-10 and TGFβ in the spleen of pregnant mice in models of spontaneous and muramylpeptide-dependent abortions at early gestational age.
Material and methods. Allogeneic physiological pregnancy was reproduced by crossing CBA/lac females (H-2k) with Balb/c males (H-2d) and the model of spontaneous abortion by mating females with DBA/2J males (H-2d). To model induced and potentiated abortions, CBA/lac females fertilized by Balb/c or DBA/2J males, respectively, were intraperitoneally injected with muramyl dipeptide β-heptylglycoside (C7MDP) at the dose of 20 μg per 1 animal on days 5 and 7 of gestation (DG). Animals were removed from the experiment at the 8th DG. Cytokine-producing cells in spleen sections were detected by immunohistochemical analysis using rabbit polyclonal antibodies to TGFβ and goat polyclonal antibodies to IL-10. The density of TGFβ- and IL-10+ cells was determined by morphometry using Image Scope M software. The digital data were analyzed using Sigma Stat 3.5 software, the differences were considered significant at p < 0.05.
Results. The density of IL-10+- and TGFβ+-cells in the spleen in spontaneous abortions was half that in physiological pregnancy. Under the conditions of baseline high-fertility crosses, injection of C7MDP into pregnant mice (induction of abortion) resulted in an increase in the density of IL-10+-cells, while the density of TGFβ+-cells did not change. In pregnant mice with initially high levels of spontaneous reproductive losses, exposure to C7MDP (abortion potentiation) caused an increase in the density of IL-10+- and TGFβ+-cells. Localization of TGF-β in spleen functional structures did not differ between physiological pregnancy and spontaneous abortions; muramylpeptide-dependent abortions were characterized by an increase in TGFβ-positive structural and functional areas.
Conclusion. The increased incidence of fetal losses compared with physiological pregnancy occurred against the background of decreased IL-10- and TGFβ-producing cell density in the spleen in spontaneous abortions and increased IL-10+-cell density in induced abortions. TGFβ-production was increased in potentiated abortions compared to spontaneous abortions. The increased density of IL-10+-cell distribution in induced abortions and the increased localization of TGF-β+-zones in the spleen in induced and potentiated muramylpeptide-dependent abortions did not abolish the abortogenic effect of C7MDP.
Phenotype and function of neutrophils in premature infants and their role in neonatal infections
Introduction. Premature infants are at high risk bacterial infections development due to functional deficiency of neutrophils.
The aim of the study - to determine Fey-receptors expression (CD64, CD16, CD32) and respiratory burst intensity of neutrophils in preterm infants, as well as to assess their diagnostic value in identifying infectious complications of the early neonatal period.
Material and methods. Premature newborns (n = 102) were divided into 3 groups: neonates without infections in the early neonatal period (n = 24), neonates with localized bacterial infections (n = 63) and early neonatal sepsis (n = 15). Expression of CD64, CD16, CD32 by cord blood neutrophils and intensity of oxidative burst after stimulation of neutrophils with E. coli in the presence of dihydrorhodamine 123 were measured by flow cytofluorometry.
Results. In the group of premature infants who developed early neonatal sepsis a significant increase in CD64 expression, a decrease in CD32 expression as well as a significant increase in the production of reactive oxygen species by neutrophils in response to E. coli were found in comparison with groups of newborns with a localized infectious process and without an infectious diagnosis. The production of reactive oxygen species was also increased in premature infants with a localized infectious process in comparison with the group of premature infants without an infectious diagnosis.
Conclusion. The expression levels of CD64 and CD32 as well as the production of reactive oxygen species by neutrophils of preterm infants, are associated with the development of neonatal sepsis. Determination of these parameters has diagnostic value in identifying infectious complications in early neonatal period.
Immunological mechanisms of the drug Cytovir®-3 action as the basis of the prevention of acute respiratory viral infections and influenza
Introduction. Influenza viruses and other acute respiratory viral infections (ARVI) cause massive outbreaks of infectious diseases quite often becoming epidemic nature. The economic and social damage done to the health of the population by these infections and related complications is enormous. It is quite naturally, many research teams are persistently looking for ways and means of solving this serious problem. The complex preparation Cytovir®-3 in the form of capsules has been used for 20 years for the prevention and early pathogenetic treatment of influenza and ARVI. The study of the mechanism of anti-infectious protection is one of the main tasks in the development of a antiviral medicinal product with both direct and indirect effects.
The article presents the results of an open non-randomized clinical study of the safety and tolerability of the drug Cytovir®-3 capsules in healthy volunteers aged 18 to 30 years with a prolonged course of taking the drug with the study of immunological mechanisms of action
Aim of the study - to investigate the immunological mechanisms of action and safety of Cytovir®-3 capsules in healthy volunteers with a prolonged course of administration.
Material and methods. The article presents the results of an open, non-randomized clinical study of the safety and tolerability of Cytovif®-3 capsules in 21 healthy volunteers aged 18 to 30 years. The duration of the course of taking Cytovir®-3 was 14 days, 1 capsule 3 times a day. An in-depth examination of the volunteers was carried out at the screening stage, on the 5th day of taking the drug and after the 14-day course. Safety and tolerance were assessed according to the data of clinical examination with fixation of vital signs, as well as by studying the dynamics of clinical and biochemical blood tests, general urine analysis and indicators of the body’s immunological reactivity.
Results. In the course of the clinical study the activating effect of the drug on the indicators of stimulated oxidative NBT activity and the level of lysosomal cationic proteins, the phagocytic index, as well as the dynamics of secretory IgA (sIgA) was noted. 4 Cases of adverse events (headache and itching) were registered in 3 (14.3 %) participants, they were regarded as mild and unlikely to be related to the drug intake.
Conclusion. The studied scheme of the prolonged course of taking the drug Cytovir®-3 capsules showed the activity of the drug in relation to the microbicidal parameters of the cells of innate immunity (functional activity of monocytes, neutrophils, as well as the level of sIgA), the fluctuations of which were significant and localized within the limits of the adaptive response norm, which indicates an increase in the body’s immunoresistance after the course and reveals the mechanisms of the preventive action of the drug was shown a high level of safety and good tolerance in assessing the effect on the vital functions of the body, instrumental, laboratory general clinical and immunological indicators.
Immunological aspects of the effects of mesotherapy with hyaluronic acid
Introduction. Despite numerous studies of the immunological aspects of mesotherapy with hyaluronic acid injections, not all of the effects have been studied.
The aim is to study the dynamics of the immunocompetent cells functioning during meso-correction with hyaluronic acid preparations, as well as to determine its effect on the function of the thyroid gland and psychoemotional state.
Material and methods. During dynamic observation of 26 patients before, just after, 3 and 6 months after the course of mesocorrection with hyaluronic acid preparations, the effects on the features of the clinical course, the dynamics of thyroid hormones and the emotional state. «Aramo Smart Lite 300» was used to objectify clinical changes. Quantitative and functional indicators of the innate and adaptive immune response were determined to characterize the immune system.
Results. Improvement in the parameters of skin moisture, the relief of skin creases was revealed, the tendency to decrease was determined only six months after the course of correction. The results of the analysis of immunological parameters indicate dynamic changes marked at the end of the course in the form of lymphocyte differentiation shift towards NK-cells and B-lymphocytes, increased production of IgM and IgG, decreased levels of IgA and IgE in serum, inhibition of oxygen-producing activity of neutrophils, changes of Toll-like receptors expression by monocytes. Further observation showed a return of all indicators to the initial level. The study of the quality of life of patients showed a significant improvement in the psychoemotional state immediately after the course and the persistence of improvements throughout the follow-up period.
Conclusion. The clinical effect is accompanied by changes in the parameters of the innate and adaptive immune response, recorded immediately after the completion of the procedure, which serves as an illustration of the involvement of immune processes in improving the moisture content, elasticity and relief of the skin.
Hybrid proteins CFP10 and ESAT6. A path from developing a molecule to population screening for TB infection
The review presents the history of the creation and mass production of Diaskintest® - a drug for mass screening of the population to detect a tuberculosis infection. The molecule contains two proteins ESAT6 and CFP10 that are only found in virulent Mycobacterium tuberculosis and are absent in the M. bovis BCG vaccine strain. The prognostic value of the drug is determined by the fact that these proteins are secreted by dividing Mycobacterium tuberculosis only, and not by that in a dormant state. Such a genetically engineered product required the creation of high-tech manufacturing facilities for mass production of the drug that would meet the needs of the entire country, which was done in the shortest possible time. This diagnosticum is intended for intracutaneous administration and its administration technique is similar to the Mantoux test, which made the test available for mass use. In preclinical studies, the skin test using Diaskintest® has proved its safety, high specificity and sensitivity. Clinical studies the results of which have been confirmed by broad clinical practice, as well as post-marketing surveillance demonstrated that the test has high specificity, sensitivity, and prognostic value. Mass screening of the child and adolescent population resulted in a sharp decrease in morbidity and an improvement in the structure of clinical forms in patients. Using the test in risk groups in adults makes it possible to identify persons with tuberculosis and those in need of preventive treatment. The Diaskintest® test has proven its effectiveness in clinical practice when used in primary health care.