A nonspecific component of BCG vaccination

Decades of research on the mechanisms of immunological protection against Mycobacterium tuberculosis, the causative agent of tuberculosis (ТВ), did not allow us to draw a final conclusion about the relative importance of specific pathways when forming protective immunological memory. The BCG vaccine, being the only so far approved tuberculosis vaccine protects children from severe forms of ТВ infection. It is still unclear why BCG does not save from primary infection, reactivation of ТВ and latent carrying. At the same time, the association between BCG vaccination and a reduced risk of non-mycobacterial infections, allergies, cancer and general mortality has been demonstrated. Such nonspecific effects of BCG are dependent mostly on the innate immune cells, rather than on specific memory Т cells. There is evidence of an adjuvant effect of BCG vaccination with respect to the humoral immune response to a variety of childhood vaccines. This review is focused mostly on the analysis of works aimed at studying the relatively recently identified mechanism for generating the non-specific effect of the BCG vaccine, i.e., development of induced natural immunity. This phenomenon is mediated by NOD2 signaling and epigenetic macrophage modification and, due to BCG vaccination, leads to enhanced capacity of macrophages to produce TNFa and IL-6 in response to stimulation by BCG-nonrelated microorganisms or TLR ligands. Induced immunity does not only reshape transmission of immunological signals between the cells of innate immune system, but also induces profound changes in the balance of metabolic pathways, such as glycolysis, oxidative phosphorylation, metabolism of amino acids and fatty acids, being accompanied by enhanced ability of innate immune cells to respond to the secondary stimulation. Realizing these intracellular processes opens up new opportunities for therapeutic intervention into the regulation of immune processes accompanying infectious and inflammatory diseases. In general, an in-depth study of the non-specific component of BCG vaccination should lead to emergence of new ideas about the mechanisms of its protective action, as well as affect development of a new ТВ vaccine. This knowledge can stimulate changes in global vaccination policy, aiming for optimization of vaccination benefits and reduction of childhood morbidity and mortality, as well as decrease of the post-vaccinal complications

Effect of tobacco smoke and nicotine on immune response in tuberculosis infection and other lung diseases

The problem of smoking, as well as incidence of tuberculosis, has existed for a long time. The latest WHO data indicate that 1.3 million people die from tuberculosis, and another 7 millions die from smoking every year. Tobacco smoke contains many harmful chemicals, including carbon monoxide, nicotine, nitrogen oxides, and cadmium. A number of studies indicate a high prevalence of smoking among patients with tuberculosis. In most cases, infection with Mycobacterium tuberculosis does not lead to active disease, due to the development of a balanced, homeostatic immune response. The key protective components are inflammatory responses aimed at inhibition of the pathogen growth, its sequestration and final elimination. At the same time, excessive or inadequate immune response may lead to granuloma destruction, tissue damage and, as a result, prolonged duration of treatment due to decreased respiratory function of the lungs. Along with pro-inflammatory mediators, anti-inflammatory mediators are synthesized in the host organism, which can positively or negatively affect the course of disease, depending on the place and time of their production. The balance between pro-and anti-inflammatory mediators in terms of time and expression level plays a crucial role in determining the outcome of infection. In our review, we consider the impact of tobacco smoke on various components of the human immune system, as well as upon the course and outcome of tuberculosis and other lung diseases. In addition, we would like to draw the reader’s attention to the need of adjusting pathogenetic therapy of bronchopulmonary diseases, taking into account the patient’s smoking habits. Tobacco smoking is one of the main causes of the severe course of many infectious and non-infectious diseases of the bronchopulmonary system. The decay products of cigarette smoke disrupt the functioning of the ciliated epithelium of respiratory tract, the production of the mucous component in the bronchi, and reduce the effectiveness of the surfactant system. These negative events interfere with protective mechanisms of the human respiratory system. It is worth of note that tobacco smoke also exerts a systemic effect on the immune system. Data are accumulating on the association between the terms of exposure to tobacco smoke, and a range of adverse tunerculosis manifestations, such as extent of infection, severity of course, reactivation, treatment outcome, and mortality. At the same time, epidemiological studies are able to reveal the associations, but they do not allow us to determine exact causal relationships

EFFECTIVENESS OF COMBINATION OF ISONIAZID AND SILVER NANOPARTICLES IN THE TREATMENT OF EXPERIMENTAL TUBERCULOSIS

Goal of the study: to investigate the impact  of isoniazid combined  with  silver nanoparticles on the isoniazid resistant strains  of tuberculous mycobacteria  and the course of experimental of tuberculosis  caused by the above strains.Materials and methods. The electrochemical metal resolution was used to obtain silver nanoparticles, the average size of nanoparticles made 3-60 nm. The investigated concentrations of silver nanoparticles made 5; 25; 50 µg/ml. The isoniazid was used only in one concentration of 1 mg/ml. Totally there were 651 in vitro tests.The experimental tuberculosis  model included infecting mice (totally 68) with two-week virulent  multiple drug resistant culture  of M. tuberculosis.Results. In vitro tests  proved that  isoniazid combined  with silver nanoparticles fully or significantly  suppressed  the growth  of MDR  TB strain in 49.2% of cases. The minimal inhibitory concentration of nanoparticles in the combination with isoniazid made 2.5 µg/ml and minimal bactericidal concentration made 5 µg/ml. Atomic force microscopy detected the changes in morphometric parameters of MDR  tuberculous mycobacteria after exposure to silver nanoparticles combined with isoniazid unlike the use of isoniazid only or silver nanoparticles. When treating experimental tuberculosis, survival rates and histological tests of the lung tissue confirmed that the combination of isoniazid and silver nanoparticles was preferable compared to the single use of the above components

Saposin D acting on macrophage bacteriostatic function in experimental tuberculosis infection

The protection against tuberculosis infection is largely determined by the ability of host tissue macrophages to limit the growth and spread of mycobacteria. Able to multiply within the host macrophages, mycobacteria have developed a number of protective mechanisms preventing phagosome-lysosome fusion, thereby evading damaging effects of lysosomal enzymes. Saposins are small, acidic, thermostable, non-enzymatic glycoproteins that participate as co-fac-tors in degradation of short oligosaccharide head group glycosphingolipids. Saposins A, B, C and D are formed in acidic endosomes due to cleavage of initial prosaposin molecule. The effect of saposins on human immune response is mediated by their involvement in presenting mycobacterial antigens on CD1 molecules. Preliminary studies with electron microscopy allowed to uncover saposin D-bound damaging effect on Mycobacterium tuberculosis in acidic environment. These data allowed us to suggest that saposin D is an important protective component fighting against TB infection. The aim of the study was to explore how saposin D deficiency might affect formation of anti-tuberculosis immune response and ability of macrophages to inhibit M. tuberculosis growth. Materials and methods. Interstitial pulmonary macrophages and peritoneal macrophages were isolated from wild type C57BL/6 strain and saposin D deficient C57BL/6-SapD-/- mouse strains. Results. It was found that as compared to macrophages from mice, macrophages from wild type strain significantly better controlled mycobacteria growth in vitro. To study an opportunity of compensating for deficient saposin D in peritoneal macrophages from C57BL/6-SapD-/- mice, a saposin D gene-bearing lentiviral vector was created. Transfection of SAPD-deficient peritoneal macrophages with expression vector compensated for saposin D deficiency in such cells and restored bactericidal function. The mechanisms of action for current anti-TB drugs are mediated by various metabolic pathways in mycobacteria (inhibited biosynthesis of fatty acids, arabinogalactan, peptidoglycan and protein; inhibition of DNA-dependent processes, proton pumps and cytochrome P450-dependent monooxygenases). Conclusion. It was shown that saposin D deficiency affects activation of macrophage bactericidal function in vitro. Our study data may be a prerequisite for biologically substantiated potential of using a vector construct bearing natural human protein gene such as saposin D, as a new anti-tuberculosis drug

MULTIPLE DRUG RESISTANCE PROTEINS OF PULMONARY SOMATIC CELLS AND THEIR SPECIFIC EXPRESSION IN FIBROUS CAVERNOUS TUBERCULOSIS

Cells of the host containing special proteins-transporters, so-called proteins of multiple drug resistance (MDR proteins) can contribute to the drug resistance formation.Goal of the study: to define specific expression of gene and distribution of main MDR proteins (MDR1/Pgp, MRP1, lRP, BCRP) in the pulmonary cells in case of active tuberculosis.Materials and methods. Expression of MDR protein genes was evaluated by RT-PCR for mRNA isolated from the surgical specimen of fibrous cavernous tuberculosis patients. Localization of MDR proteins was performed by immunohistochemical staining and confocal laser microscopy. Main results. Intensity of MDR protein genes expression varies in different zones of tuberculous lesions: MDR1 and BCRP are characterized by the highest level and MRP1 gene is characterized by the minimum level. The level of lRP gene expression depends on the inflammation zone and it is maximum in perifocal zone where protein is detected in the cells of alveolar epithelium and macrophages. High expression of MDR protein genes in various parts of tuberculous lesions witnesses about the potential involvement of these proteins in the development of drug resistance to anti-tuberculosis drugs

MYCOBACTERIOSIS IN THE PRACTICE OF PULMONOLOGISTS AND PHTHISIOLOGISTS

Goal of the study: to investigate the frequency of mycobacteriosis in the practice of pulmonologists and phthisiologists.Materials and methods. The analysis included medical files of 156 patients with various non-specific respiratory diseases and tuberculosis who were examined in 2011-2014 and non-tuberculous mycobacteria were found in their sputum.Results of the study. pulmonary mycobacteriosis was diagnosed in 93 (59.6%) patients out of 156 who corresponded the criteria of American Thoracic Society and etiotropic treatment was prescribed. While detection of non-tuberculous mycobacteria in the other patients was considered as innidiation of respiratory tract by those mycobacteria in case of chronic non-specific pulmonary disease and/or tuberculosis. Mycobacterioses were treated with the relevance to drug susceptibility of the causative agent and there were numerous obstacles due to high frequency of adverse reactions

Analysis of drug resistance indicators of Mycobacterium tuberculosis in the Republic of Mary El for 2007-2017

The complex analysis of drug resistance indicators of Mycobacterium tuberculosis to anti-tuberculosis drugs for different categories of tuberculosis patients in the Republic of Mari El for the period from 2007 to 2017 was carried out. There is a significant spread of drug-resistant strains of Mycobacterium tuberculosis even among the new cases of tuberculosis. There is a steady increase in the level of primary and acquired drug resistance, especially multi and extensively drug resistance. The presence in the Republic of Mari El of the bacillary nucleus of chronic forms of tuber-culosis with a wide range of resistance was revealed, which creates prerequisites for the growth of primary drug resistance. To improve the epidemiological situation in the region, it is necessary to op-timize the algorithm of microbiological diagnosis of tuberculosis with the inclusion of methods that ensure the rapid testing of drug resistance to the maximum possible range of anti-tuberculosis drugs. This will make it possible to prescribe adequate treatment in a timely manner and prevent the further development of resistance and the spread of drug-resistant strains of Mycobacterium tuberculosis.Проведен комплексный анализ показателей лекарственной устойчивости микобактерий туберкулёза к противотуберкулезным препаратам для различных категорий больных туберкулёзом в республике Марий Эл за период с 2007 по 2017 гг. Установлено, что даже среди впервые выявленных больных туберкулёзом имеет место значительное распространение лекарственно-устойчивых штаммов. отмечено неуклонное нарастание уровня первичной и приобретенной лекарственной устойчивости, особенно множественной и широкой. Выявлено наличие в республике Марий Эл бациллярного ядра хронических форм туберкулёза с широким спектром резистентности, что создаёт предпосылки для роста первичной лекарственной устойчивости. для улучшения эпидемиологической ситуации в регионе необходима оптимизация алгоритма микробиологической диагностики туберкулёза с включением в него методов, обеспечивающих быстрое тестирование лекарственной устойчивости к максимально возможному спектру противотуберкулезных препаратов. Это позволит своевременно назначать адекватное лечение и предотвращать дальнейшее развитие резистентности и распространение лекарственно-устойчивых штаммов микобактерий туберкулёза

Легочная гипертензия и возможности ее коррекции у больных с тяжелой формой бронхиальной астмы

The study was aimed at the analysis of a possibility of the lung hypertension correction for patients suffering from a severe bronchial asthma with due consideration to various treatment.48 patients with bronchial asthma were examined. Four clinical groups were specified. The first group (14 patients) took angiotenzin-transforming enzyme inhibitor (iATE) — enalapril with an average dose of 20 mg/day.The second group (14 patients) took enalapri in combination with the plasmapheresis. The third group (8 people) took plasmapheresis only. The fourths group (12 patients) was a control group. These patients took basic therapy preparations only. These preparations were used for treating patients of all clinical groups.The basic therapy preparations were cholinolytics, methylxanthines, β-agonists and corticosteroids (by indication). The clinical parameters (number of asphyxia attacks and/or coughing, dyspnea, rales in lungs - cumulative indices), indices of the external respiration function and diastolic pressure in the lung artery were assessed weekly. The latter parameter was determined by the impulse Doppler echocardiography technique on the basis of the flow shape at the iung artery’s vaive. The 6-stage Ali-Sadec-Ali scale (1988) was used for the assessment of diastolic pressure in the iung artery.Initially all the patients had high diastolic pressure in lung artery (31 ±2,1) Mercury and low FEV1(49,4±5,1).After the treatment a reliable diastolic pressure decrease together with the FEV1 increase was marked for the patients of the first and second groups. The breathing insufficiency has decreased aiso. These first two parameters did not chang for the third and fourths groups however clinical indices authentically improved.Taking into consideration the results of the fulfilled analysis it is possible to draw a conclusion that in order to reduce the diastolic pressure in lung artery when treating severe bronchial asthma complicated with the lung hypertension, it is possible to use the isolated enalapril treatment or its combination with the plasmapheresis.Цель работы: изучить возможности коррекции легочной гипертензии у больных с тяжелой формой бронхиальной астмы (БА) в условиях разных режимов лечения.Обследовано 48 больных БА. Выделено 4 клинические группы: 1 группа (14 человек) получали ингибитор ангиотензинпревращающего фермента (иАПФ) эналаприл в средней дозе 20 мг/сутки; 2 группа (14 человек) получали эналаприл в сочетании с плазмаферезом (ПА), 3 группа (8 человек) ПА; 4 контрольная группа (12 человек) получали только средства базисной терапии, которые использовались также при лечении больных всех клинических групп.Средства базисной терапии включали в себя холинолитики, метилксантины, β-агонисты и кортикостероиды (по показаниям). Еженедельно оценивались клинические параметры (количество приступов удушья и/или кашель, одышка, наличие хрипов в легких — кумулятивный индекс), показатели ФВД и диастолическое давление в легочной артерии (ДЛА). ДЛА определяли методом импульсной допплерэхокардиографии (ДЭхоКГ) по форме потока на клапане легочной артерии. В оценке ДЛА использовалась 6-ступенчатая шкала Али-Садек-Али (1988).Исходно все больные имели повышенное ДЛА (31 ±2,1) мм.рт.ст. и сниженный ОФВ1 (49,4±5,1). После лечения в 1 -й и 2-й группе отмечено достоверное снижение ДЛА, повышение ОВФ1 , а так же уменьшение выраженности дыхательной недостаточности. В 3-й и 4-й группах ДЛА и ОФВ1 не изменились, а клинические показатели достоверно улучшились.Исходя из результатов проведенного исследования можно сделать вывод о том, что изолированное применение эналаприла или сочетание его с плазмаферезом может быть использовано для снижения уровня ДЛА при лечении больных с тяжелой формой БА, осложненной легочной гипертензией

Трудности ведения больного туберкулезом легких с множественной лекарственной устойчивостью возбудителя и сопутствующей патологией

The article describes the clinical case of the successful cure of disseminated pulmonary tuberculosis (caseous pneumonia) with multiple drug resistance in the patients with several concurrent diseases: COPD, pulmonary emphysema, respiratory failure of the 1st phase, hypertension of the 2nd phase, coronary heart disease, cardiosclerosis, drug-induced hepatitis. Tuberculosis chemotherapy was accompanied by the intensive hepatotrophic, cardiac and urinative therapy with potassium-sparing diuretics. When drug-induced hepatitis developed the extracorporal methods namely discrete machine plasmapheresis were used.Приведено клиническое наблюдение успешного лечения распространенного туберкулеза легких (казеозной пневмонии) с множественной лекарственной устойчивостью у пациента с несколькими сопутствующими заболеваниями: ХОБЛ, эмфизема легких, дыхательная недостаточность I ст, гипертоническая болезнь IIст, ишемическая болезнь сердца, кардиосклероз, токсический гепатит. Химиотерапия туберкулеза сопровождалась усиленной гепатотропной, кардиальной и мочегонной терапией калийсберегающими диуретиками. При развитии токсического гепатита использовались экстракорпоральные методы лечения в виде дискреторного аппаратного плазмафереза

ЭФФЕКТИВНОСТЬ СОЧЕТАНИЯ ИЗОНИАЗИДА И НАНОЧАСТИЦ СЕРЕБРА В ЛЕЧЕНИИ ЭКСПЕРИМЕНТАЛЬНОГО ТУБЕРКУЛЕЗА

Goal of the study: to investigate the impact  of isoniazid combined  with  silver nanoparticles on the isoniazid resistant strains  of tuberculous mycobacteria  and the course of experimental of tuberculosis  caused by the above strains.Materials and methods. The electrochemical metal resolution was used to obtain silver nanoparticles, the average size of nanoparticles made 3-60 nm. The investigated concentrations of silver nanoparticles made 5; 25; 50 µg/ml. The isoniazid was used only in one concentration of 1 mg/ml. Totally there were 651 in vitro tests.The experimental tuberculosis  model included infecting mice (totally 68) with two-week virulent  multiple drug resistant culture  of M. tuberculosis.Results. In vitro tests  proved that  isoniazid combined  with silver nanoparticles fully or significantly  suppressed  the growth  of MDR  TB strain in 49.2% of cases. The minimal inhibitory concentration of nanoparticles in the combination with isoniazid made 2.5 µg/ml and minimal bactericidal concentration made 5 µg/ml. Atomic force microscopy detected the changes in morphometric parameters of MDR  tuberculous mycobacteria after exposure to silver nanoparticles combined with isoniazid unlike the use of isoniazid only or silver nanoparticles. When treating experimental tuberculosis, survival rates and histological tests of the lung tissue confirmed that the combination of isoniazid and silver nanoparticles was preferable compared to the single use of the above components.Цель исследования: изучить влияние изониазида в сочетании с наночастицами серебра на устойчивые к изониазиду штаммы микобактерий туберкулеза (МБТ) и течение экспериментального туберкулеза, вызванного этими штаммами.Материалы и методы. Наночастицы серебра получали  методом электрохимического растворения металла, средний размер наночастиц 3-60 нм. Изучаемые концентрации наночастиц серебра составили 5; 25; 50 мкг/мл. Изониазид использовали только в концентрации 1 мг/мл. Всего было проведено 651 исследование in vitro.Экспериментальную модель туберкулеза  создавали  путем заражения мышей (всего 68 мышей)  двухнедельной  вирулентной культурой M. tuberculosis с множественной лекарственной устойчивостью (МЛУ).Результаты. Как показали исследования in vitro, сочетание изониазида с наночастицами серебра обеспечило полное и значительное подавление роста штамма МЛУ МБТ в 49,2% наблюдений. Минимальная подавляющая концентрация наночастиц в составе нанокомпозита с изониазидом составила 2,5 мкг/мл, минимальная бактерицидная – 5 мкг/мл. Атомно-силовая  микроскопия выявила изменения морфометрических параметров  МЛУ  МБТ  после воздействия наночастиц  серебра в сочетании с изониазидом  в отличие от изолированного использования изониазида  или наночастиц серебра. При лечении экспериментального туберкулеза по выживаемости и гистологической  оценке легочной ткани было подтверждено преимущество сочетания изониазида  с наночастицами серебра перед раздельным использованием компонентов