In vitro genotoxic effects of prooxidant therapy

Chronic obstructive pulmonary disease (COPD) is a major problem in industrialized countries. Much evidence suggest that oxidative stress has been associated with chronic diseases, including COPD. There are a number of good experimental and clinical studies clearly showing a strong relationship between oxidative stress and DNA-damage in several diseases. The present study was aimed at establishing whether genome damage is also exists in COPD. Prooxidant therapy mediates its action through the development of oxidative stress that can lead to oxidative modification of DNA. The role of ozone in the genome damage has received little attention. The genotoxicity of ozone is of interest because ozone therapy is used for the treatment of various chronic conditions, including COPD and may impose a possible risk of genotoxic effects for patients. The purpose of this study was to investigate genotoxic effect of different dosing schedules of prooxidant therapy in vitro on the blood of patients with COPD. Peripheral blood was obtained from 20 COPD patients and 15 age- and sex-matched controls. Our present results have shown that ozone induces DNA-damage in human blood cells in vitro. The severity of oxidative DNA-damage in COPD patients directly depends on the concentration of ozone. Clinical results have shown that DNA-damage is an important part of the pathogenesis of the chronic inflammatory process in the bronchopulmonary system

The effect of environmental factors on the immune-metabolic status of people living in industrial areas of Primorsky region

The estimation of the total effect of the environment on immune-metabolic parameters of people living in industrial areas of Primorsky region was conducted. The study involved 1128 healthy people living in the Primorsky region for at least 10 years. Integrated exposure index (HE) that takes into account the gradient of "response of the body" to the combined effects of multiple environmental factors (climatic, technological, socioeconomic, etc.) was developed. With use of gradient approach in Primorye 4 categories of areas were identified: the relatively favorable (IIE > 0,4), moderate (IIE = 0,3-0,4), relatively unfavorable (IIE = 0,2-0,3) and unfavorable environment (IIE 0,4). With the use of cluster analysis the classification of objects method К-means was performed, which revealed three immune-metabolic phenotypes. Compensated phenotype corresponds to the first phase of the adaptation of the organism - the activation parameters. Increased exposure leads to the formation of subcompensated phenotype, and the destruction of the body of the adaptation fund - to the formation of decompensated phenotype. The distribution of the proportion of healthy individuals with selected phenotypes in the population living in the areas with different environmental pressure shows the following dependencies: compensated and subcompensated phenotypes prevalent in areas with better living conditions, the percentage of decompensated phenotype increased environmental degradation. In areas where environmental load exceeds the capability to adapt the body subcompensated and decompensated phenotypes prevail. Violations identified unfavorable combination of endogenous and exogenous risk factors may be the basis for the formation of disease

The formation of oxidative disorders in the population of Vladivostok under the influence of atmospheric microparticles

We studied the response of trigger systems in healthy volunteers living in areas with different levels of air pollution. We determined that particles with the size of 800 microns and higher of relatively favorable region, particles with the size of less than 50 microns dominated in the air of the unfavorable area, among which there were the most hazardous to health amounts of microparticles - from 200 to 300 nm. Microparticles of unfavorable area causes the development of oxidative modifications of proteins and DNA contributing to the change of leukocyte potential energy. The increase in total antioxidant activity and response of thiol-disulfide system (the increase in thioredoxin, glutathione with a stable reductase level] maintains a balance of oxidation and antioxidant processes contributing to protection of the cellular and subcellular structures against considerable oxidative damage

CLINICAL AND IMMUNOLOGICAL COMPARISONS IN Th- DEPENDENT IMMUNE RESPONSE MECHANISMS AMONG PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE

Chronic obstructive pulmonary disease (COPD) is a global public health problem. Studies in immunological features and their correlations with clinical course of COPD are of importance. The aim of this study was to elucidate clinical and immunological features in COPD of different severity grade, concerning Th1- and Тh17-dependent types of immune response.The study included 132 COPD patients and 32 healthy individuals. According to clinical and functional patterns, the patients with COPD were divided into 3 groups, i.e., 36 cases (28%) of mild severity; 62 individuals (48%), of moderate severity, and 30 patients (23%) of severe clinical grade. We have performed both clinical and immunological evaluation of the patients. The Th1- and Th17-specific lymphocyte subpopulations were assessed according to the serum levels of cytokines, i.e. tumor necrosis factor (TNFα), IL-4, IL-10, IL-17A, IL-21, IFNγ, as well as transforming growth factor β1 (TGF-β1). We have also determined expression of IL-6R receptor (CD126+) on mature T lymphocytes (CD3+) and T helper cells (CD4+) from peripheral blood. We have obtained the following results: the patients with mild-grade COPD exhibited three different T cell phenotypes were determined, with a prevalence of Th1-dependent immune response. The IL-6R were mostly expressed on CD3+CD126+ cells for the Th1/Th17 phenotype, and CD4+CD126+ cells in cases of Th17-dependent type immune response. In patients with COPD of moderate severity, the Th1, Th17, or Th1/Th17 types of immune response was revealed at similar rates. The level of IL-6R expression on mature T lymphocytes and T-helper cells increased to the greatest extent in cases of Th17-dependent immune response. In severe COPD patients, we have found a dominance of Th17 and Th1/Th17 type immune response. The levels of IL-6R expressionwere increased in Th17- and Th1/Th17-dependent types of immune response, the most significant increase was observed for CD4+ cells in Th17 phenotype. Clinical features of COPD proved to be associated with the phenotypes of immune response. These results allow of specifying the inflammatory phenotype, predicting the course of chronic disease, and selecting appropriate therapy

FEATURES OF CYTOKINE PROFILE IN PATIENTS WITH BRONCHIAL ASTHMA COMBINED WITH OBESITY

Combination of bronchial asthma (BA) and obesity is a difficult-to-control phenotype. Studies of inflammatory process with respect to severity of the disease are important for understanding the potential influence of obesity on the BA clinical course. The objective of this study was to determine cytokine profile in patients with mild BA combined with obesity. The study involved fifty-three patients with partially controlled mild BA. The patients were recruited as volunteers and signed an informed consent. The first observation group consisted of 27 asthma patients with normal body weight, the second observation group consisted of 26 patients with BA combined with obesity. A control group included 25 healthy volunteers. All the patients underwent clinical and laboratory examination in accordance with clinical standards for BA and obesity. The levels of TNFα, IL-2, IL-4, IL-6, IL-10 were evaluated in blood serum by means of flow cytometry. The ratios of proand anti-inflammatory cytokines (TNFα/IL-4, TNFα/IL-10, IL-6/IL-4, IL-6/IL-10) were calculated. Asthma patients with obesity (the 2nd group) had elevated levels of IL-2 over control group and group 1, by 38% and 44% respectively(p < 0.05). The concentration of proinflammatory cytokines TNFα and IL-6 was significanty increased in the both patient groups. Mean TNFα level was increased 2.5 times (p < 0.05), and IL-6 levels were increased by 30% (p < 0.05) in the 1st group as compared to the controls. TNFα and IL-6 concentrations showed a 3-fold increase over control values (p < 0.05) in the 2nd group. The level of antiinflammatory cytokine IL-4 was increased in patients with BA, independently of body mass. It should be noted that the concentration of this cytokine in obese patients was higher by 29% than in patients with normal body weight. IL-10 levels in patients from the 2nd group were reduced more than 2 times than in the 1st group. The patients of the 1st group showed a decrease in the IL-6/IL-10 index, in comparison with control parameters, thus indicative of an imbalance due to the elevation of the anti-inflammatory IL-10 cytokine. Among BA patients with obesity (group 2) the TNFα/IL-10 and IL-6/IL-10 indexes were higher than those of the control group (2.3- and 5.5-fold, respectively) and the group 1 (2.6- and 2.5-fold, respectively). Dynamics of these indexes confirms the systemic nature of inflammation and a predominance of non-atopic  inflammation in asthma patients with obesity. Thus, features of the cytokine profile in BA with obesity consist of a significant increase in pro-inflammatory IL-2, IL-6, TNFα cytokines, and a relative decrease in anti-inflammatory IL- 10 cytokine. The development of BA with obesity, even in mild-severity BA, is accompanied by development of a cytokine disbalance, which is typical for a mixed-type inflammation, with a prevalence of neutrophil inflammation

Features of immune response in different phenotypes of chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) is considered a heterogeneous disorder exhibiting different phenotypes. Chronic systemic inflammation is an important link in the COPD pathogenesis. The studies of immune response in the context of clinical and functional phenotypes seems relevant. Objective of our work was to study the features of immune response in clinical and functional phenotypes of COPD.Eighty-three COPD patients of different severity grade and 22 apparently healthy volunteers were examined. After determining the COPD phenotype by clinical and functional signs, the patients were divided in two groups, i.e., 38 subjects with bronchitis, and 45 patients with emphysematous phenotype. Clinical, functional and laboratory research was carried out in standard mode. Static lung volumes and respiratory capacities were investigated, i.e., functional residual capacity, residual lung volume, total lung capacity, bronchial resistance on inspiration and expiration to assess phenotype of the disease. Subpopulations of Th1 and Th17 lymphocytes were determined by the level of blood serum cytokines, tumor necrosis factor (TNFá), interleukins (IL) IL-4, IL-10, IL-17A, IFNã).Different features of immune response were revealed in bronchitic and emphysematous phenotypes of the COPD patients. Activation of inflammatory process with differentiation of naive T lymphocytes along the Th1-dependent pathway was found in 68% of cases with bronchitis and 16% of patients with emphysematous phenotypes. As compared with control group, the patients showed a statistically significant increase in the level of TNFá, IFNã, along with decrease in IL-4. Development of immune response by the Th17 type was found in 32% of cases with bronchitis, and 84% of cases with emphysematous phenotypes. Its emergence was associated with increased IL-17A and IL-10 levels, and a decrease in IFNã/IL-17A compared to the control. Differentiation of T helper cells towards Th1 pathway of immune response has been shown to predominate in bronchitic phenotype and at early stages of the disease. The Th17 type of immune response prevailed with increasing severity of the disorder. In emphysematous phenotype of COPD, the Th17-pathway of immune response develops at early stages of the disease. Some relationships are revealed between the systemic inflammation indexes and functional parameters of external respiration. An inverse relationship between TNFá and the OOL/OEL ratio in Th1 type of immune response has been shown. A direct correlation was found between the level of IL-17A and the parameters of external respiration function (FEV1, FEV1/FVC), as well as between IFNã/IL-17A and functional residual capacity in Th17 type of immune response.The type of immune response is associated with severity of the disease, as well with clinical and functional phenotype of COPD. Progression of the disease, broncho-obstructive disorders and hyperinflation are associated with increased levels of cytokines that provide cell polarization along the Th17 pathway. Determination of COPD phenotype and the type of immune response already at an early stage of the disease will enable prediction of its course and justify the choice of phenotype-oriented therapy

Патогенетические особенности мембранных нарушений иммунокомпетентных клеток при сочетанном течении бронхиальной астмы и хронической обструктивной болезни легких

The objective of this study was to investigate a role of fatty acid content in leukocyte membranes, oxylipin level and membrane potential of leukocyte membranes in the pathogenesis of immune cell membrane abnormalities in patients with comorbidity of asthma and chronic obstructive pulmonary disease (COPD).Methods. The study involved 39 patients with COPD, 41 patients with asthma, and 18 patients with comorbidity of COPD and asthma, and 28 healthy volunteers as controls. Fatty acid content of leukocyte membranes was investigated using gas-liquid chromatography. Thromboxane B2 and leukotriene B4 levels were measured using enzyme immunoassay. Mitochondrial membrane potential in leukocytes was measured ex tempore by cytofluorimetry. Statistically significant difference between mean values was determined by Student's t-test.Results. The comorbidity of COPD and asthma was characterized by accumulation of saturated fatty acids (12 : 0; 16 : 0; 18 : 0; 20 : 0) in leukocyte membrane and reduction in n-6 and n-3 polyunsaturated fatty acids (PUFAs) (18 : 2n-6; 20 : 3n-6; 20 : 4n-6; 20 : 5n-3; 22 : 4n-6; 22 : 6n-3). Changes in leukocyte lipidome contribute to development of structural and functional abnormalities of the cells and to the synthesis of oxylipins. This is confirmed by increased number of cells with reduced mitochondrial membrane potential and increased level of proinflammatory mediators, such as thromboxane B2 and leukotriene B4.Conclusion. Structural abnormalities of immune cell membrane should be considered as the primary pathological pathway to general dysfunction of the immune system and the basic mechanism of development of respiratory comorbidityКоморбидное течение хронической обструктивной болезни легких (ХОБЛ) и бронхиальной астмы (БА) является одной из важных проблем в пульмонологии, при этом многие патогенетические, диагностические и терапевтические аспекты указанной проблемы остаются нерешенными.Целью исследования явилось установление патогенетических особенностей мембранных нарушений иммунокомпетентных клеток при сочетанном течении ХОБЛ и БА на основании изучения состава жирных кислот мембран лейкоцитов, уровня оксилипинов, мембранного потенциала митохондрий лейкоцитов.Материалы и методы. В исследовании принимали участие больные ХОБЛ I и II спирометрического класса стабильного течения (n = 39), БА легкой степени тяжести контролируемого и частично контролируемого течения (n = 41), а также лица с коморбидным течением ХОБЛ I и II спирометрического класса и БА легкой степени тяжести контролируемого и частично контролируемого течения (n = 18); контрольную группу составили здоровые добровольцы (n = 28). Исследование состава жирных кислот мембран лейкоцитов проводилось методом газожидкостной хроматографии. Содержание тромбоксана В2 (ТХВ2) и лейкотриена В4 (ЛТВ4) определялось иммуноферментным методом. Уровень мембранного потенциала митохондрий лейкоцитов исследовался ex tempore методом цитофлуориметрии. Статистическая значимость различий средних величин определялась по t-критерию Стьюдента.Результаты. У пациентов с сочетанным течением ХОБЛ и БА в мембране лейкоцитов происходит накопление насыщенных жирных кислот (12 : 0, 16 : 0, 18 : 0, 20 : 0) на фоне дефицита n-6 и n-3 полиненасыщенных жирных кислот (18 : 2n-6, 20 : 3n-6, 20 : 4n-6, 20 : 5n-3, 22 : 4n-6, 22 : 6n-3). Изменения в липидоме лейкоцитов способствуют нарушению структурных и функциональных характеристик клетки, синтезу оксилипинов, что подтверждается увеличением числа клеток со сниженным мембранным потенциалом митохондрий и уровня провоспалительных медиаторов – ТХВ2 и ЛТВ4.Заключение. Нарушение архитектуры цитомембраны иммунокомпетентных клеток является первичным патогенетическим звеном в дисфункции работы всей иммунной системы, фундаментальной основой механизма развития коморбидной патологии органов дыхания

EFFICIENCY OF IMMUNOCORRECTION WITH MEDICAL OZONE IN PROTRACTED PURULENT INFLAMMATORY DISEASES AFFECTING SOFT TISSUES IN THE PATIENTS OF SENIOR AGE GROUP

Abstract. We performed a clinical and immunological study of patients from senior age group suffering with protracted purulent inflammation of soft tissues in maxillo-facial area. The patients underwent either common medication, or combined treatment including ozone therapy. Application of medicinal ozone, along with basic treatment schedule, resulted into more marked and rapid normalization of non-specific resistance markers, Т-cell immunity. This approach allows of reducing the terms of inpatient care and variety of possible complications. (Med. Immunol., 2008, vol. 10, N 2-3, pp 277-282)

CHANGES IN THE SURFACE IL-6 RECEPTOR EXPRESSION OF DISTINCT IMMUNE CELLS IN PROGRESSION OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE

Introduction: Mechanisms of immune response in COPD are still poorly understood, despite global challenge which this disorder presents to public medicine. The objective of this study was to evaluate IL-6R expression on the surface of immune cells in the course of COPD progression. Materials and methods. The study included 112 patients with stable mild, moderate and severe COPD (spirometry classes 1,2, and 3). A control group included 32 healthy subjects. The COPD patients were in remission phase. Expression of IL-6R on the surface of T lymphocytes, T helper cells, monocytes and granulocytes was determined by flow cytometry. Results and Discussion. Progression of the disease was accompanied by increase in the relative amounts of CD126+ immunocompetent cells. The number of CD4+CD126+ cells was maximally increased at all stages of COPD. Moreover, the number of CD126+ granulocytes proved to be significantly increased with increasing severity of COPD. Hence, it may be assumed that T-helper cells and granulocytes are primary target cells for IL-6 anti-inflammatory action, which is mediated by classic signaling