Молекулярные аспекты функционирования T-хелперов 17-го типа

The article presents up-to-date data on the functioning of the T-helper type 17 (Th17). History of the Th17 discovery, transcription factors, mechanisms, mediating stimulation and inhibition of the Th17 differentiation, are described. Cytokines, which are produced by this lymphoid subset, and signaling pathways, through which its effects are realized, are listed. A brief description is given of the main physiological processes and diseases, in which participation of the Th17 was described.Представлены современные научные данные о функционировании Т-хелперов 17-го типа (Th17). Описывается история открытия Th17, транскрипционные факторы, а также механизмы, опосредующие стимуляцию и ингибирование дифференцировки Th17. Перечислены цитокины, продуцируемые данной лимфоцитарной субпопуляцией, и сигнальные пути, за счет которых реализуются их эффекты. Дается краткое описание физиологических процессов и заболеваний, в развитии которых доказано участие Th17

Serum cytokines levels in patients with myocardial infarction with non-obstructive and obstructive coronary arteries

Aim. To compare the concentrations of proinflammatory and anti-inflammatory cytokines in patients with myocardial infarction with non-obstructive (MINOCA) and obstructive coronary arteries (MIOCA) in the early postinfarction period and after 1-year follow-up.Material and methods. The study included 40 patients with myocardial infarction (experimental group, 19 patients; control group, 21 patients). Three (15,7%) patients with diagnosed acute myocarditis were excluded from the final analysis. Blood samples were taken upon admission, on the 2nd, 4th and 7th days from hospitalization, and also after 1-year follow-up. Twenty-three parameters were analyzed using multiplex analysis and the Multiplex Instrument FLEXMAP 3D system (Luminex Corporation), as well as the MILLIPLEX map Human Cytokine/ Chemokine Panel II.Results. According to multiplex analysis of blood serum of the studied groups, a comparable increase in proinflammatory cytokines CCL-15, CCL-26, CCL-27 in the early postinfarction period and after 1-year follow-up, as well as antiinflammatory and regenerative cytokines CXCL-12, TPO in the early postinfarction period and after 1-year follow-up. In patients with MINOCA, higher concentrations of the following proinflammatory cytokines were determined: IL-16 upon admission (p=0,03), IL-20 on days 2 and 4 of the early postinfarction period (p=0,005 and p = 0.03), as well as CCL-15 on days 4 and 7 (p=0,05 and p=0,02). After 1-year follow-up, among the proinflammatory cytokines, a greater increase in CCL-21 (p=0,02) was noted in the patients of experimental group. Also, in patients with MINOCA, a greater increase in TPO was determined upon admission and on the 2nd day (p=0,02 and p=0,02), SCF — on the 7th day and after 1-year follow-up (p=0,04 and p=0,04), and LIF on the 4th day of early postinfarction period (p=0,007). In contrast, MIOCA patients showed a greater increase in CXCL-12 levels upon admission (p=0,04). At the same time, patients with MINOCA showed a higher level of C-reactive protein on the 1st day, as well as a higher relative monocyte count after 1-year follow-up.Conclusion. Despite a comparable increase in the cytokines CCL-8, CCL-13, CCL26, CCL-27 in patients of both groups, in patients with MINOCA there was a greater increase in proinflammatory cytokines IL-16, IL-20, CCL-15, CCL-21, and also CXCL-12, LIF, TPO, SCF, which have anti-inflammatory and regenerative activity. After 1 year follow-up, MINOCA patients showed a significant increase in CCL-21 and SCF, with a comparable increase in other proinflammatory cytokines in patients of both groups. A greater increase in proinflammatory cytokines in patients with MINOCA may indicate a more aggressive atherosclerosis course and lead to plaque destabilization followed by ischemic event

Hypersegmentation of neutrophil nuclei in peripheral blood of patients with localized and advanced cancer of the larynx and laryngopharynx

Neutrophilic granulocytes have a wide spectrum of functional activity. In recent years, the functional significance of neutrophils in the development and course of malignant neoplasms has been discussed. It has been shown that neutrophilic granulocytes can play pro- or antitumor activity. The aim of the study was to assess the structural and functional features of neutrophils in patients with varying degrees of prevalence of cancer of the larynx and laryngopharynx. Forty-one patients (aged 35-67) with newly diagnosed cancer of the larynx and laryngopharynx were examined and divided into subgroups according to the TNM classification: the first subgroup (14 patients) with a localized tumor process consisted; and the second subgroup (27 patients) with a widespread tumor process. The relative and absolute number of neutrophils was assessed, and the neutrophil-lymphocyte ratio (NLR) was determined. The content of neutrophils with varying degrees of nuclear segmentation in the blood was calculated, the activity of myeloperoxidase, cationic proteins, alkaline phosphatase, and the degree of neutrophil activation in the NBT test was determined cytochemically. Concentration of interleukin-8 was determined using ELISA. In patients with cancer of the larynx and laryngopharynx the number of neutrophils (p = 0.045) and NLR (p = 0.033), as well as serum concentration of interleukin 8 (p = 0.011), increased compared to healthy individuals. The proportion of cells with hypersegmented nuclei in the neutrophil population (p < 0.001) and cytotoxic potential increased with the spread of tumor process. A direct correlation (r = 0.42, p = 0.03) was found between the T index, which reflects the volume of the tumor, and the content of hypersegmented neutrophils. It can be argued that such a simple and accessible laboratory parameter as the degree of segmentation of the nuclei of neutrophilic granulocytes can be used as one of the criteria to assess and predict the course of the tumor process

Eotaxin and cardio-ankle vascular index in patients with high and very high cardiovascular risk

Eotaxin is a chemokine, which is a chemoattractant mainly to eosinophils, as well as basophils and Th2 lymphocytes. According to studies, overexpression of eotaxin is found in endothelial and smooth muscle cells of blood vessels in the area of atherosclerotic plaque. In clinical medicine, cardio-ankle vascular index (CAVI) is widely used as an indicator of arteriosclerosis and a predictor of cardiovascular events. Few studies have shown the relationship of eotaxin with coronary atherosclerosis; in other studies, the relationship of eotaxin with atherosclerosis, myocardial infarction and pulse wave velocity was not revealed. The aim of the present study was to assess blood level of eotaxin and cardio-ankle vascular index and their association with major cardiovascular risk factors in patients with high and very high cardiovascular risk. We examined 65 patients with high and very high cardiovascular risk, due to documented coronary artery disease, type 2 diabetes mellitus, or combination of cardiovascular risk factors and who were undergoing generally accepted cardioactive, hypoglycemic therapy and lipid-lowering therapy. All patients were examined for the elastic properties of the vascular wall by volumetric sphygmography with assessment of CAVI. In the blood, the concentrations of eotaxin, high-sensitivity C-reactive protein, glycosylated hemoglobin and lipid spectrum indicators were determined. All examined were divided into two groups: with a normal value of CAVI (less than 8) and elevated. Patients with elevated CAVI had higher concentrations of eotaxin (p = 0.013), total cholesterol (p = 0.009), low-density lipoprotein cholesterol (p = 0.016), were older (p < 0.0001) and less likely to take statins (p = 0.002). In all those examined, correlations were found between serum eotaxin concentration and CAVI (rs = 0.34; p = 0.005), as well as age (rs = 0.32; p = 0.006). The age of the patients correlated with CAVI (rs = 0.35; p = 0.007). Thus, in our study, we for the first time showed the relationship between higher concentrations of eotaxin and an increased cardio-ankle vascular index in patients with high and very high cardiovascular risk. Cardio-ankle vascular index was associated with age, lipid metabolism and lipid-lowering therapy. The obtained results allow us to consider eotaxin as a factor associated with atherogenesis and arterial stiffness

Effects of intranasal Imunofan administration upon phagocytic activity in treatment of exudative otitis media in children

Exudative otitis  media  in  childhood is most  often  associated with  chronic inflammation in the  nasopharyngeal area,  with  immediate participation of phagocytic cells. Our  paper  presents  the  data  on evaluation of clinical  and immunological efficacy of intranasal Imunofan use included into  complex therapy of exudative  otitis  media.  Dynamic observation (before  treatment, 1 and  3 months after treatment) of these parameters included regular  evaluation of the  neutrophil and  monocyte amounts in peripheral blood  and  in smear imprints from nasal mucosa, determination of myeloperoxidase activity in circulating neutrophils, and the content of interleukin IL-8 and IL-18  in the nasal washouts. The clinical status was assessed using a scoring system, which subjectively reflected the state of the nasopharynx and auditory function. Fourty-three children aged from 3 to 7 years with exudative  otitis media associated with chronic adenoiditis were examined. Patients of the first group (22 children) were treated using only conventional approaches (basic therapy). The patients from  the  second  group  (21 children) received  Imunofan in addition to the  basic therapy. The  control group consisted of 16 relatively  healthy  children. Before  treatment of the  children with exudative  otitis  media, an increase  in the relative content of monocytes in their blood,  a decreased activity of myeloperoxidase and lower concentration of IL-8  and  IL-18  in the  nasal  wash was observed  in comparison with  healthy  controls. No differences in severity  of clinical  symptoms were revealed  between  the  groups  of patients. Baseline  therapy was not  accompanied by positive  dynamics in the  clinical  pattern of the  disease.  Relative  monocytosis and reduced activity of neutrophilic myeloperoxidase persisted  in peripheral blood;  the concentration of IL-8  and IL-18  in the  nasal washings  remained low. Following intranasal use of Imunofan, the  number of circulating monocytes was restored by the  third  month from  the  start  of treatment, there  was an  increased activity  of myeloperoxidase registered  in blood neutrophils, as well as higher IL-8  and IL-18  concentrations in the nasal washings. Normalization of the phagocytos-related parameters, according to this scoring,  was associated with clinical remission of the disease. The revealed relationships between clinical data and the results obtained in the course  of laboratory research  suggest a positive effect of Imunofan as an agent that may enhance effectiveness of conventional basic therapy  of otitis media in children

Association of decreased glomerular filtration rate with renal hemodynamic disorders and inflammatory biomarkers in patients with medically-controlled hypertension of high cardiovascular risk

Aim. To assess markers of chronic kidney disease (CKD) in patients with medically-controlled hypertension (HTN) (<140/90 mm Hg), as well as to analyze potential association of decreased glomerular filtration rate (GFR) <60 ml/min/1,73 m2 with clinical data and therapy; to establish significant determinants of GFR decrease in this category of patients.Material and methods. The study included 70 patients with HTN and office blood pressure (BP) <140/90 mm Hg aged 64 (57; 68) years (men, 48,6%), of whom 40 patients were examined within the Russian multicenter CHRONOGRAPH program. Office BP was 130 (120; 140)/80 (72; 82) mm Hg. GFR and albuminuria were assessed. Twenty-four-hour BP monitoring and Doppler ultrasound of renal blood flow with estimation of resistance indices (RI) were performed. The content of highsensitivity C-reactive protein (hsCRP), interleukins (IL) 1β, 6, 10 and lipid transport function parameters was determined.Results. CKD markers were detected in 31,4% of patients (in 27,1% — a decrease in GFR <60 ml/min/1,73 m2; in 12,9% — pathological albuminuria). Patients with CKD markers were older, had higher office systolic BP and nocturnal pulse pressure, higher blood hsCRP and RI levels throughout the renal flow, and lower high-density lipoprotein cholesterol levels. In the presence of CKD markers, calcium channel blockers, aldosterone receptor antagonists and statins were used more often. The results of correlation analysis were used to determine the determinants of GFR decline. In the general group, GFR values had inverse correlations with age (Rs=-0,58, p<0,0001), segmental intrarenal artery RI (Rs= -0,4232, p=0,0005), blood hsCRP (Rs=-0,3998, p=0,0007), IL-1β (Rs=-0,3139, p=0,0086), office BP and some 24-hour BP parameters. In the presence of CKD markers, a direct association of GFR and IL-10 was determined (Rs=0,4293, p=0,046). In the absence of such markers, GFR had an inverse correlation with IL-1β content (Rs=-0,3110, p=0,0333). A multiple linear regression model included following independent determinants of GFR: age, blood hsCRP and RI in the segmental intrarenal arteries.Conclusion. Among patients with medically-controlled HTN of high cardiovascular risk, a high prevalence of CKD markers was revealed (31,4%). Compared with patients with preserved renal function, in the presence of CKD markers, there were higher levels of office systolic BP, nocturnal pulse pressure, blood hsCRP, and intrarenal artery RI. Associations were established between GFR and the levels of hsCRP, IL-1β and IL-10, which confirms the pathological role of inflammatory biomarkers in developing renal dysfunction in high-risk HTN. Age, elevated blood hsCRP levels, and intrarenal artery RI are independent determinants of decreased GFR in patients with medically-controlled HTN of high and very high cardiovascular risk. The data obtained shows the need for early prescription of combination antihypertensive therapy with nephro- and vasoprotective effects in this category of patients, as well as with an ability to depress the chronic subclinical inflammation

IMMUNOREGULATORY IMBALANCE AND FUNCTIONAL STATE OF THE HEART IN THE PATIENTS WITH DIABETES MELLITUS TYPE 2

Diabetes mellitus type 2 is one of the most important non-infectious diseases in the modern world, being an important risk factor of cardiovascular disorders. Changes in left ventricular myocardial diastolic function are observed in diabetic patients independently from other comorbidities. Etiology of the heart failure during diabetes mellitus type 2 is multifactorial, exhibiting cellular,  molecular and metabolic aspects. However, its pathophysiological mechanisms are not completely understood. The aim of this study was to evaluate numbers of inflammatory T lymphocytes, i.e., T helper type 1 (Th1) and T helper type 17 (Th17) cells, and FoxP3+T regulatory lymphocytes, depending on the functional state of the heart assessed by two-dimensional echocardiography in patients with arterial hypertension and diabetes mellitus type 2. A total of twenty-five patients with a combination of arterial hypertension and diabetes mellitus type 2, and 14 patients with arterial hypertension without carbohydrate disturbances were recruited to a cross-ectional case-control study. All the patients underwent echocardiography with transthoracic access at the M-mode, B-mode and Doppler mode of imaging. We evaluated numbers of Th1 and Th17 lymphocytes by intracellular production of IL-17 and IFNγ by CD4+ lymphocytes, respectively. The numbers of FoxP3+T regulatory lymphocytes were estimated by expression of CD25 and FoxP3 transcription factor. A flow cytometry approach was used in both cases. We revealed some correlations between the numbers of Th17 lymphocytes, FoxP3+T regulatory lymphocytes and functional parameters of myocardium in patients with diabetes mellitus type 2, which were absent in patientswithout carbohydrate impairments. The numbers of FoxP3+T egulatory lymphocytes, Treg/Th17 lymphocyte ratio, and mean fluorescence intensity of IL-17 for Th17 cells was lower in patients with diabetes mellitus and diastolic dysfunction compared to the patients with diabetes free of diastolic dysfunction. Association of diastolic dysfunction with diabetes mellitus type 2 was accompanied by increase of IFNγ+Th1 lymphocyte numbers and concentrations of IL-10, IFNγ and TNFα in serum as compared to the patients with diastolic dysfunction in the absence of carbohydrate metabolism disturbances. The diabetic patients with diastolic dysfunction were characterized by hyperinsulinemia, hyperglycemia, higher index of insulin resistance, increase of waist circumference and visceral adiposity index when compared to the patients with diastolic dysfunction without diabetes. Visceral obesity and decrease of insulin sensitivity may be regarded as pathogenetically significant factors for the development of immune regulatory imbalance and diastolic dysfunction in the patients with diabetes mellitus type 2

SUBCELLULAR LOCALIZATION OF FoxP3 TRANSCRIPTION FACTOR IN PATIENTS WITH ACUTE CORONARY SYNDROME: COMPARATIVE ANALYSIS AND PROSPECTIVE OBSERVATION

The key cellular and molecular factors being involved in the resolution of inflammation following acute myocardial infarction remain poorly understood. T-regulatory (Treg) lymphocytes are characterized by the extreme potential to regulate the strength and direction of immune responses during the myocardial injury. The functional activity of Treg-lymphocytes depends upon the transcription factor forkhead box protein P3 (FoxP3). It may be also expressed in conventional T-lymphocytes at the stage of their activation. Nuclear localization of FoxP3 is a prerequisite factor determining its ability to impact the suppressive functions of Treglymphocytes.The aim of the present study was comparative evaluation of FoxP3+T-lymphocytes frequency and counts, combined with estimation of FoxP3 subcellular localization, in patients with acute myocardial infarction and chronic coronary syndrome and examination of changes of these parameters in the short-term follow-up of patients with myocardial infarction. The study included 14 patients with chronic coronary syndrome (8 males; 6 females; 63.2±9.0 y.o.) and 5 patients with acute anterior ST-segment elevation myocardial infarction (4 males; 1 female; 61.4±11.2 y.o.) at days 1, 3 and 7 after the event. The frequency of FoxP3+ conventional and regulatory T-lymphocytes was evaluated in peripheral blood mononuclear cells together with estimation of the level of FoxP3 nuclear localization by imaging flow cytometry.Patients with infarction were characterized by the decreased counts of FoxP3+Treg-lymphocytes compared to patients with chronic coronary syndrome, and exhibited even further decrease in the counts of FoxP3+Tregcells at day 7 after infarction, while frequency of Treg and conventional T-lymphocytes did not differ significantly. The level of FoxP3 nuclear translocation was lower both in Treg and conventional T-lymphocytes in patients at day 1 post-infarction compared to patients with chronic coronary syndrome. Absolute counts of FoxP3+Tregs with nuclear FoxP3 localization remained significantly lower both at days 1 and 7 post-infarction compared to patients with chronic coronary syndrome.Thus, here we demonstrated that FoxP3 nuclear localization experiences decrease in the course of acute myocardial infarction and may serve as a more sensitive marker of changes in Treg-lymphocyte functioning than simple evaluation of their frequency

Магнитно-резонансная томографическая оценка абдоминальной жировой ткани и состояние брюшной аорты у пациентов с ишемической болезнью сердца: связь с нарушениями метаболизма

Aim. To evaluate potential associations between quantitative features of visceral and subcutaneous adipose tissue (AT) and anthropometric characteristics of obesity, metabolic disorders, and the state of the abdominal aorta in patients with chronic coronary artery disease (CAD).Materials and methods. The study included 55 patients (average age 61.2 ± 7.2 years) with chronic CAD. Magnetic resonance imaging (MRI) was performed on a 1.5 T MRI scanner using T2-weighted spin-echo modes. The area and volume of abdominal subcutaneous (SAT) and visceral adipose tissue (VAT) were calculated at the L4–L5 level; the total volumes of abdominal SAT and VAT were determined. Parameters of lipid and carbohydrate metabolism, as well as adipokine profile were studied in the blood serum.Results. In the course of a multiple linear regression analysis, we detected the independent determinants, which described 95% of the total VAT volume variability and were represented by waist circumference (WC) and serum levels of high-density lipoprotein (HDL) cholesterol and adiponectin. The model was characterized by the significance level p < 0.000001, the residuals of the model were normal. We calculated the coefficients in the model: 1.39 for WC, –0.26 for HDL cholesterol, and –0.19 for adiponectin. We detected a positive correlation between the abdominal aorta (AA) diameter and SAT area at the L4–L5 level (rs = 0.48; p = 0.0014), which does not depend on gender, and reverse correlations between the aorta diameter and glycated hemoglobin (HbA1c) level (rs = –0.40; p = 0.0359) and postprandial glycemia (rs = –0.40; p = 0.0273). The patients with a dilated aorta (group 2), when compared with the patients with a normal aorta diameter (group 1), did not differ in the AT accumulation, but demonstrated decreased levels of HbA1c and postprandial glycemia, which resulted in a smaller number of patients with type 2 diabetes mellitus.Conclusion. We identified independent determinants of an increase in the total volume of abdominal visceral AT, such as an increase in WC and a decrease in serum adiponectin and HDL cholesterol levels. Results of the study indicate the presence of a link between the AA remodeling, accumulation of subcutaneous AT, and impaired glucose metabolism.Цель. Изучение потенциальных взаимосвязей количественных характеристик висцеральной и подкожной жировых тканей (ЖТ) с антропометрическими показателями ожирения, нарушениями метаболизма и состоянием брюшного отдела аорты у пациентов с хронической ишемической болезнью сердца (ИБС).Материалы и методы. В исследовании приняли участие 55 пациентов (средний возраст 61,2 ± 7,2 лет) с хронической ИБС. Магнитно-резонансную томографию выполняли на 1,5 Т магнитно-резонансном томографе в Т2-взвешенных спин-эхо режимах. Расчет площади и объема абдоминальных подкожной (ПЖТ) и висцеральной ЖТ (ВЖТ) проводился на уровне L4–L5, а также измерялся общий объем абдоминальных ПЖТ и ВЖТ. В сыворотке крови исследовали показатели липидного и углеводного метаболизма и состояние адипокинового профиля.Результаты. В ходе множественного линейного регрессионного анализа установлены независимые детерминанты вариабельности общего объема абдоминальной ВЖТ, которые описывают 95% вариабельности этого показателя: окружность талии (ОТ), содержание в крови холестерола липопротеинов высокой плотности (ХС-ЛВП) и адипонектина. Уровень значимости модели составил p < 0,000001, остатки модели нормальны. Оценки коэффициентов в модели для: ОТ – 1,39; ХС-ЛВП – 0,26 и адипонектина – 0,19. Документированы корреляционные взаимосвязи диаметра абдоминальной аорты (АА): прямая – с площадью ПЖТ L4–L5 (rs = 0,48; p = 0,0014), что не зависит от пола, и обратные – с уровнем гликированного гемоглобина (HbA1c) (rs = –0,40; p = 0,0359) и постпрандиальной гликемией (rs = –0,40; p = 0,0273). В сравнении с пациентами, имеющими нормальные значения диаметра АА (группа 1), у пациентов с расширенной АА (группа 2) не было значимых различий накопления ЖТ, но имел место более низкий уровень HbA1c и поспрандиальной гликемии, что определялось меньшим числом пациентов с сахарным диабетом типа 2.Заключение. Установлены независимые детерминанты возрастания общего объема абдоминальной висцеральной ЖТ, которыми являются: увеличение окружности талии и сниженные значения содержания в крови адипонектина и ХС-ЛВП. Результаты исследования свидетельствуют о наличии взаимосвязи между процессами ремоделирования АА, накоплением подкожной абдоминальной ЖТ и нарушениями метаболизма глюкозы