The Role of Foxp3-Expressing Regulatory T Cells and T Helpers in Immunopathogenesis of Multidrug Resistant Pulmonary Tuberculosis

Subpopulation structure of regulatory T cells and T helpers of peripheral blood in patients with newly diagnosed pulmonary tuberculosis depending on the clinical form of disease and sensitivity of Mycobacterium tuberculosis to antituberculosis drugs has been analyzed in this work. It has been shown that the leading part in immune suppression at infiltrative, dissemination, and fibrosis-cavity pulmonary tuberculosis is played by natural regulatory CD4+CD25+Foxp3+-T lymphocytes. Thus we estimate increase of their number in blood by drug-resistance and drug-susceptible patients. It has been demonstrated that in patients with fibrocavernous and infiltrative form of the disease and drug-resistant pulmonary tuberculosis the number of CD4+CD25−Foxp3+-regulatory T cells was increasing. In patients with infiltrative pulmonary tuberculosis, including multidrug-resistant M. tuberculosis, an increased number of CD3+CD4+CD25− T helpers is determined by the pathogenic features of the development of the tuberculosis infection and is connected with the activation of Th1-dependent immune response. Reduction in the number of T-helpers in the blood of patients with dissemination and fibrosis-cavity pulmonary tuberculosis mediates inefficient implementation of cell-mediated protective immunity

Single-shot optical conductivity measurement of dense aluminum plasmas

The optical conductivity of a dense femtosecond laser-heated aluminum plasma heated to 0.1-1.5 eV was measured using frequency-domain interferometry with chirped pulses, permitting simultaneous observation of optical probe reflectivity and probe pulse phase shift. Coupled with published models of bound-electron contributions to the conductivity, these two independent experimental data yielded a direct measurement of both real and imaginary components of the plasma conductivity.DOE National Nuclear Security Administration DE-FC52-03NA00156Physic

X-Ray Generation from Metal Targets Coated with Wavelength-Scale Spheres

X-ray yield measurements from targets coated with wavelength-scale spheres are compared with measurements from polished targets. Evidence for a hotter resonant electron temperature due to field enhancements from Mie resonances in the spheres is investigated

Control of Strong-Laser-Field Coupling to Electrons in Solid Targets with Wavelength-Scale Spheres

Irradiation of a planar solid by an intense laser pulse leads to fast electron acceleration and hard x-ray production. We have investigated whether this high field production of fast electrons can be controlled by introducing dielectric spheres of well-defined size on the target surface. We find that the presence of spheres with a diameter slightly larger than half the laser wavelength leads to Mie enhancements of the laser field which, accompanied by multipass stochastic heating of the electrons, leads to significantly enhanced hard x-ray yield and temperature

FUNCTIONAL POLYMORPHISM OF THE PRO-INFLAMMATORY CYTOKINE GENES IN PULMONARY TUBERCULOSIS

In the present time, incidence of pulmonary tuberculosis (TB) becomes broader, due to spreading resistance of Mycobacterium tuberculosis (MBT) to anti-tuberculosis drugs and infection with highly virulent strains of M. tuberculosis. The MBT antigens can cause dysfunction of the receptors and modulate the cytokine secreting function of immunocompetent cells. Polymorphic genes of pro-inflammatory cytokines involved in the mechanisms of defense responses of innate immunity, determine the degree of resistance to individual mycobacterial infection, as well as severity and duration of the disease in cases of clinical manifestations. The aim of the study was to investigate the connections between allelic polymorphisms of IL2, IFNG and TNFA genes and changes in secretion of the corresponding pro-inflammatory cytokines IL-2, IFNγ, and TNFα in vitro in patients with the newly diagnosed pulmonary tuberculosis (TB), depending on the clinical form of the disease.A total of 334 patients (220 men and 114 women) aged 23 to 50 years with newly diagnosed infiltrative and disseminated TB were enrolled into the study. The control group consisted of 183 healthy donors (130 men and 53 women) of corresponding age. The material of the research included DNA extracted from the whole blood and supernatants of culture suspensions of mononuclear leukocytes isolated from venous blood in healthy volunteers and patients with TB. The evaluation of cytokines secretion was performed by measuring their concentration in the blood mononuclear cell culture supernatants. using enzyme-linked immunosorbent assay (ELISA). To study polymorphic regions of cytokine genes, a polymerase chain reaction (PCR) was applied. Analysis of the obtained data was carried out by means of the program Statistica for Windows Version 6.0 (StatSoft Inc., USA).It was found that the imbalance of secretion of pro-inflammatory cytokines in TB patients was associated with the polymorphic variants of genes of these cytokines. It was found that the hypo-secretion of IL-2 is determined by the carriage of the G allele and genotype GG (T-330G) of the IL2 gene in both the control group and in patients with TB, regardless of the clinical form. In patients with DTB carriers of the homozygous genotype TT (T-330G) of the IL2gene, increased protein secretion was established. The maximum secretion of TNFб was recorded in patients with the AA genotype (G-308A) of the TNFA gene in the control group and in ITB patients; the minimum concentration of TNFα was associated with the carrier of the homozygous GG genotype (G-308A) of the TNFA gene in all the examined groups. In patients with ITB and DTB, an increase in IFNγ secretion by mononuclear blood leukocytes is not associated with the carrier of polymorphism +874A/T of the IFNG gene.Reduced secretion of IL-2 and TNFα in TB patients is associated with polymorphisms of their genes – (T-330G) of IL2 gene and (G-308A) of TNFA gene, respectively. The polymorphism (+874A/T) of the IFNG gene does not have a modulatory effect on the secretion of IFNγ in patients with TB, regardless of clinical form of the disease

Факторы иммуносупрессии при различных патологиях

Until recent time it has seemed obvious that suppressive function in the immune system is provided by one subpopulation of Tlymphocytes-suppressors. At present it is usually considered that regulatory cells (T-reg) are key cells-suppressors of the immune response. There exist two main mechanisms of T-reg immunosuppression realization: direct (when there is direct contact between cells) and distant (cytokine-dependent). For suppression of the immune response Т-reg cells produce cytokines with suppression activity: TGF-β, IL-10, IFN-γ, IL-35. Meanwhile the increasing number of facts indicates that suppression of the immune response is a multi-component process. A considerable role in suppression of the immune response is assigned to the endocrine system. However, immunosuppression mechanisms under infection, neoplastic processes and the influence of xenobiotics on the organism are not completely clear.До недавнего времени казалось очевидным, что супрессирующая функция в иммунной системе обеспечивается одной субпопуляцией Т-лимфоцитов-супрессоров. В настоящее время принято считать, что ключевыми клетками-супрессорами иммунного ответа являются регуляторные Т-клетки (T-reg). Существуют два основных механизма реализации иммуносупрессии T-reg: прямой (при непосредственном контакте между клетками) и дистантный (цитокинзависимый). Для супрессии иммунного ответа Т-reg выделяют трансформирующий фактор роста β, интерлейкины-10, -35, интерферон-γ. При этом все большее количество фактов указывает на то, что супрессия иммунного ответа является многокомпонентным процессом. Значительная роль в супрессии иммунного ответа отводится эндокринной системе. До конца не ясными остаются механизмы иммуносупрессии при инфекции, неопластических процессах, воздействии на организм ксенобиотиков

Production of angiogenesis mediators and the structure of the vascular wall in the heart in ischemic cardiomyopathy

Background. In the pathogenesis of ischemic cardiomyopathy (ICMP), angiopoiesis remains unexplored.The aim. To describe the vasculature of the heart and the imbalance of angiogenesis mediators in the coronary circulation in association with the number of endothelial progenitor cells (EPC) and desquamated endothelial cells (DEC) in the blood of patients with coronary heart disease (CHD), suffering and not suffering from ICMP.Methods. Fifty-two patients with CHD (30  patients with ICMP, 22  patients without  ICMP), 15  healthy donors were examined. The content of EPC (CD14+CD34+VEGFR2+) in the blood from the cubital vein and DEC (CD45–CD146+) in the blood from the coronary sinus and the cubital vein was determined by flow cytometry. The concentrations of VEGF-A (vascular endothelial growth factor A), PDGF (platelet-derived growth factor), and SDF-1 (stromal cell-derived factor 1) in blood plasma were recorded using immunofluorescence assay; the angiopoietin-2, MMP-9 (matrix metallopeptidase 9) were recorded using enzyme immunoassay. In myocardial biopsies the specific area of vessels and the expression of αSMA (smooth muscle alpha-actin) were determined by morphometric and immunohistochemical methods.Results. In the peripheral blood of patients with CHD, regardless of the presence of ICMP, the DEC content exceeded the physiological level, and the VEGF-A, PDGF, angiopoietin-2, and MMP-9 corresponded to the norm. In CHD patients without cardiomyopathy, there was an excess of SDF-1 and EPC in the blood from the cubital vein, and in ICMP, their physiological significance was noted. In the coronary blood flow in patients with CHD without cardiomyopathy, an increase in the concentration of PDGF was found, which was not determined in patients with ICMP, who had an increased content of DEC, angiopoietin-2 and MMP-9. The specific area of the vessels in the patients of the two groups was comparable; the expression of αSMA in ICMP was 6.2 times lower than in patients with CHD without cardiomyopathy.Conclusion. The development of ICMP is accompanied by impaired maturation of vessels in the myocardium, associated with the absence of a compensatory reaction of activation of cellular and humoral factors of angiogenesis

Expression of CD80 and HLA-DR molecules on blood monocytes in patients with pulmonary tuberculosis

We examined expression pattern of CD80 and HLA-DR pro-inflammatory molecules on the monocytes in patients with pulmonary tuberculosis (TB), depending on the clinical form of the disease and susceptibility of the pathogen to anti-tuberculosis drugs. The study involved forty-five patients with newly diagnosed pulmonary TB (25 men and 20 women aged 18 to 55 years, average age — 44.0±12.4 years). The control group included 15 healthy donors with similar socio-demographic characteristics as in TB patients. Venous blood was used as biomaterial for assays. Studies of the monocyte immunophenotype were carried out by flow cytometry of whole blood cells using Cytoflex flow cytometer (Beckman Coulter, USA) with specific monoclonal antibodies (eBioscience, USA). We determined the content of cells expressing surface markers of monocytes, i.e., CD14, CD45, CD80, and HLA-DR. The results of this study were evaluated using SPSS Statistics 17.0 standard software package and Microsoft Excel. In the course of the study, we have suggested a working hypothesis that the monocytes in TB patients, still being in circulation, can express activation markers during their migration to inflammation focus, especially CD80 and HLA-DR molecules. Analysis of the total CD14+ monocyte number showed its decrease in all forms and variants of clinical course of pulmonary tuberculosis compared with the control group. Assessment of pro-inflammatory markers expressed on CD14 positive monocytes, i.e., HLA-DR activation marker and CD80 co-stimulatory molecule, showed that the number of monocytes with HLA-DR expression in all TB patients was higher than in healthy donors. HLA- DR expression on CD14+ monocytes in the group of patients with infiltrative TB proved to be 15% higher than in patients with disseminated TB. The expression of CD80 on CD14+ monocytes in TB patients showed no differences between the groups and varied within the normal range. Hence, an imbalance within monocyte population in patients with pulmonary tuberculosis, regardless of its clinical form and drug sensitivity of the pathogen is developed, due to decrease in total number of CD14+ cells, along with increased relative number of monocytes expressing HLA-DR activation marker (pro-inflammatory phenotype). Meanwhile, expression of the CD80 co-stimulatory molecule on monocytes was within normal values

ПОЛИМОРФИЗМ ГЕНОВ ИММУНОСУПРЕССОРНЫХ ЦИТОКИНОВ IL-10 И TGF-β ПРИ ТУБЕРКУЛЕЗНОЙ ИНФЕКЦИИ

The aim of the work was the study of connection of allelic polymorphism of IL10 and TGFВgenes with changes in the basal and BCG-induced production of immunosuppressive cytokines IL-10 and TGF-β by mononuclear leukocytes in vitro in patients with the first diagnosed pulmonary tuberculosis (TB), depending on the clinical form of the disease. The evaluation of the cytokines production was conducted by measuring its concentration in culture supernatants by ELISA. The allele-specific amplification of specific stretches of the genome was used for the study of polymorphic genes of cytokines. The DNA and supernatants of culture suspensions of blood mononuclear leucocytes in healthy volunteers and patients with TB were the material of the research. It was shown in the research conducted that the basal and BCG-induced over-production of IL-10 in vitro occurs in patients with TB, regardless of the genotype of the locus of C-592AIL10 gene. In addition, genotype AA of polymorphism of IL10gene in patients with infiltrative and disseminated TB is associated with the maximum production of IL-10 in vitroand genotype CC – with the minimum production of this cytokine in vitro. Analysis of the production of TGF-β in vitro in patients with TB showed its increase only in case of carriage of allele T (C-509T) of TGFB gene. In patients with disseminated TB and homosygotic genotype TT the increase in both basal and BCG-induced production of TGF-β was determined, and in patients with infiltrative TB – only after induction of cells by BCG-antigen.Thus, the over-production of cytokines with inhibiting activity in patients with TB is genetically determined and promotes the formation of suppressive mode of immune-regulation. The increase in the secretion of cytokines IL-10 and TGF-β in vitro in patients with TB are associated with carriage of allele A and genotype AA (C-592A) of IL10gene and allele T and genotype TT (C-509T) of TGFB gene.Целью работы явилось исследование связи аллельного полиморфизма генов IL10 и TGFВс изменениями базальной и BCG-индуцированной продукции соответствующих иммуносупрессорных цитокинов IL-10 и TGF-β мононуклеарными лейкоцитами in vitro у пациентов с впервые выявленным туберкулезом легких (ТБ) в зависимости от клинической формы заболевания. Оценку продукции цитокинов осуществляли путем измерения их концентрации в культуральных супернатантах методом твердофазного иммуноферментного анализа (ELISA).Для исследования полиморфных участков генов цитокинов использовали аллельспецифичную амплификацию специфических участков генома. Материалом исследования являлись ДНК и супернатанты культуральных суспензий мононуклеарных лейкоцитов, выделенных из периферической венозной крови у здоровых добровольцев и больных ТБ. В ходе проведенных исследований выявлено, что базальная и BCG-индуцированная гиперпродукция IL-10 in vitro определяется при ТБ вне зависимости от генотипа локуса С-592А гена IL10. Наряду с этим у больных инфильтративным и диссеминированным ТБ генотип АА полиморфизма гена IL10 ассоциирован с максимальной, а генотип CC – с минимальной продукцией IL-10 in vitro. Анализ продукции TGF-β in vitro у больных ТБ показал ее увеличение только в случае носительства аллеля Т(С-509Т) гена TGFB. При этом у больных диссеминированным ТБ с гомозиготным генотипом ТТ фиксировалось увеличение как базальной, так и BCG-индуцированной продукции TGF-β, а у больных инфильтративным ТБ – только при индукции клеток антигеном BCG.Таким образом, гиперпродукция цитокинов с ингибирующей активностью у больных ТБ является генетически детерминированной и способствует формированию супрессорного режима иммунорегуляции. При этом выраженное увеличение секреции цитокинов IL-10 и TGF-β in vitro у больных ТБ ассоциировано с носительством аллеля А и генотипа АА (С-592A) гена IL10 и аллеля Т и генотипа ТТ (С-509Т) гена TGFB

Роль γδТ- и NK-клеток в иммунном ответе

The article presents up-to-date literature data on the role of γδТ-cells and different NK-cells subpopulations in the immune response. Their origin, immunophenotypes, range of secreted cytokines as well as effector functions and factors having stimulating and inhibitory effect on γδТ- and NK-cells are described. Mechanisms of antibacterial action of this type of cells are analyzed. Представлены современные литературные данные о роли γδТ-клеток и различных субпопуляций NK-клеток в иммунном ответе. Описываются их происхождение, иммунофенотипы, спектр секретируемых цитокинов, эффекторные функции и факторы, оказывающие стимулирующее и ингибиторное влияние на γδТ- и NK-клетки. Анализируются механизмы антимикробного действия клеток данных типов