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Produkce cytokinů v průběhu mastitid skotu
Tylčová, Natálie
The aim of this work was to detect the pro-inflammatory cytokine TNF-α and the anti-inflammatory IL-10. The experiment included 20 dairy cows at different stages of mastitis, which came from the Nová Ves agricultural cooperative. 80 samples were totaly taken, from all quarters, from each dairy cow, which were divided into three groups. The first group represented healthy dairy cows, without clinical signs of mastitis, with a somatic cell count of up to 100 000, from 5 dairy cows, that is 20 milk samples. The second group consisted of dairy cows with clinical signs of mastitis and somatic cell counts up to 400 000, from 10 cows, that is 40 milk samples. The third group consisted of dairy cows with somatic cell counts of more than 1 000 000, from 5 cows, that is 20 milk samples. These samples were processed in the laboratory of the Department of Animal Morphology, Physiology and Genetics at Mendel University in Brno. The concentration of TNF-α and IL-10 was detected by ELISA. The average concentration of TNF-α in the first group was 138.70 pg/ml, in the second group 133.60 pg/ml, in the third group 95.05 pg/ml. The average concentration of IL-10 in the first group was 9.52 pg/ml, in the second group 11.03 pg/ml, in the third group 8.66 pg/ml.
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Cytokines in the effector function of regulatory T cells
Zadražil, Zdeněk ; Holáň, Vladimír (advisor) ; Stříž, Ilja (referee)
Regulatory T cells (Treg) are an important control mechanism within the Immune system (IS). Tregs prevent overactivation of effector T cells or autoreactive cells from invading organism-derived tissues. Treg are characterised by expression of surface molecules, CD4, CD25 and by an intracellular transcription factor forkhead box protein 3 (FoxP3). There are two basic populations of Treg, naturally occuring Treg (nTreg) developing in the thymus and induced Treg (iTreg) rising from CD4+ T cells in periphery, which are also precursors for T helper cells. In spite of an outgoing intensive research, there is still no clear clue which mechanisms are used by Treg to inhibit other effector cells. First in vitro experiments showed, that those mechanisms are of a contact dependent manner and do not use secreted molecules. But in vivo experiments showed the exact opposite. Those studies showed that secretory molecules, such as interleukin (IL)-10, IL-35 or transforming growth factor beta (TGF-β), are important in the effectory phase of Treg. Since the first experiments other distinct mechanisms of supression by Treg cells have been discovered. Those mechanisms seem to be important only in particular situations, particular cell assays or with using of specific experimental models. The reasons for this...
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The role of TNF-alpha and IL-10 in cardioprotective effect of chronic hypoxia
Chytilová, Anna ; Bezouška, Karel (advisor) ; Hloušková, Patricie (referee)
The aim of the present study was to find out whether adaption to chronic hypoxia affects the expresion of TNF-α and IL-10 in rat myocardium. TNF-α is a proinflammatory cytokine, which amplifies inflammatory reaction, while IL-10 has opposite antiinflammatory effect. We also measured concentration of nitrotyrosine as a marker of nitrosative stress. We used male Wistar rats divided into four groups: 1) normoxic controls; 2) exposed to continous normobaric hypoxia (10% O2) for three days or 3) for three weeks and 4) exposed to intermittent normobaric hypoxia (10% O2) for three weeks with one hour daily reoxygenation. Cytosolic and membrane proteins (cytosolic and particulate fractions) were obtained from the left ventricle, right ventricle and interventricular septum. Concentrations of TNF-α and IL-10 in both fractions were measured by ELISA. Continous hypoxia increased TNF-α production in particulate fractions from all ventricular parts and decreased the ratio of IL-10/TNF-α in particulate and cytosolic fractions. Intermittent hypoxia redistributed TNF-α from cytosol into the particulate fraction and prevented the drop of IL-10/TNF-α ratio in the cytosolic fraction. The highest concentration of nitrotyrosine was found in the particulate fraction from the right ventricle after three days of hypoxia....
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Benigní helmint Hymenolepis diminuta pozitivně ovlivňuje chemicky vyvolanou kolitidu u potkaního modelu
LEVÁ, Jana
In this study is examined the protective effect of different life cycles stages of rat tapeworm Hymenolepis diminuta against dinitrobenzene sulphonic acid (DNBS) induced colitis in rat model. The clinical health of rats, gut microbiota and systemic inflammation was analysed. For determination of the level of inflammation and type 2 immune response of rat model was used relative gene expression of TNF, IL-1, IL-4, IL-13 and IL-10 using real-time PCR. Our results showed that colonization by immature life stages of H. diminuta before the induction of DNBS colitis reduced clinical symptoms and inflammation in rat model but did not protect rats against colitis. Type 2 immune response was detected by an increase in gene expression of IL-4, IL-10 and IL-13. The gut microbiota does not substantial role in H. diminuta-mediated protection.
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Mesenchymal stem cells and their effects on regulatory B cells
Smolová, Helena ; Boháčová, Pavla (advisor) ; Stříž, Ilja (referee)
Mesenchymal stem cells (MSC) are multipotent cells with the ability to regulate reactivity of cells of immune system. Regulatory B cells (Bregs) are also capable of modulating immune responses. Both these cell types are able of creating anti-inflammatory and tolerogenic environments and represent potential of cell-mediated therapy for autoimmune diseases and transplantation reactions. The effect of MSC on Bregs activation and function has been only studied in recent years, and mechanisms of their effects are not yet well characterized. However, studies have demonstrated a decrease in effector B lymphocytes and antibody production, and a support of activation of Bregs subpopulation and increased production of anti-inflammatory interleukin 10. Various molecules produced by MSC are involved in Bregs induction. Unfortunately, their effects have not yet been sufficiently described, and different models yields diverse results. In addition to the current studies in experimental models, the first clinical trials on Bregs have been initiated. The positive results suggesting the potential for future use of Bregs for the treatment of autoimmune diseases and transplantation reactions have been obtained in both cases. Key words: regulatory B cells, mesenchymal stem cells, immunomodulation, autoimmune diseases,...
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Determination of inflammatory markers of the eye based on the analysis of tears - potential and limits
Mandíková, Šárka ; Daňková, Pavlína (advisor) ; Sičáková, Silvia (referee)
In this study, we aimed to determine the levels of cytokines IL-1β, IL-4, IL-10, IFN-γ, MIF and VEGF in tears derived from healthy subjects. We tested cytokines as potential markers of inflammation for their potential use in clinical practice. Having reliable method for measuring cytokine levels in tears would enable an early diagnosis of eye diseases. In two phases, cytokines in tears of healthy individuals were analyzed using Bio-Plex Cytokine Assay (Bio-Rad). We assessed the suitability of methods for diagnostic purposes as well as the suitability of our selected cytokines. Statistically significant positive correlations of cytokines were confirmed: IL-10 with IFN-γ (r = 0,81), MIF with VEGF (r = 0,42 / r = 0,49), IL-1β with IL-10 (r = 0,52), IL-1β with IFN-γ (r = 0,55), IL-1β with VEGF (r = 0,38), IFN-γ with VEGF (r = 0,45) and IL-4 with VEGF (r = 0,48) in healthy subjects in tears. IL-4 (r = -0,37) and IFN-γ (r = -0,42) correlate negatively with age. In healthy individuals, there seem to be no differences with regard to gender, BMI, body fat, time of meal consumption prior to tear collection, eye strain when using a computer, dry eyes. Thus, studied cytokines are suitable for diagnostic purposes. Significant differences in concentrations of four (IL-1β, IL-10, IFN-γ a VEGF) of the five...
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Different capacity of in vitro generated monocyte-derived dendritic cells of newborns of healthy and allergic mothers to prime immune responses
Súkeníková, Lenka ; Hrdý, Jiří (advisor) ; Javorková, Eliška (referee)
(EN) Reduced microbial stimulation of an immature neonatal immune system can lead to a poor balance adjustment of immune responses, thus contributing to the development of allergic diseases, whose incidence continues to rise. One of the promising precautionary measures seems to be an early preventive administration of probiotic bacteria to pregnant or nursing mothers, or to newborns. Previous works have described a beneficial effect of Escherichia coli O83:K24:H31 (E. coli O83) in the prevention of allergic diseases. In order to contribute to the clarification of E. coli O83 effects on the neonatal immune system, its immune- modulating properties were tested in vitro on umbilical cord blood cells. The ability of E. coli O83 to support the maturation of in vitro-derived dendritic cells from cord blood precursors (moDCs) of the children of healthy (children with a relatively low risk of allergy) and allergic (children at a relatively high risk of developing allergies) mothers was tracked by flow cytometry, qPCR and ELISA. Probiotic bacteria-stimulated moDCs were subsequently cultured with autologous naive CD4+ T lymphocytes and immune response polarization was also characterised by flow cytometry, qPCR, and ELISA. It was evident from the results that E. coli O83 promoted moDCs maturation. The presence of...
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Autoimmune and lymphoproliferative diseases: associations and common mechanisms
Dobiášová, Alena ; Daňková, Pavlína (advisor) ; Hušáková, Markéta (referee)
Autoimmune and lymphoproliferative diseases share some etiologic mechanisms. The origin of the diseases is complicated process that involves an accumulation of hereditary and somatic mutations in a hematopoetic cell, which thanks to changed activity overcomes different growth and survival control checkpoints. Such mutations are for example those located in genes coding for transcription factors, apoptotic signaling molecules, costimulatory molecules and secreted exctracellular molecules. All these molecules influence the balance between survival and programmed cell death. Their dysregulated expression enables the cell to overcome defensive mechanisms of the immune system. Therefore, autoimmune and malignant cells are able to survive though, under usual circumstances, they would be selected. The main aim of this work is to shed the light on the influence of the dysregulated expression of the particular molecules on the origin of autoimmune and lymphoproliferative diseases. Key words: autoimmune ilnesess, lymphoproliferative diseases, etiology, AIRE, c-MYC, TP53, FOXP3, Fas, PTEN, Bim, CTLA-4, CD5, CD30, CD40/CD40L, BAFF, α-taxilin, IL- 10.
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Cytokines in the effector function of regulatory T cells
Zadražil, Zdeněk ; Holáň, Vladimír (advisor) ; Stříž, Ilja (referee)
Regulatory T cells (Treg) are an important control mechanism within the Immune system (IS). Tregs prevent overactivation of effector T cells or autoreactive cells from invading organism-derived tissues. Treg are characterised by expression of surface molecules, CD4, CD25 and by an intracellular transcription factor forkhead box protein 3 (FoxP3). There are two basic populations of Treg, naturally occuring Treg (nTreg) developing in the thymus and induced Treg (iTreg) rising from CD4+ T cells in periphery, which are also precursors for T helper cells. In spite of an outgoing intensive research, there is still no clear clue which mechanisms are used by Treg to inhibit other effector cells. First in vitro experiments showed, that those mechanisms are of a contact dependent manner and do not use secreted molecules. But in vivo experiments showed the exact opposite. Those studies showed that secretory molecules, such as interleukin (IL)-10, IL-35 or transforming growth factor beta (TGF-β), are important in the effectory phase of Treg. Since the first experiments other distinct mechanisms of supression by Treg cells have been discovered. Those mechanisms seem to be important only in particular situations, particular cell assays or with using of specific experimental models. The reasons for this...
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The role of TNF-alpha and IL-10 in cardioprotective effect of chronic hypoxia
Chytilová, Anna ; Bezouška, Karel (advisor) ; Hloušková, Patricie (referee)
The aim of the present study was to find out whether adaption to chronic hypoxia affects the expresion of TNF-α and IL-10 in rat myocardium. TNF-α is a proinflammatory cytokine, which amplifies inflammatory reaction, while IL-10 has opposite antiinflammatory effect. We also measured concentration of nitrotyrosine as a marker of nitrosative stress. We used male Wistar rats divided into four groups: 1) normoxic controls; 2) exposed to continous normobaric hypoxia (10% O2) for three days or 3) for three weeks and 4) exposed to intermittent normobaric hypoxia (10% O2) for three weeks with one hour daily reoxygenation. Cytosolic and membrane proteins (cytosolic and particulate fractions) were obtained from the left ventricle, right ventricle and interventricular septum. Concentrations of TNF-α and IL-10 in both fractions were measured by ELISA. Continous hypoxia increased TNF-α production in particulate fractions from all ventricular parts and decreased the ratio of IL-10/TNF-α in particulate and cytosolic fractions. Intermittent hypoxia redistributed TNF-α from cytosol into the particulate fraction and prevented the drop of IL-10/TNF-α ratio in the cytosolic fraction. The highest concentration of nitrotyrosine was found in the particulate fraction from the right ventricle after three days of hypoxia....
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