|
|
The role of dendritic cells in various pathological states
Sochorová, Klára ; Bartůňková, Jiřina (advisor) ; Tučková, Ludmila (referee) ; Stříž, Ilja (referee)
Dendritic cells (DCs) represent one of the most important components of the immune system. DCs are the most effective antigen presenting cells with a unique ability to stimulate naive T cells. They ensure the crosstalk between innate and adaptive immunity. They participate in anti-infectious and anti-tumor immune reaction as well as in the induction of tolerance. It is clear, that the defect in DC can be fatal for the organism. In our work we studied the biology of DCs and the disturbances of DCs function in pathological states. We analyzed DCs in patients with Bruton's tyrosin kinase deficiency, we compared the effect of vitamin D analogs, calcitriol and paricalcitol, on DCs and we set up the protocol of DC generation for the immunotherapy of ovarian cancer. In the study concerning btk deficiency and dendritic cell function we found profound impairment of IL-6 and TNF production in response to the stimulation by Toll-like receptor ligand 8. In the second part of our work we compared the effect of calcitriol and paricalcitol on DCs. Both drugs inhibited DCs maturation and decreased their ability to induce proliferation of antigen specific T cells. In the third part of our work we set up the protocol for the DCs, which are able to induce tumor specific immune responses. We optimized the form of tumor antigen...
|
|
|
Immunomodulatory properties of vitamin D3
Urbanová, Anna ; Stříž, Ilja (advisor) ; Zajícová, Alena (referee)
1 Abstract Vitamin D3 is important for keeping the right concetration of Ca2+ in plasma. Therefore it is essential for proper bone growth and development. Nevertheless, vitamin D3 has also a number of immunomodulating effects. Our thesis has been targeted on evaluation and comparison of vitamin D3 influence on expression of chosen surface markers (CD14, CD54, HLA-DR, CD16, CD36 and CD163) with THP-1 cells and monocytes gained from human peripheral blood. Other aims have been analysing the vitamin D3 influence on longevity of THP-1 cells and measuring the soluble CD14 and IL-8 production with THP-1 cells under the vitamin D3 influence. The cells have been stimulated with five different concentrations of vitamin D3 for the time 24, 48 and 72 hours. Higher used concetrations of vitamin D3, i.e. 100 nM and 1000 nM have increased the expression of CD14 with THP-1 cells in the time 48 and 72 hours of the stimulation time. With the monocytes from peripheral human blood the increase of the CD14 expression hasn't been remarkable from the physiological point of view. Together with the vitamin D3 concentration increase the sCD14 production with THP1 cells was considerably higher. The sCD14 was the highest in the time 72 hours after the stimulation with the highest used vitamin D3 concetration. The IL-8 quantity with...
|
|
|
The effect of polyketide antibiotics on signalling and functional activity of human monocytic cell line
Kopecká, Kristýna ; Stříž, Ilja (advisor) ; Kovářová, Jaromíra (referee)
Anti-inflammatory cytokines have an important role in the development of inflammatory reactions. If an acute inflammation turns into chronical it is very often a pathological phenomenon. Chronicle inflammations accompany a whole number of serious diseases with an unclear prognosis, such as some of the autoimmune diseases. Usually, the cause of these diseases is not quite clear and the treatment is mainly symptomatic with an effort to suppress the immunity system. For this purpose we use various immunosuppressant drugs, and biological treatment is used, too. Another possibility is to use bioactive secondary metabolites produced by various microorganisms. In this group there are for example macrolides antibiotics, and a big potential is also seen in the recently discovered polyketides. The objective of this work is to test the newly acquired secondary metabolites that were isolated in the Laboratory of Molecular Biology of Actinomycetes at the Czech Academy of Sciences. Tested were manumycin A, manumycin B, colabomycin E, asukamycin A, asukamycin D, β-rubromycin, deoxynybomycin. As comparative substances were used the macrolides antibiotics clarithromycin and azithromycin dehydrate, all of them commercial pharmaceuticals. These substances were tested on the monocytic line THP-1. Cells were stimulated...
|
|
|
ANCA-Associated Vasculitides : Complex Diagnostic Approach
Hrušková, Zdenka ; Posová, Helena (advisor) ; Stříž, Ilja (referee) ; Žák, Aleš (referee)
Objectives: The aim of this study was to assess cellular immunity parameters in patients with ANCA (Anti-Neutrophil Cytoplasmic Autoantibodies)-associated vasculitides (AAV) at different stages of the disease, with different treatment modalities, and with respect to the long-term prognosis of the patients. Methods: We examined 69 patients with AAV, 30 healthy individuals and 20 patients with chronic kidney disease. Using flow cytometry, the following markers were assessed in peripheral blood cells: surface molecules (CD4, CD8, CD3, CD19, CD80, CD86, HLA-DR, CD28, CXCR3, CCR5, CD30 and CRTH2) and intracellular cytokines (interferon gamma (IFN), tumor necrosis factor alpha (TNF), interleukin (IL)-2 and IL-4 in CD3+ T cells and IL-10 and IL-12 in monocytes). Results: Patients with AAV had decreased total number of lymphocytes, CD4+ cells, and CD4+CD45RA+ cells compared to healthy controls (p<0.001). Active patients had increased CD30 and CRTH2 expression (p<0.05). Increased CCR5 expression persisted in remission. Increased HLA-DR expression, expansion of CD28 subpopulation and increased IFN production were noted in remission but not in active disease. Patients in remission who developed a relapse during follow-up had significantly lower IL-10 production than those without relapse (p<0.01). Conclusions: Taken...
|
|
|
Immunomodulatory effects of macrolide antibiotics
Zemánková, Jana ; Stříž, Ilja (advisor) ; Krulová, Magdaléna (referee)
Macrolide antibiotics are well known not only for their antibacterial properties, but also for their recently discovered anti-inflammatory properties. They are able to significantly suppress destructive and in many cases life-threatening inflammation, an effect which is desired especially in chronic inflammatory diseases. The principle which their act is the modulation of the various components of the immune system. These effects are called "immunomodulatory" and can also include the effect on epithelial cells and their secretory activity, as well as the effect on pathogens which can colonize the airways and contibute to pathogenesis and the emergence of the chronic inflammatory respiratory diseases. This thesis summarizes the most important known mechanisms, by which macrolide antibiotics exert these immunomodulatory effects, and also notes examples of diseases whose treatment is the most clinically significant. Macrolide antibiotics posessing these uniqe anti-inflammatory properties are well tolerated and severe side-effects are rare. However, the most serious risk is the emergence of resistance and that is the main reason why this treatment can not be recommended without reservation. It is up to each doctor to consider the risks and benefits of the treatment in each individual patient.
|
|
|
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...
|
|
|
název v anglickém jazyce není uveden
Vítová, Andrea ; Filipec, Martin (advisor) ; Rozsíval, Pavel (referee) ; Stříž, Ilja (referee)
ZÁVĚR 1) Naše výsledky rozšířily pohled na obranné mechanismy zajišťované buňkami rohovkové tkáně o schopnost stromálních buněk rohovky selektivně potlačovat produkci některých (IL-4 a IL-10) protizánětlivých cytokinů. 2) Porovnáním účinnosti léčby imunosupresivními léky nebo mAb u dvou rizikových modelů myších příjemců jsme zjistili, že pro prevenci rejekce rohovkového alotransplantátů je efektivnější léčba mAb proti CD4+ buňkám než mAb proti CD8+ buňkám či imunosupresivními léky MMF, CsA nebo jejich kombinací. Léčba pomocí mAb byla efektivní pouze u příjemců s prevaskularizací rohovky. 3) Podávání séra s obsahem cytotoxických protilátek specifických proti antigenům dárce urychlilo nástup rejekce a významně zkrátilo přežívání alotransplantátů i xenotransplantátů rohovky. Z výsledků vyplývá, že na procesu rejekce aloi xenotransplantátů rohovky se může přímo podílet protilátkami zprostředkovaná imunitní odpověď. 4) Analýzou syngenních a xenogenních transplantátů rohovky ve zvolených obdobích po transplantaci jsme zjistili, že IFN-y a NO hraje podstatnou roli při rejekci rohovkových xenotransplantátů, zatímco exprese genů pro IL-2, IL-4 a IL-10 není pro rejekci vyžadována. Tato dizertační práce se zabývala molekulárními a buněčnými mechanismy probíhajícími ve zdravé a rejekované rohovkové tkáni. Pochopení...
|
|
| |
|
|
Dendritic cells in solid tumors of urogenital system
Minárik, Ivo ; Jarolím, Ladislav (advisor) ; Stříž, Ilja (referee) ; Soukup, Viktor (referee)
In the last 15 years we can observe many clinical studies which focus on recovery of immune reaction against tumor cells and possibly tumor elimination. At first it is necessary to understand the interaction of immune system with tumor cells so that effective vaccines are developed. In the first part of this work, I focus on the preparation of dendritic cell vaccine for patients with ovarian and prostate cancer. We show that dendritic cells can be differentiated from peripheral mononuclear cells in both tumors in vitro, we were able to pulse dendritic cells with apoptotic tumor cell, in case of ovarian cancer with autologous tumor cells, in case of prostate cancer with prostate cancer cell lines LNCaP and DU145. Pulsed mature dendritic cells expressed maturation marker CD83, costimulatory molecules CD80 a CD86 and produced significant cytokines. These dendritic cells also induced specific T lymphocyte response. In the next part of the work we focused on practical aspects of preparation of dendritic cell vaccine in the patient with prostate cancer and optimization of preparation in GMP conditions. The best medium for cultivation turned out to be Cell Gro. Maturation of dendritic cell with poly I:C led to the highest proliferation of specific T lymphocytes and at the same time to the lowest proliferation of...
|
|
| |
guest ::
