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The use of immunoregulatory properties of mesenchymal stem cells/ and their therapeutic potential
Javorková, Eliška ; Krulová, Magdaléna (advisor) ; Stříž, Ilja (referee) ; Tučková, Ludmila (referee)
Mesenchymal stem cells (MSCs) have the potential to differentiate into various cell types, possess potent immunomodulatory properties and can influence various functions of immune cells. Since the immunomodulatory properties of MSCs can be modified by cytokines, we compered the effect of unstimulated MSCs and MSCs pretreated with interleukin (IL)-1, interferon (IFN)- , transforming growth factor (TGF)- and IL-10 on the development of regulatory T cells (Treg) and T helper 17 (Th17) cells in vitro and on the inflammatory environment in the eye. MSCs can produce significant levels of TGF- and IL-6. These cytokines represent the key factors that reciprocally regulate the development of naive T cells into Treg and Th17 cells. Unstimulated MSCs produce TGF- , but not IL-6, and the production of TGF- can be further enhanced by IL-10 or TGF- . In the presence of IL-1, MSCs secrete significant levels of IL-6, in addition to spontaneous production of TGF- . MSC producing TGF- induced preferentially expression of Foxp3 and activation of Treg lymphocytes, whereas MSCs supernatants containing TGF- together with IL-6 supported ROR t expression and development of Th17 cells. We demonstrated that MSCs and their products effectively control the development of Tregs and Th17 cells in a population of...
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The study of food allergy in patients and experimental model
Goliáš, Jaroslav ; Tučková, Ludmila (advisor) ; Holáň, Vladimír (referee) ; Prokešová, Ludmila (referee)
Food allergy belongs among the most frequent disorders and its incidence is continuously rising over the last two decades in the developed world. Although the methods used in the diagnostics of food allergies are high sensitive, they have low specificity, which is affected by a purity of used extracts. Therefore, it is important to develop new proteomic procedures for isolation of food allergens in the pure and the biologically active forms, thereby improving the diagnostics of food allergies. Another approach for studying allergies is using an experimental model, which can help us to clarify the mechanisms of allergic response and the acquired findings employ in prophylaxis or allergy treatment. In the first part, we have developed a new proteomic procedure for isolation of wheat allergens in the purified form. By this procedure, using Rotofor, HPLC and electrophoretic methods, we identified 27 potential wheat allergens, from which 7 were newly identified: endogenous α-amylase/subtilisin inhibitor, trypsin/α-amylase inhibitor CMX1/CMX3, TLP, XIP-1, β-glucosidase, class II chitinase, and 26 kDa endochitinase. Further, we showed that isolated allergens (α-amylase 0.19, LTP, TLP, and wheatwin) retained their biological activity and were capable to activate basophils (BAT). In the second part, we...
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Cytokine polarisation in immunopathological disorders
Durilová, Marianna ; Štechová, Kateřina (advisor) ; Tučková, Ludmila (referee) ; Slavčev, Antonij (referee)
6 Abstract Cytokines as primary modulators of immune system cells play a key role in their development, maintenance and activity of each cell population. Cytokine profile of peripheral blood mononuclear cells thus reflects the immunopathological events involved in pathogenesis of the disease. Focus of my thesis was cytokine dysbalance in several immunopathological disorders (type 1 diabetes mellitus, autoimmune thyroiditis, allergic colitis) with the aim to find distinctive cytokine profile of each disorder and to point out common features and differences in these disorders. Protein array and enzyme-linked immunosorbent assay (ELISA) method were used for analysis of cytokines in breast milk and cytokines produced in vitro by peripheral and cord blood mononuclear cells spontaneously and after stimulation by autoantigens. First immunological disorder to study was type 1 diabetes mellitus (T1D), where patients with T1D were studied (n = 10) as well as their first degree relatives (n = 9), with particular group of neonates whose one parent suffered from T1D (n = 52). In patients with T1D prediabetic phase was dominated by spontaneous and postimulatory production of Th1- associated cytokines (IFN-γ, TNF-β) which dropped at the time of T1D manifestation and remained suppressed for at least two following years....
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Food allergy to wheat flour proteins
Šotkovský, Petr ; Tučková, Ludmila (advisor) ; Panzner, Petr (referee) ; Prokešová, Ludmila (referee)
THESIS SUMMARY Food allergy is one of the frequent disorders and its incidence in paediatric as well as adult population is continuously rising, having doubled in the last two decades. Although wheat belongs to major food allergens and is a staple food in most diets, we have only little knowledge of wheat proteins causing IgE mediated hypersensitivity reaction. Diagnostic approaches of food allergy to wheat have a high sensitivity, but low specificity. Poor predictability and specificity may be associated with the insufficient purity of wheat extracts used in sIgE assays or with the lack of major allergens in these extracts. In the first step, we characterized 19 potential allergens recognised by IgE Abs of allergic patients, using proteomic techniques (1-DE, 2-DE, MALDI-TOF, QTOF and LCQDECA nLC-MS/MS ion trap technique). We identified these IgE-binding molecules such as: α-amylase inhibitors, β-amylase, profilin, serpin, β-D-glucan exohydrolase and 27K protein. To quantify sIgE in patient's sera we developed ELISA using the whole wheat extract and two commercially available α-amylase inhibitors. Second, we developed a procedure that allows isolation of wheat allergens from natural sources using Rotofor cell and HPLC. Twenty-seven potential wheat allergens have been successfully identified; of these, the...
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Disorders in regulation of immunity - allergy and autoimunne diseases
Kayserová, Jana ; Šedivá, Anna (advisor) ; Tučková, Ludmila (referee) ; Thon, Vojtěch (referee)
The pathology of immune system can lead to immune disorders. Immunodeficencies are caused by insufficient or missing immune response. On the other hand, allergies and autoimmune disorders represent a consequence of wrong control of the immune reaction and breakdown of an immune tolerance. Immunopathogenesis of allergic and autoimmune diseases are to some extent common to both immunopathologies; both represent harmful hypersensitive reaction to autoantigen or allergen and lead to the destruction of tissues and organs or to their dysfunction. Allergy and autoimmunity result from the combination of internal, mainly genetic, and external factors, such as infection. In this thesis, we focused on the mechanisms that lead to the disorders of regulation of immune reaction. We studied cohorts of patients with allergy or autoimmunity and we concentrated first on the genetic omponents that underlie both immun opathologies, furtheron mechanisms of innate immunity, particularly dendritic cells and finally on the adaptive immunity, mainly B cells and antibodies. One of our projects presented our experience with the therapy influencing B lymphocytes using monoclonal antibody against CD20 (rituximab). In summary, our studies present a complex view on immune reactions that contribute to allergic and autoimmune diseases. Our...
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