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Immune response of different subpopulations of dendritic cells to probiotic strain of E. coli O83:K24:H31
Gorelová, Miroslava ; Hrdý, Jiří (advisor) ; Grobárová, Valéria (referee)
Allergy, as one of the worldwide most frequent pathologies, belongs to illnesses with constantly growing incidence among young children. Identification of prognostic markers pointing to increased risk of allergy development, allows introduction of early preventive measures. Probiotic supplementation could be one the preventive measure. It has been shown that introduction of selected probiotic strains or mixtures can prevent development of allergy. In this diploma thesis, the capacity of probiotic strain Escherichia coli O83:K24:H31 (E. coli O83) to support maturation of dendritic cells and polarization of immune responses was tested. Introduction of this probiotic vaccine called Colinfant Newborn appears to be suitable preventive measure, lowering allergy incidence in children with predisposition to development of allergy. The capacity of E. coli O83 to support maturation of the two main subpopulations of dendritic cells (myeloid dendritic cells - mDC and plasmacytoid dendritic cells - pDC) in cord blood of newborns of healthy mothers (children with relatively low risk for allergy development) and allergic mothers (children with relatively high risk for allergy development) was measured by flow cytometry. The presence of cytokines and transcription factors characteristic for particular...
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Different characteristics of cord blood cells of newborns of healthy and allergic mothers
Vlasáková, Kateřina ; Hrdý, Jiří (advisor) ; Hájková, Michaela (referee)
The prevalence of allergy is increasing and it is becoming a serious problem not on- ly in medicine, but also in social and economic terms. The most effective way to minimize the development of allergic diseases is preventive measures. In recent years, many studies have attempted to confirm or rebut the hypothesis that early administration of probiotic bacteria to newborns and pregnant women before birth could have preventive effects on the development of allergy. In the Czech Republic, the probiotic strain Escherichia coli O83:K24:H31 (EC O83), being registered with the State Health Institute for Drug Control under the name Colinfant Newborn, has long been used to prevent allergies and paediatri- cians have long been known and used it against various diarrhoea. The aim of this work was to elucidate the effect of EC O83 on CBMC (cord blood mononuclear cells) and to compare the ability of CBMC of healthy mothers (children with a relatively low risk of developing allergic disease) and allergic mothers (children at high risk of developing allergies) to form cytokines in response to EC O83 stimulation. Phytohemagglutinin was used as a positive control, Escherichia coli Nissle 1917 was used as a reference probiotic strain, which is much more known abroad than EC O83. Cytokine production was detected by...
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The role of myeloid dendritic cells in allergic diseases
Navrátilová, Adéla ; Hrdý, Jiří (advisor) ; Hájková, Michaela (referee)
Dendritic cells (DC) of hematopoietic origin can be distinguished into cells originating from the myeloid (mDC) or lymphoid lineage. DCs of both branches are able to uptake the antigens, present their samples to T-lymphocytes and polarize the immune response. Lymphoid line DCs are specialized in fighting viruses and they are an important source of interferon alpha. MDCs, which engage antigens as one of the first cells, play an important role in allergies, which are a current problem. Allergy is defined as an inadequate immune response to environmentally nonharmful antigens, and asthma, allergic rhinitis, atopic dermatitis and food allergy are among the most common allergic diseases. This work focuses on the role of mDC in the pathogenesis of allergic diseases and their possible use in treatment. Furthermore, this paper describes the differences between healthy and allergic individuals in terms of functional changes of mDC and differences between mature and immature immune system. Key words: dendritic cells, allergic diseases, cytokines, newborn immune system
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The role of innate lymphoid cells in influenza virus infection
Mouyabi, Flaviancia ; Hrdý, Jiří (advisor) ; Hájková, Michaela (referee)
Innate lymphoid cells (ILCs) are recently discovered group of innate immune cells. They do not have antigen-specific receptors but they can be activated by cytokines similarly to T lymphocytes. ILCs have a crucial role in the regulation of inflammation, tissue repair, containment of commensals, anti-infection immunity and regulation of tissue homeostasis. The presence of mouse and human ILCs can be detected in the lung during and after influenza virus infection when ILC contribute to the restoration of damaged lung parenchyma. ILCs directly or indirectly provide protection against viral infections by secretion of various cytokines and co-operation with other cells (e.g. T cells, macrophages). Overall, lung ILCs are important in immune responses and tissue homeostasis, but further studies on this topic are needed to fully understand their role. The aim of this thesis was to specifically characterize these cells, focus on their function in the lung, and describe their role in the course of influenza virus infection.
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Targeted differentiation and transdifferentiation of stem cells and their therapeutic application
Chudíčková, Milada ; Holáň, Vladimír (advisor) ; Bernášková, Klára (referee) ; Hrdý, Jiří (referee)
Mesenchymal stem cells (MSCs) naturally differentiate into cells of tissues of mesodermal lineage: cartilage, bone or adipose tissue. As a result of specific environmental stimuli, MSC are able to transdifferentiate into cells of endodermal or ectodermal lineage. Also, MSCs are able to regulate the inflammatory processes and to support healing and regeneration. These properties make MSCs suitable in cell-based therapy and tissue engineering. Characteristics of MSCs (for example differentiation and proliferative potential and cytokine secretion profile) can vary slightly depending on their origin. These differences can be further amplified by the effects of specific environments. Thus, to obtain maximal benefit, it is important to select MSCs optimal for a particular environment. The main goal of this thesis was to design in vitro protocol for transdifferentiation of MSCs into neuron-like cells. For this application, the adipose tissue-derived MSCs seemed to be optimal, due to their higher production of basic fibroblast growth factor, one of the important factors in neural development. The resulting cells acquired typical neuron-like morphology, expressed genes for neuron-specific markers and produced neuron-specific proteins. Further, the resulting cells showed immunomodulatory properties similar...
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Molecular mechanisms of the resistence of breast cancer cells to taxanes: the role of ABC transporters
Kopperová, Dana ; Kovář, Jan (advisor) ; Hrdý, Jiří (referee)
Resistance to chemotherapeutics is a widespread phenomenon in cancer cells that may counteract the successful therapy of many patients. In resistant cells, higher level of ABC transporters, among others, often can be detected. This high level of ABC transporters represents a suspected mechanism of acquired cancer resistance. We studied the molecular mechanism of resistance to taxanes in cancer cells using SK-BR-3 and MCF-7 breast cancer cell lines. We analyzed the effect of paclitaxel on apoptosis induction in the originally sensitive cells of these lines as compared to their counterpart resistant cells, developed by gradual adaptation to paclitaxel. In resistant cells of the SK-BR-3 and MCF-7 lines, we did not detected ongoing induction of apoptosis but we did detect significantly increased expression of ABCB1 transporter after paclitaxel application. By silencing the expression of the transport via employment of small interfering RNA (siRNA), we tested the role of the ABCB1 transporter in cells resistant to paclitaxel. We found that resistant cells with silenced expression of the ABCB1 transporter had a statistically significant increase of sensitivity to paclitaxel as compared to control resistant cells with high expression of this transporter. Along with increased sensitivity, we demonstrated...
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Immunologic Characteristics of Cord Blood in Children with Increased Risk of Allergy Development Preventive Use of Probiotics
Hrdý, Jiří ; Prokešová, Ludmila (advisor) ; Tlaskalová - Hogenová, Helena (referee) ; Ulčová-Gallová, Zdeňka (referee)
Allergy is one of the most common diseases. Identification of early prognostic markers pointing to an increased risk of allergy development is therefore of increasing importance. Cord blood represents an easily attainable clinical material for searching for prognostic markers signalizing future allergy development. Proportions of Th1 cytokines, Th2 cytokines and regulatory cytokines were tested in cord blood of children of allergic mothers (children in relatively high risk of allergy development) in comparison with cord blood of children of healthy mothers (low risk children). Also the activities of lymphocytes, dendritic cells (DC) and regulatory cells (Tregs) were compared in children of healthy and allergic mothers. The generally increased activity of both in vitro stimulated and non-stimulated mononuclear cord blood leukocytes was proved in children of allergic mothers in comparison with low risk children. The increased activity of DC of high risk children was detectable only after polyclonal stimulation. Significantly less pronounced functional properties of cord blood Tregs were found in children of allergic mothers when compared with children of healthy mothers. The increased reactivity of lymphocytes and DC together with the decreased activity of Tregs can support an easier...
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Comparison of cytokine production by leukocytes from newborns of healthy and allergic mothers
Dusilová, Adéla ; Hrdý, Jiří (advisor) ; Janatková, Ivana (referee)
The increasing incidence of children suffering from allergic diseases could be caused by sensitization of immature immune system during the intrauterine development. Several important scientific papers have demonstrated the ability of cord blood cells to respond by elevated proliferation activity after stimulation by common allergens. Following these findings, present study follows the production of cytokines which play a role in the pro- and anti-allergenic tuning of the immune system. Umbilical cord blood cells were stimulated with polyclonal activators (phytohaemagglutinin) and common allergens (ovalbumin, timothy grass, birch, mite). Subsequently, cytokine production was monitored using selected methods that reflect different stages of cell activation - at the level of mRNA by quantitative real time PCR (qRT-PCR), by flow cytometry detection of the presence of intracellular cytokines in different cell subpopulations and by ELISA measurement of cytokines in CBMC culture supernatants. The results obtained point to a very weak ability of these common allergens (timothy grass, birch, mite, ovalbumin) to stimulate CBMC to produce cytokines observed by all of these methodological procedures. Although we did not observe significant differences in CBMC cytokine production (IL-2, IL-4, IL-10, IL-12,...
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Comparison of Immune System of Newborns and Adults
Dusilová, Adéla ; Zajícová, Alena (referee) ; Hrdý, Jiří (advisor)
In general, it is possible to characterize neonatal immune system (IS) as immature in comparison to adult IS. From a clinical point of view, newborns show an increased susceptibility to infections. Breastfeeding can contribute to the descent incidence of illnesses, because it supplies the intestinal mucosal system with antibodies of the mother`s origin, important nutrients and other immunoregulatory components. Breast milk compensates decreased newborn's capacity to produce immunoglobulins- especially IgA, that concentration reaches adult levels in two years, but even later (to the pubescent period). Other classes of antibodies are found in cord blood only sporadically except IgG, which is transferred transplacentary. Reduced ability of B lymphocytes to produce antibodies is caused by insufficient expression of surface costimulatory signals of Th2 cells. T lymphocytes are not able to react properly to low doses of stimulators (polyclonal activators - phytoid lectins: ConA or PHA), which bind to T cell receptors in complex with CD3 and proliferate in a response to anti-CD3 monoclonal antibodies. Most of the cord blood T lymphocytes display "naive" phenotype CD45RA. During intrauterine development, neonatal IS is in contact with mother IS and because a pro-inflammatory Th1 response could lead to...
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