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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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Cytokine/anti-cytokine mAb complexes and their biological activity
Hnízdilová, Tereza ; Kovář, Marek (advisor) ; Grobárová, Valéria (referee)
biologická aktivita některých cytokinů může být paradoxně zvýšena tvorbou komplexu některými svými monoklonálními protilátkami (mAb) rozpoznávajícími tento cytokin. Prodloužení poločasu eliminace z 2 mAb komplexy, neboť v použitém klonu mAb dochází k selektivní stimulaci buď CD25 2/S4B6 komplex) buněk. Díky výrazné stimulaci NK buněk a paměťových CD8 lymfocytů a pouze mírné stimulaci Treg buněk by IL nahradit konvenční IL vysoce selektivní stimulací Treg buněk naopak mohly uplatnit při léčbě autoimunitních onemocnění a transplantacích. Potenciálně klinicky využitelné jsou dále IL stimulaci žírných buněk, IL plazmatickými buňkami či IL proliferaci a přežívání T lymfocytů. Imunokomplexy mohou být tvořeny také cytokinem a jeho 15Rα komplexy představují generaci IL onistů, které mohou být díky stimulaci expanze NK buněk a paměťových CD8 T lymfocytů využity jako Klíčová slova 15Rα komplexy
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Therapeutic Nanoparticles and Immunotoxicity
Lehutová, Daniela ; Vannucci, Luca Ernesto (advisor) ; Grobárová, Valéria (referee)
Nanočastice sú submikroskopické štruktúry vyhotovené z rôznych materiálov s obrovským terapeutickým potenciálom. Rovnako predstavujú nový nástroj pre lepšie cielenie a podávanie liečiv do jednotlivých tkanív. Bolo vynaložených mnoho pokusov pre vyvinutie terapeutík, ktoré budú lepšie kompatibilné v organizme a zároveň by zlepšili účinnosť moderných liekov. Na rozdiel od bežne používaných spôsobov pre podávanie liečiv, enkapsulovaná forma predstavuje obrovskú výhodu pri znižovaní vedľajších účinkov oproti bežne používaných liekoch. V tejto práci sú porovnávané výhody použitia nanočastíc spolu s ich možným rizikom, najmä na ich munitný systém. Vzhľadom k tomu, že toxicita sa líši v závislosti na chemicko fyzikálnych vlastnostiach a tkanivách, v ktorých sa akumulovali, musí byť zvolená vhodná stratégia, aby sa predišlo možných nežiaducim účinkom. Z tohto dôvodu je potrebné zvážiť preventívne opatrenia a správne upraviť dané nanočástice predtým, než budú použité v organizme, rovnako ako spôsobená odpoveď organizmu a jej dôsledok musia byť vhodne monitorované. Zvýšená pozornosť musí byť venovaná príprave nanočastíc, aby sa predišlo prípadnej kontaminácii a nebezpečenstvu pre laboratórny personál. Kľúčové slová nanočástice, podávanie liečiv, imunita, toxicita, teranostika
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The role of bacterium Escherichia coli in the tumorigenesis of colorectal carcinoma
Čurnová, Lenka ; Klimešová, Klára (advisor) ; Grobárová, Valéria (referee)
Colorectal carcinoma is a severe disease of colon. It belongs to the cancers with the highest incidence and also high mortality. In the process of tumorigenesis, there are applied various mechanisms, mainly DNA damage and subsequent reparation and inflammation. Gut microbiota plays an important role in development of the colorectal cancer influencing cancer microenvironment. Microbiota triggers inflammatory response or produces different toxins. The diploma thesis was aimed on the relation between the presence of cyclomodulin genes in the genome of individual strains of bacterium Escherichia coli and their genotoxic features. To follow direct influence of epithelial cells by microbiota, we used in vitro model. We chose E. coli as a model microorganism because it is common bacterium of human gut, moreover, as facultative anaerob it is easily cultivated. We used six strains of E. coli with different relation to the host organism including probiotic, comensal and pathobiont. In probiotic strains of E. coli (Nissle 1917 and O83), we detected less genes for cyclomodulins than in other strains. We did not observed significant differences in genotoxic features of the strains. Also, we did not detect any changes in viability, proliferation, activation of repair mechanisms or p53 fosforylation caused by...
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Mechanisms of immunosuppressive drug-mediated effect on mesenchymal stem cell properties
Vacková, Julie ; Krulová, Magdaléna (advisor) ; Grobárová, Valéria (referee)
Mesenchymal stem cells (MSC) defined as multipotent, nonhematopoietic stem cells have been shown to possess various immunosuppressive properties. Thus they can be used to attenuate transplant rejection and also for treatment of autoimmune diseases. But simultaneously with MSC patients take immunosuppressive drugs and there is no evidence how this medication affect MSC. The goal of this study is to elucidate how frequently used immunosuppressive drugs cyclosporineA, mycophenolate mofetil, rapamycin, dexamethasone and prednisone influence immune-related parameters of mice and human MSC. Here we show that MSC from various sources are affected differentialy after short-term exposure of the tested immunosuppressants. Only cyclosporine A does not change immune-related parameters of mice MSC in the comparison to other immunosuppressants. However, cyclosporine A, mycophenolate mofetil and rapamycin enhance human MSC expression of TSG-6, PD-L1 and TGF-β which are involved in inhibition of lymphocyte proliferation and effector function, inhibition of dendritic cell maturation and in support of tolerogenic phenotype of macrophages. Although glucocorticoid drugs promote survival of human MSC and expression of Fas-L they reduce expression of molecules that mediate immunosuppression. In this respect, the best...
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Biological activity of IL-2/anti-IL-2 mAb immunocomplexes in vivo and their terapeutical potential
Hnilicová, Šárka ; Kovář, Marek (advisor) ; Grobárová, Valéria (referee)
IL-2 belongs to the family of c cytokines (IL-2, 4, 7, 9, 15, and 21) which are key regulators of lymphocyte homeostasis and function. They have the potential to promote lymphocyte proliferation and survival and thus overall enhance dominantly adaptive immune response. IL-2 is an autocrine/paracrine soluble factor produced mainly by activated T cells. Interestingly, the in vivo biological activity of IL-2 can be dramatically increased through complexing with certain anti-IL-2 mAbs and such IL-2/anti-IL-2 mAbs immunocomplexes selectively stimulate proliferation of distinct population of immune cells, depending on the clone of anti-IL-2 mAb used. IL-2/S4B6 mAb immunocomplexes are highly stimulatory for CD122high populations (memory CD8+ T and NK cells) and intermediately also for CD25+ populations (Treg and activated T cells), while IL-2/JES6-1 mAb immunocomplexes enormously expand solely CD25+ cells. Thus, IL-2 immunocomplexes possess a broad spectrum of potential therapeutic applications like tumor immunotherapy, vaccination, autoimmune diseases or transplantology.
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Molecular mechanism of MST1 kinase activation in cancer cells
Smetanová, Jitka ; Vališ, Karel (advisor) ; Grobárová, Valéria (referee)
MST1 kinase is an internal part of the Hippo signal pathway. The Hippo pathway is an evolutionary conserved regulator of tissue and organ growth and affects proliferation and apoptosis. Active MST1 kinase phosphorylates YAP and TAZ oncoproteins, which regulate the activity of transcription factors in their unphosphorylated state, including TEAD and SMAD. Furthermore active MST1 kinase phosphorylates FOXO transcription factors and induces their translocation into the cell nucleus. Finally the activation of MST1 kinase leads to cell apoptosis or halt cell cycle in G1 phase. Activation of MST1 protein depends on its auto-phosphorylation and cleavage. Recently, there are several articles which take interest in the issue of activation of MST1. Some of them describe the activation of MST1 by the effector caspase-3 and -7, on the other hand the latest articles argue that MST1 kinase itself is responsible for the activation of caspases. The molecular mechanism of MST1 kinase activation was studied in this bachelor thesis. We used the biologically active compounds GDC-0941 for the activation of MST1 protein. The activity of caspase was inhibited by specific inhibitor Z-DEVD. Using electrophoresis and Western blot it was demonstrated that MST1 is active in the case when caspases are inhibited. This fact...
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Immunoprotective properties of Sertoli cells
Porubská, Bianka ; Krulová, Magdaléna (advisor) ; Grobárová, Valéria (referee)
Sertoli cells (SCs) are somatic cells located in the male reproductive organs- testes. Everyday, understanding of their function and their role in spermatogenesis becomes better elucidated and there is no doubt that reproduction and continuity of the kind would be impaired in the absence of SCs. SCs are not only able to influence spermatogenesis they also significantly modulate immune system. Both cell and humoral component of immune system are affected after SCs application and SCs thus protect not only themselves, but also other co-transplanted cells. Modulation of response of innate and adaptive component of immune system may play a key role in the treatment of several diseases. Powered by TCPDF (www.tcpdf.org)
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Mechanism of anti-cancer activity induced by sesquiterpene lactones.
Filandr, František ; Novák, Petr (advisor) ; Grobárová, Valéria (referee)
One way to stop tumor growth in organism is to induce differentiation, apoptosis or necrosis of tumor cells. A class of chemicals known to induce apoptosis and inhibit proliferation in leukemia cells are sesquiterpene lactones. One of these lactones is cnicin, a bitter tonic used in liqueurs, found in the plant Cnicus benedictus. The mechanism of this inhibition, is not fully understood but certain signaling pathways are suspected, mainly the recently discovered Hippo signaling pathway which controls the organ size and apoptosis in mammalian cells. The core kinase of this signaling pathway is MST1/2 protein and its activation by sesquiterpene lactone cnicin resulting in cleavage of its active N-terminal domain is observed in this work. Also, the effect of cnicin on down-regulating main oncoprotein deregulated in leukemia cells C-MYC is studied. In addition the results of q-PCR also show significant down-regulation of anti-apoptotic BCL2 and MCL1 genes and cMYC oncogene. (In Czech)
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Lectin receptor-ligand interaction important in experimental tumor therapy
Grobárová, Valéria ; Černý, Jan (advisor) ; Filipp, Dominik (referee) ; Krulová, Magdaléna (referee)
Lectin-saccharide interactions are involved in many biological processes essential for the survival and proper function of multicellular organisms. C-type lectin-like receptors, predominantly expressed by cells of the innate immune system, recognize saccharide structures on microbes and also aberrant glycosylation pattern of cancer cells. The NKR-P1 receptor family was among the first natural killer (NK) receptor families that were identified, however ligands for some of members remain still elusive. Recently, publications describing N-acetylglucosamine-terminated oligosaccharide structures as possible ligands for NKR-P1 receptor have been subjects for correction/retractions after investigation of the Ethical Committee of the Institute of Microbiology, ASCR, v. v. i. and Charles University in Prague. Re-evaluation of glycodendrimer effect, particularly effect of N-acetyl-D-glucosamine octabranched dendrimer on polyamidoamine scaffold (GN8P), revealed mostly indirect role of NK cells on modulation of immune responses. Properly folded soluble recombinant rat NKR-P1A and mouse NKR-P1C lack binding activity to neoglycoproteins modified with GlcNAc-terminated structures. Moreover, new possible target cell populations (NKT cells and macrophages) for saccharide binding were identified.
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