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NK cell involvement in the pathogenesis of autoimmune arthritis
Richter, Jan ; Fišerová, Anna (advisor) ; Hromadníková, Ilona (referee) ; Turánek, Jaroslav (referee)
Rheumatoid arthritis (RA) is a worldwide problem representing one of the most prevalent autoimmune diseases in the world. Despite the commonness of the disea- se, its pathogenesis has not been fully described. Immune cells ranging from antigen- presenting cells to T, B and NK cells playing various roles participate in the rheumatic process. In this work we concentrated on NK cells expressing a repertoire of activating and inhibitory receptors which influence their function in health and disease. We focused on the analysis of NK cell function and described its possible modulation by rheumatic autoantigens and multivalent glycodendrimers bearing 4 (GN4C) or 8 (GN8P) N-acetyl glucosamine moieties. The effect on NK cells and the glycosylation pathways was further studied in vitro. Finally, an in vivo study was performed on an animal model of RA - col- lagen-induced arthritis (CIA) to assess the effect of the compounds on clinical develop- ment of the disease and selected immune parameters. Comparison of NK cell cytotoxicity in patients suffering from RA, other inflam- matory diseases and healthy donors showed its impairment particularly in RA patients. Peripheral blood NK cells reacted to GN8P glycoconjugate by inhibition of their effector function in CD161 high-expressing samples. The MGAT5...
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Preparation and characterization of nanoliposomal carriers of hydrophobic cytostatics using nanofluidic mixing
Zelníčková, Jaroslava ; Mašek,, Josef (referee) ; Turánek, Jaroslav (advisor)
This diploma thesis is focused on preparation of liposome by relatively new method called nanofluidisation. This method allows the controlled preparation of small unilamellar liposomes in one step. In my thesis I was dealing with optimalization of liposomes preparation which carry hydrophobic cytostatics using this method. Cytotoxic effect of liposomes carrying hydrophobic cytostatics in vitro on cell lines A549 and MCF-7 was determined. In cytotoxicyty test I compared the effect of hydrophobic cytostatics (paclitaxel and derivates of vitamin E specifically alfa-Tos, alfa-TEA) that were incorporated into liposomes prepared via nanofluidisation method and lipid film hydration method. Moreover, a technology of lyophilisation in the presence of cryoprotectants for preparation of liposomes using the method of nanofluidisation was developed.
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Use of nanofluidic mixing for preparation of liposome carriers stained by gadolinium for contrast imaging by magnetic resonance (MRI)
Velínská, Kamila ; Mašek,, Josef (referee) ; Turánek, Jaroslav (advisor)
This diploma thesis focuses on the preparation of the liposomes, containing lipids with gadolinium, which are used for a contrast magnetic resonance imaging. The liposomes were prepared by the lipid film hydration followed by an extrusion and also by a new nanofluid mixing method on the NanoAssemblr Benchtop. The preparation technology has been optimized for parameters such as the composition of lipids, the flow rate ratio and total flow rate. The method of modification of the liposomes surface by gadolinium complexes has been developed. This method is using a conjugation reaction between the lipids containing cyanuric acid and Gd-DOTA. Prepared Gd-liposomes, which contain gadolinium, were complexly defined by the characterization techniques of DLS and NTA. The morphology of liposomes was observed by TEM and cryo-TEM. Methods for the determination of phospholipid content (Stewart test) and residual water in the lyophylisates of liposomes (Karl-Fischer titration) were used. Gadolinium in liposomal preparations was determined by ICP-OES. Using MR, the concept of gadolinium liposomes was verified and designed for MRI imaging of thrombi. The concept describing the mechanism of liposomes formation based on the experimentally proven existence of a phospholipid bilayer fragment has been developed. This concept is based on the experimentally proven existence of a phospholipid bilayer fragment.
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Immunoprotective character of polymer cytotoxic drugs
Šírová, Milada ; Říhová, Blanka (advisor) ; Turánek, Jaroslav (referee) ; Rabišková, Miloslava (referee)
18 I. INTRODUCTION Cancer is a serious health problem worldwide. In economically developed countries, it is a second most frequent cause of death after cardiovascular dis- eases, and the number of oncological patients continuously increases with the increasing age of population. The mainstay of cancer therapy is combination of surgery, radiation and chemotherapy. Whilst surgery and radiation are relatively precise and suitable to achieve a local control over the tumor, chemotherapy exerts a systemic ef- fect. These three modalities, when properly combined and sequenced, can cure a substantial number of hematological cancers and a smaller, but still significant subset of various solid tumors. Most cytostatic/cytotoxic drugs that are now in common use target the cells with high proliferation rate. The non-selective character of chemotherapy leads to increased toxicities towards normal rapidly proliferating cells. This means that the drugs have to be used at suboptimal doses, leading to development of (multi)drug resistance, metastatic disease and, eventually, to failure of the therapy. Innovative therapeutic strategies need to be developed in order to achieve better treatment outcome. For that purpose, several approaches are be- ing applied. First, sophisticated genomic and proteomic research could identify...
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NKR-P1C receptor-carbohydrate interaction contributes to antitumor immune response via activation of NK and B cells
Hulíková, Katarína ; Fišerová, Anna (advisor) ; Kročová, Zuzana (referee) ; Turánek, Jaroslav (referee)
Lectin - saccharide interactions and the involved receptors are currently intensively studied for their important role in antimicrobial as well as antitumor immunity. The major cell types participating in carbohydrate recognition are NK, NKT, and B cells. The differentiation of B lymphocytes could be induced by activated NK cells via direct intercellular contact and/or IFN-γ release. Our research is focused on NK cell receptors of C-type lectin-like family recognizing carbohydrate epitopes of glycoproteins. Synthetic glycoconjugates with terminal N- acetyl-D-glucosamines on polyamidoamine (GN8P) or calix[4]arene (GN4C) scaffold used in this study, exerted the highest binding affinity to activating isoforms of rodent NKR-P1 (A and C) receptor. The aim of the presented dissertation thesis was to elucidate, how Nkr-p1c gene divergence, between C57BL/6 (NK1.1-positive, NKR- P1CB6) and BALB/c (NK1.1-negative, NKR-P1CBALB/c) mouse strains, could affect NK cell activation and subsequent triggering of B lymphocyte effector functions using GN8P and GN4C as NKR-P1C prototype ligands. We demonstrated, that GN8P increased mRNA expression for NKR-P1C receptor, IFN-γ synthesis and lytic activity of NK cells, antigen-specific (anti-KLH, anti-DNP, and anti-B16F10) IgG (particularly IgG2a) formation, number of...
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In vitro and in vivo antimicrobial effect of lasioglossins on the Candida albicans
Kašperová, A. ; Turánek, J. ; Čeřovský, Václav ; Raška, M.
Lasioglossins represent a new group of amphipathic α-helical peptides with significant antimicrobial effect on the Candida albicans. This study examines the antifungal activity of two peptides LL-III and all D-LL-III as measured by the suppression of Candida proliferation and suppression of induced morphological differentiation both in in vitro and in vivo assays. In the in vitro Candida proliferation assay, the inhibitory effect of lasioglossins LL-III and all D-LL-III was more than 70% within 24 h and more than 84% after 48 h of incubation (final concentration of either peptide was 17.5 .mu.M). Delaying of blastoconidial transition to hyphae in vitro and tendency to suppress vaginal candidiasis in experimental mice were detected.
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