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Comparison of immunomodulatory properties of mesenchymal stem cells and Sertoli cells
Porubská, Bianka ; Krulová, Magdaléna (advisor) ; Komrsková, Kateřina (referee) ; Filipp, Dominik (referee)
Cell therapies are increasingly considered in preclinical studies and in the future of medicine. The main cell type investigated in this manner is mesenchymal stem cells (MSCs), because of their strong immunomodulatory properties. The efficacy of the therapy depends on various aspects, such as the viability and source of MSCs, the purity of the cell suspension and many more. There is a need for more tailored therapy and the use of cell type better fitting for the specific pathology. Sertoli cells (SCs) are deemed by some authors to be a kind of MSCs, namely because of their similar immunomodulatory properties. Because they reside in the seminiferous tubules in the testes, they are a promising candidate for the treatment of inflammatory pathologies of testicular tissue, such as bacterial infection-induced infertility. In vitro comparison of the ability of MSCs and SCs to differentiate into mesenchymal cell lineages such as osteocyte, chondrocyte, and adipocyte showed success in the case of SCs, providing evidence for their mesenchymal origin. The effect of MSCs or SCs on activated immune cells in vitro showed immunosuppression in both cases with distinct features. MSCs suppressed Th17 cell activation and IL-17 production by CD4+ T cells and SCs down-regulated TNFα and IL-2 production by these cells,...
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Anti-tumor and immunomodulatory effect of polymer conjugates based on HPMA carrying gemcitabine
Klusová, Natálie ; Beránek, Martin (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Natálie Klusová Supervisor: prof. PharmDr. Martin Beránek, Ph.D. Consultant: RNDr. Milada Šírová, Ph.D. Title of diploma thesis: Anti-tumor and immunomodulatory effect of polymer conjugates based on HPMA carrying gemcitabine The diploma thesis is focused on the anti-tumor and immunomodulatory effect of polymer conjugates based on HPMA carrying gemcitabine (Gem). We investigated the effects of four polymer conjugates (P-Gem1-P-Gem4), which had similar molecular weight and carried approximately the same amount of the drug. The only difference were the used spacers for Gem linkage. We used the following spacers: β-Alanin (P-Gem1); glycyl-phenylalanyl- leucyl-glycyl (P-Gem2); aminocaproic acid (P-Gem3); valeric acid (P-Gem4). Based on the assessed IC50 values for tumor cell lines 4T1, LL2, Panc02, MiaPaca2 and Panc1 the samples were divided into two groups: a) samples with quick rate of Gem release (P- Gem1, P-Gem2); b) samples with slow rate of Gem release (P-Gem3, P-Gem4). The in vivo stability of the conjugate affects systemic toxicity and anti-tumor activity, which was proven by the experiment performed on BALB/c mice bearing murine mammary carcinoma cells (4T1). We enrolled significant...
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Functional characterization of selected Kunitz proteins of Eudiplozoon nipponicum
Tymich, Alexandr ; Mikeš, Libor (advisor) ; Kašný, Martin (referee)
Proteins containing the Kunitz domain are mostly 6-10 kDa inhibitors of serine proteases, but in exceptional cases they can also inhibit cysteine and aspartic proteases. The main characteristic is the presence of six cysteine residues forming three disulfide bridges creating a typical active loop, which is complementary to the active site of various proteases. The specificity of this binding is largely determined by the amino acid in the P1 position. Their functions include the regulation of a number of physiological events based on proteolysis, e.g. the blood coagulation cascade or immune reactions. However, due to their nature, they have also become a powerful tool for parasitic organisms to interact with their host, where they again target proteases involved in the host's physiological events and thus allow the parasite to survive the interaction with the host. Until recently, representatives of the class Monogenea were a neglected group from the point of view of molecular parasite-host interactions, and only a few works were devoted to their biochemistry and the description of biologically active molecules. In this work, I focused on two selected Kunitz proteins in Eudiplozoon nipponicum, a blood-sucking ectoparasite from the Monogenea class, which has become a fairly common parasite of common...
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Anti-tumor and immunomodulatory effect of polymer conjugates based on HPMA carrying gemcitabine
Klusová, Natálie ; Beránek, Martin (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Natálie Klusová Supervisor: prof. PharmDr. Martin Beránek, Ph.D. Consultant: RNDr. Milada Šírová, Ph.D. Title of diploma thesis: Anti-tumor and immunomodulatory effect of polymer conjugates based on HPMA carrying gemcitabine The diploma thesis is focused on the anti-tumor and immunomodulatory effect of polymer conjugates based on HPMA carrying gemcitabine (Gem). We investigated the effects of four polymer conjugates (P-Gem1-P-Gem4), which had similar molecular weight and carried approximately the same amount of the drug. The only difference were the used spacers for Gem linkage. We used the following spacers: β-Alanin (P-Gem1); glycyl-phenylalanyl- leucyl-glycyl (P-Gem2); aminocaproic acid (P-Gem3); valeric acid (P-Gem4). Based on the assessed IC50 values for tumor cell lines 4T1, LL2, Panc02, MiaPaca2 and Panc1 the samples were divided into two groups: a) samples with quick rate of Gem release (P- Gem1, P-Gem2); b) samples with slow rate of Gem release (P-Gem3, P-Gem4). The in vivo stability of the conjugate affects systemic toxicity and anti-tumor activity, which was proven by the experiment performed on BALB/c mice bearing murine mammary carcinoma cells (4T1). We enrolled significant...
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Immunomodulatory effect of tumor targeted polymer drugs
Mervartová, Ivana ; Šírová, Milada (advisor) ; Palich Fučíková, Jitka (referee)
5 Abstract Myeloid-derived suppressor cells (MDSC) are a very heterogeneous population of immature, activated myeloid progenitors of neutrophils, monocytes/macrophages and dendritic cells that have not differentiated into mature forms. A common feature of these cells is the ability to suppress immune responses of T cells, NK cells, and dendritic cells. It is known that MDSC accumulate under various pathological conditions, such as chronic inflammation or cancer. In breast cancer patients, the highest MDSC counts correlate with the occurrence of metastatic foci in lung tissue. The suppressive effects of MDSCs are associated with resistance to chemotherapy, reduced effectiveness of immunotherapy and overall poor prognosis of the disease. Therefore, many studies focus on MDSC. One possibility is the differentiation of MDSC into mature populations that lose their suppressive phenotype. In this work, we focused on modulation of MDSC activity by all-trans retinoic acid (ATRA), bound to a polymer conjugate based on N-(2-hydroxypropyl)methacrylamide (HPMA). ATRA is used in clinical practice for the treatment of acute promyelocytic leukemia, where the mechanism of action is the differentiation of pathological cells into more mature forms and thus the cessation of their proliferation. The binding of ATRA to the...
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Rozdíly v charakteristických vlastnostech a funkcích invariantních NKT buněk zdravých dárců
DOSTÁLOVÁ, Karolína
iNKT cells are a subset of T cells, they express the same phenotypic markers usually found on T and natural killer (NK) cells. They comprise a lineage of CD1d-restricted glycolipid-reactive T lymphocytes which have an important role in imunity. They play critical role in the regulation of many different types of immune responses, ranging from self-tolerance and development of autoimmunity to responses to pathogens and tumors. These cells are gaining increasing importance in health care, particularly in hematooncology. In this study we examined the essential features and functions of Invariant Natural killer T cell (iNKT cells).iNKT cells were isolated from mononuclear cells (MNCs) of healthy donors. Then they were cultured under appropriate conditions in vitro. After reaching sufficient concentration and purity the iNKT cells were used for cytotoxic and immunomodulatory assays and immunophenotype determination. Our results showed that iNKT cells have varying qualities across donors and that they have a heterogeneous immunophenotype, cytotoxic and immunomodulatory potential.
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Immunomodulatory potential of Sertoli cell progenitors in Xenopus tadpoles during the healing of amputated tail
Mertová, Irem ; Krylov, Vladimír (advisor) ; Procházka, Jan (referee)
A cell culture of common Sertoli and peritubular myoid cells progenitors derived from the testes of male X. tropicalis, called XtiSC, was established in the Laboratory of Developmental Biology. XtiSCs exhibit similar properties as more well-known mesenchymal stem cells that are used in cell therapy for their immunomodulatory and proregenerative properties. Microinjection of XtiSC into the dorsal vein of the tail of the tadpoles has increased macrophage numbers 7 days after tail amputation (dpa), both in controls and in tadpoles after depletion of macrophages by the application of clodrosomes. Macrophage depletion also reduces the migratory ability of XtiSC to the site of tail amputation. Macrophage depletion also led to a reduction in the number of satellite cells 1 dpa. On the seventh day after tail amputation and XtiSC injection, there was a significant increase in their number compared to the control group without XtiSC injection. Using DAF-2DA probe, nitric oxide production was confirmed by injected XtiSC and at the injury site. For future research of the proliferation, differentiation and migration of satellite cells in vivo, a vector expressing EGFP under endogenous Pax7 promoter and Katushka-RFP under gama-crystalline promoter was created and integrated into the X. tropicalis genome using...
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Immunomodulatory properties of mesenchymal stem cells - use in therapy
Pavlíková, Michaela ; Krulová, Magdaléna (advisor) ; Stříž, Ilja (referee)
Mesenchymal stem cells (MSC) are extensively studied mainly due to their feasible clinical application. Therapeutic potential of MSC consists not only of the ability to differentiate into mesenchymal cells, ectodermal and endodermal cell lines, but primarily in their immunomodulatory functions. Due to their effect on immune cells, MSC promote the shift of the inflammatory immune response to antiinflammatory. The ability to suppress inflammation, together with their differentiation potential and antiapoptotic potential on the surrounding cells makes MSC a promising tool for treating serious diseases. This work discusses the effect of MSC on the individual cells of the immune system. It focuses on the description of the effect of MSC in four model cases. These are an experimental autoimmune encephalomyelitis, myocardial infarction, diabetes mellitus and skin graft transplantation. The knowledge of the mechanisms of the interactions between MSC and the cells of the immune system, together with the understanding the effect of specific conditions on MSC is essential for their use in clinical therapy. Keywords: mesenchymal stem cells, immunomodulation, autoimmune diseases, transplantation
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NK cells and their receptors in immune regulation - possible targets for immunomodulation
Svoboda, Jan ; Fišerová, Anna (advisor) ; Pěknicová, Jana (referee) ; Kročová, Zuzana (referee)
(english) Natural Killers - NK cells play an important role in immune surveilance and regulation either by direct cytotoxicity towards infected, transformed or otherwise damaged cells, or by production of cytokines and chemokines. The resulting response of NK cells is given by the sum of stimulating and inhibiting signals, tranduced by a wide array of receptors. Killer Ig-like receptors KIR2DL4 and LILRB1, which recognize self HLA-G molecules in pregnancy, as well as NKR-P1 receptors, which differ in the number of isotypes, are species-dependent and reduced during phylogenesis. NKG2D, reacting to stress-inducible proteins, and adenosine receptors (AR), which supress the inflamatory reaction, remain evolutionary conserved. The aim of this work was to study the involvement of NK cells and their receptors in several immune disorders and in various species, to provide new insights into their function and posisible immune modulation. We have shown here, that the choice of species in the study of NK cell effector functions may be crucial in some cases. The reaction to glycans, using synthetic GlcNAc-terminated glycomimetics GN8P, exerted opposing effects on NK cell function in humans and C57Bl/6 mice. In humans, the glycomimetic decreased cytotoxic activity of high NKR-P1A expressing NK cells, while in...
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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.
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