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Methods for Leishmania parasite detection and quantificatio as a tool for study of the pathogen-vector-host interactions
Kobets, Tetyana ; Lipoldová, Marie (advisor) ; Kodym, Petr (referee) ; Kopecký, Jan (referee)
Methods for Leishmania parasite detection and quantification as a tool for study of the pathogen-vector-host interactions Leishmaniasis in human is caused by total 21 species of the intracellular protozoan parasite Leishmania, which are transmitted by about 30 species of phlebotomine sand flies. Besides human, Leishmania can infect a number of vertebrate hosts. The major host cell is the macrophage, in which parasites multiply, eventually rupturing the cell and spreading to uninfected cells. Infected monocytes and macrophages circulating in the peripheral blood are thought to be carriers of the parasite to distal sites. Depending on the infected sites of the body, there are three forms of leishmaniasis: cutaneous, mucocutaneous and visceral. Leishmaniasis is a disease for which we still lack effective, affordable and easy to use drugs. In addition, surveillance and control are also neglected. This thesis summarizes the results of several projects using different approaches for parasite load measurement in the mouse model of leishmaniasis, including two methods that were developed and optimized in our laboratory. Detection and quantification of pathogens belongs to the major topics of the research of various infectious diseases. This parameter is necessary for confirmation of the diagnosis, characterization...
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Pre-clinical model of acute promyelocytic leukemia:/study of the anti-leukemic effect induced by ATRA and DNA vaccination
Pokorná, Kateřina ; Holáň, Vladimír (advisor) ; Stöckbauer, Petr (referee) ; Degos, Laurent (referee)
DOCTORAL THESIS 2012 POKORNA Abstract We have used a well characterized transplantable transgenic mouse model which mimics human acute promyelocytic leukemia (APL), both in its biological characteristics and its response to conventional therapeutic drugs. The aim of our study was to better characterize the efficacy of the combined treatment and to determine molecular markers of clinical outcome. We established a minimal residual disease monitoring based on the high sensitivity of detection of PML-RAR transcripts by polymerase chain reaction (PCR) technology in APL mice. We showed that oncogene-specific PCR-based assays allow, like in patients, the diagnosis, follow-up and prediction of disease evolution. Furthermore, PCR assay was used to assess various tissues and organs for the presence of PML-RAR-positive cells in minimal residual disease free long-term survivors. As expected, majority of mice had no measurable tissue level of PML-RAR demonstrating the efficacy of immunotherapy. However, tracking the oncogene-positive cells reveals for the first time that extramedullary PML-RAR-positive cell reservoirs such as the brain may persist and be involved in the leukemia relapse. We aimed at investigating the immune responses involved in the anti-leukemic effect of the combined immutherapy. To evaluate the...
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Receptors of NK cells
Hernychová, Lucie ; Novák, Petr (advisor) ; Pavlů, Barbora (referee)
NK cells play an important role as a part of the innate immune system, they however share many common features with cells of the adaptive immune system and they can modulate their functions as well. NK cells recognize non-self molecules (induced by viral infection or tumor transformation) through a wide variety of receptors on the cell surface. They also express receptors specific for their own structures, MHC glycoproteins. NK cells distinguish infected or transformed cells according to the quantity of these molecules on the target cell surface. Then NK cells allow apoptotic signals to cause cell death or the tolerance is established. They also promote inflammatory responses by the production of cytokines and chemokines. NK receptors can be grouped into activating, inhibitory, adhesion, cytokine, and chemokine receptors depending on their function. Based on interactions with appropriate ligands, NK cells exert cytotoxicity or they are inhibited. Moreover, their functions are influenced by the cytokine microenvironment. NK receptors can be also divided into C-type lectin and immunoglobulin superfamilies according to the structure. Ly49, NKG2/CD94 and NKR-P1 are receptor families that belong to the C-type lectin glycoproteins. These molecules have a type II transmembrane protein orientation and they...
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Host immune response in neuroinfections caused by parasitic helminths.
Macháček, Tomáš ; Panská, Lucie (advisor) ; Štáfková, Jitka (referee)
Among helmiths, there are some parasites which may dwell in the central nervous system of humans during their ontogenic development. Contrary to the former concept of so called immune privilege of this site, hosts develop the immune response against them. Both immune cells and resident immunocompetent cells (astrocytes, microglia) participate in these immune reactions. The host immune response consists of cellular as well as humoral immunity. This bachelor's thesis gives the general features of the immune response in the central nervous system and describes the immune response during particular helminthic neuroinfections: neuroschistosomiasis, neurocysticercosis and neurotoxocarosis.
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Molecular genetics of celiac disease
Němečková, Iva ; Daňková, Pavlína (advisor) ; Tučková, Ludmila (referee)
Celiac disease is an organ-specific autoimmune disease that arises as a consequence of hypersensitivity to the grain gluten in genetically susceptible individuals. Genetic predisposition are HLA-DQ2 and HLA-DQ8 genes, which are necessary but not sufficient for the emergence of celiac disease; it is involved in approximately 40% of the inheritance. In the course of the time, other genes that might contribute to the pathogenesis of celiac disease are being discovered. Among these so-called candidate genes, which are sought on the basis of known knowledge of molecular mechanisms of innate and adaptive immune responses, are for example: MIC, TNF, CTLA-4, CD28, ICOS, MYO9B, MMP, TLR and PTPN22. Immune response triggered by gluten peptide penetration into the lamina propria leads to mucosal damage. Different gluten peptides are involved in the pathology of celiac disease in different ways, some peptides trigger an adaptive immune response, while others, such as peptide p31- 43, triggers an innate immune response.
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IGFBP3 expressing rekombinant vaccinia virus used for tumor therapy
Musil, Jan ; Němečková, Šárka (advisor) ; Forstová, Jitka (referee)
IGFBP-3 expressing rekombinant vaccinia viruses used for tumor therapy Insulin-like growth factor-binding protein-3 (IGFBP-3) is a major regulator of endocrine effects of IGF and is capable to suppress the growth of variety of cancer. Several studies have shown that IGFBP-3 can induce the apoptosis of cancer cells via IGF-dependent and IGF-independent mechanisms. In our study, we have constructed recombinant vaccinia viruses (VACV) expressing IGFBP-3 under the control of the early H5 and synthetic early/late (E/L) promoter to investigate the potential effect on cancer growth in our cervical cancer model. We have shown that the expression of IGFBP-3 alone had no effect on tumor growth. On the other hand, the co-expression of IGFBP-3 enhanced the anti-cancer effect of immunization with the fusion protein SigE7LAMP, which gave rise to the anti-cancer immunity directed against HPV16 induced tumors. We have shown that the double-recombinant P13-SigE7LAMP-H5-IGFBP-3 can enhance the protective immune responses against MK16/ABC induced tumors. Furthermore, we have show that both double-recombinant viruses P13-SigE7LAMP-H5- IGFBP-3 and P13-SigE7LAMP-E/L-IGFBP-3 can increase the anti-cancer effect of SigE7LAMP expression in the therapy of TC-1 induced tumors. Key words: IGFBP-3, IGF, VACV, HPV16, E7 oncoprotein,...
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The role of humoral factors in the snail immune response against schistosomes
Košťáková, Monika ; Dvořák, Jiří (referee) ; Horák, Petr (advisor)
Digenetic trematodes such as Schistosoma mansoni use molluscs, mainly Gastropoda in their life cycle, as their intermediary hosts. e internal defense system (IDS) of snails is composed of immune cells called hemocytes, which are the main effectors and act jointly with soluble components. Humoral factors could in uence directly the parasite's larval stage, the activity of hemocytes and also may serve in recognition of the parasite. Lectins are considered to be the main component of humoral immunity. ey have a primary role in non-self recognition. Other protein group with lectin-like activity called FRePs was found in Biomphalaria glabrata. eir unique structure contains a brinogen and an immunoglobulin-like domain. Cytokine-like molecules may play very important role in defense as well. Many molecules are present in hemolymph and their levels change during infection. e response to parasitosis is therefore very complex and still awaits further clari cation.
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Tick-borne encephalitis - from pathogenesis to therapy
PALUS, Martin
The proposed thesis contributes to the knowledge about tick-borne encephalitis and its pathogenesis. The thesis describes pathogenesis and immunopathogenesis of tick-borne encephalitis, impact of host's genotype in clinical course determination, immune response of patients with acute tick-borne encephalitis, the mechanism of tick-borne encephalitis virus migration into central nervous system and virus interaction with cells of neurovascular unit as well as potential medical interventions.
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Efekt ADGF-A RNAi na imunitní odpověď \kur{Drosophily melanogaster} po infekci parazitickou vosou
ŠNEBERGEROVÁ, Pavla
In previous studies the extracellular adenosine has been proved as an important signaling molecule controlling dynamics of the immune response in Drosophila melanogaster. In mammals the adenosine signaling and control is more complex process. Since there is a certain genetic and evolutionary conservation between mammals and invertebrates, Drosophila represents simple system how to study such a complex regulation in vivo. Mutations of adenosine receptor in all cells as well as downregulation of ENT2 in immune cells caused decrease of resistance of the immune system against Drosophila parazitoid. The aim of the thesis is to determine whether RNAi targeted against mRNA of adenosine deaminase ADGF-A, which degrades e-Ado under natural conditions, would have any effect on immune response.
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