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Carbohydrate dimers in tumor immunotherapy
Krupová, Monika ; Bezouška, Karel (advisor) ; Ryšlavá, Helena (referee)
Carbohydrate dimers in tumor immunotherapy Monika Krupová (Department of Biochemistry, Faculty of Science, Charles University in Prague) One possible approach to tumor immunotherapy is an activation of killer lymphocytes through specific ligands for their surface receptors. CD69 is a molecule greatly widespread among various cells of haematopoietic origin. Since the physiological ligand for this receptor is still unknown, ligand mimetics are used for modulation of its activity. The mimetics tested in this work are based on monomeric or oligomeric carbohydrated attached through two different chemical groups to the central linker of varying length, giving rise to thiourea and triazole series. The ability to precipitate soluble NKR-P1 and CD69 receptors was evaluated in precipitation assays and the optimal length of the linker for NKR-P1 receptor was found to be decyl. On the other hand, cross-linking of CD69 is not so dependent on the length of the linker. The aim of this work was to describe in vitro effect of the tested compounds on cellular signalization, natural killing of leukemic cell lines and activation-induced apoptosis. Compounds of triazole series containing two disaccharides (GalNAc β1→4 GlcNAc) linked by a linker were found to have the strongest effect on the production of...
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Preparation of expression vector for production of receptors CD69 and S1P1
Leontovyč, Adrian ; Vaněk, Ondřej (advisor) ; Obšil, Tomáš (referee)
T- and B- lymphocytes play a key role in immunity, because they provide specific immunity in organism. Receptor S1P1 regulates lymphocyte egress from lymphoid organs to blood, from which the lymphocytes travel to the site of infection. It was discovered that transmembrane helix 4 of S1P1 receptor interacts with transmembrane part of CD69 receptor and this interaction is responsible for retention of lymphocytes in lymphoid organs. The aim of this study is to prepare expression plasmids for production of these two receptors and their further research.
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Interaction of Galectin-1 with human NK cell receptors
de Sousa Santos Abreu, Celeste ; Vaněk, Ondřej (advisor) ; Pavlíček, Jiří (referee)
Natural killer (NK) cells are a subpopulation of effector lymphocytes with cytotoxic activity and cytokine-producing functions considered as an integral part of the innate immune response. Functions of NK cells include tumour elimination, engagement and regulation of antiviral immune responses and regulation of immune cells by production and secretion of chemokines and cytokines. CD69 is a C-type lectin-like transmembrane receptor expressed in NK cells. CD69 is an activating receptor and acts also as a very early marker of lymphocyte activation. Putative protein ligands have been described for CD69 in the last years: Galectin-1, S1P1, S100A8/S100A9 and Myl9/12. Galectin-1 is a prototypical lectin characterized by the presence of a common lectin structural fold and a carbohydrate recognition domain involved in carbohydrate binding. Galectin-1 was identified as a binding partner for CD69 based on biological and functional studies, but structural details about the complex are still missing. This thesis describes the successful establishment of an expression protocol for a tag-less cysteine-less mutant of galectin-1 and the study of the interaction between galectin-1 and NK cell receptors. The interaction was studied using microscale thermophoresis and confirmed as dependent on the presence of a...
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Preparation of expression vector for production of receptors CD69 and S1P1
Leontovyč, Adrian ; Vaněk, Ondřej (advisor) ; Obšil, Tomáš (referee)
T- and B- lymphocytes play a key role in immunity, because they provide specific immunity in organism. Receptor S1P1 regulates lymphocyte egress from lymphoid organs to blood, from which the lymphocytes travel to the site of infection. It was discovered that transmembrane helix 4 of S1P1 receptor interacts with transmembrane part of CD69 receptor and this interaction is responsible for retention of lymphocytes in lymphoid organs. The aim of this study is to prepare expression plasmids for production of these two receptors and their further research.
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Recombinant production of bovine NK cell receptors
Böerová, Nikola ; Kavan, Daniel (advisor) ; Šulc, Miroslav (referee)
NK cells, which are part of the innate immune system, are increasingly gaining attention, especially due to their cytotoxic ability to kill tumor cells of certain lines and certain viral, bacterial or parasitic infestation of the body. They lay a role in organ transplantation, the fight against HIV and other autoimmune diseases. NK cells have been studied since the 70th of the 20th century, but the structures and physiological ligands of their receptors remain only partially understood, as does the exact role of these cells in the organism. They communicate with others through their receptors, that recognize the lack of expression of MHC class I glycoproteins on the surface of target cells, thereby preventing cell recognition by cytotoxic lymphocytes. This diploma thesis deals with the research of receptors from cattle (Bos taurus), which is not a traditional laboratory animal and my task was to contribute to research of the structure of this group of proteins. I dealt with the recombinant production of some of the most important representatives of NK receptors CD69, NKRP1 and NKG2D in bacterial cells. The findings published in this thesis are a continuation of my bachelor thesis and together can be beneficial for further research into structural proteins and thus may help as in veterinary medicine...
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Carbohydrate dimers in tumor immunotherapy
Krupová, Monika ; Bezouška, Karel (advisor) ; Ryšlavá, Helena (referee)
Carbohydrate dimers in tumor immunotherapy Monika Krupová (Department of Biochemistry, Faculty of Science, Charles University in Prague) One possible approach to tumor immunotherapy is an activation of killer lymphocytes through specific ligands for their surface receptors. CD69 is a molecule greatly widespread among various cells of haematopoietic origin. Since the physiological ligand for this receptor is still unknown, ligand mimetics are used for modulation of its activity. The mimetics tested in this work are based on monomeric or oligomeric carbohydrated attached through two different chemical groups to the central linker of varying length, giving rise to thiourea and triazole series. The ability to precipitate soluble NKR-P1 and CD69 receptors was evaluated in precipitation assays and the optimal length of the linker for NKR-P1 receptor was found to be decyl. On the other hand, cross-linking of CD69 is not so dependent on the length of the linker. The aim of this work was to describe in vitro effect of the tested compounds on cellular signalization, natural killing of leukemic cell lines and activation-induced apoptosis. Compounds of triazole series containing two disaccharides (GalNAc β1→4 GlcNAc) linked by a linker were found to have the strongest effect on the production of...
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