Preparation of fusion domains of human immunoreceptors for their utilization in immunotherapy

The functions of the immune system include immunosurveillance of transformed cells, i.e., the ability to eliminate these cells before they become harmful to the organism. If the transformed cells succeed to escape the immune system surveillance, an oncological disease develops. The tumour immunotherapy aims to stimulate the immune system mechanisms to fight against the tumour. Lately, there's an interest in using NK cells in the immunotherapy of tumours. These cells appertain to the innate immune system and participate in immunosurveillance. When an NK cell encounters a target cell, its activation depends on the integration of signals from the surface activating and inhibiting receptors which bind ligands on the surface of the target cell. Upon activation, NK cell exhibits a cytotoxic response against the target cell. The use of NK cells in immunotherapy includes, among others, the testing of bispecific fusion proteins which can bind a tumour surface antigen by one part and NK cell activating receptor by the other part. Thus, these fusion proteins mediate a contact between both cells and trigger the cytotoxic response. This work presents a preparation of bispecific fusion proteins which consist of an activating ligand MICA (for the receptor NKG2D) or B7H6 (for the receptor NKp30), and a nanobody...Jednou z funkcí imunitního systému je imunitní dohled nad nádorově transformovanými buňkami, tedy schopnost takové buňky odstranit dříve, než se mohou stát škodlivými pro organismus. V případě, že se transformovaným buňkám podaří imunitnímu dohledu uniknout, dochází k rozvoji onkologického onemocnění. Nádorová imunoterapie spočívá v cílené stimulaci imunitního systému v boji proti nádoru. V poslední době se projevuje zájem o využití NK buněk v imunoterapii nádorů. Tyto buňky, patřící k nespecifické imunitě, se podílejí na imunitním dohledu. Při setkání s cílovou buňkou rozhoduje o aktivaci NK buňky integrace signálu z jejích povrchových aktivačních a inhibičních receptorů, které vážou ligandy na povrchu cílové buňky. V případě, že dojde k aktivaci NK buňky, spouští proti cílové buňce cytotoxickou odpověď. Využití NK buněk v imunoterapii zahrnuje mimo jiné testování bispecifických fúzních proteinů, které mohou jednou částí vázat povrchový nádorový antigen a druhou částí NK buněčný aktivační receptor, čímž zprostředkují kontakt obou buněk a vyvolají cytotoxickou odpověď. Tato práce představuje přípravu bispecifických fúzních proteinů sestávajících z aktivačního ligandu MICA (pro receptor NKG2D), nebo B7H6 (pro receptor NKp30) a nanoprotilátky cílící jeden ze tří nádorových antigenů HER2, CD20 a FGFR....Department of BiochemistryKatedra biochemieFaculty of SciencePřírodovědecká fakult

Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse.

Signaling by the human C-type lectin-like receptor, natural killer (NK) cell inhibitory receptor NKR-P1, has a critical role in many immune-related diseases and cancer. C-type lectin-like receptors have weak affinities to their ligands; therefore, setting up a comprehensive model of NKR-P1-LLT1 interactions that considers the natural state of the receptor on the cell surface is necessary to understand its functions. Here we report the crystal structures of the NKR-P1 and NKR-P1:LLT1 complexes, which provides evidence that NKR-P1 forms homodimers in an unexpected arrangement to enable LLT1 binding in two modes, bridging two LLT1 molecules. These interaction clusters are suggestive of an inhibitory immune synapse. By observing the formation of these clusters in solution using SEC-SAXS analysis, by dSTORM super-resolution microscopy on the cell surface, and by following their role in receptor signaling with freshly isolated NK cells, we show that only the ligation of both LLT1 binding interfaces leads to effective NKR-P1 inhibitory signaling. In summary, our findings collectively support a model of NKR-P1:LLT1 clustering, which allows the interacting proteins to overcome weak ligand-receptor affinity and to trigger signal transduction upon cellular contact in the immune synapse

Problematika Mezinárodního managementu a Mezinárodní pracovní mobility

Cílem této práce je zamyšlení nad úspěšností expatriatů v jejich adaptačních procesech. Nejdříve práce nabízí teoretický rámec obsahující hlavně vybrané teorie mezinárodního managementu a mezinárodní pracovní mobility a pak přichází s vlastním výzkumem provedeným na 6 reálných expatriatech, kteří v současné době pracují v zahraničí. Práce zodpovídá otázky týkající se úspěšnosti adaptace; hlavně si pokládá otázku, jaké že jsou důležité osobní a profesní předpoklady k úspěšné adaptaci expatriata. Na základě detailní studie individuálních zkušeností této šestice, hlavní aspekty jsou diskutovány a později navrženy.The goal of this thesis is to elaborate on the aspects of expatriate's successfulness in adaptation process. Firstly presenting theoretical background consisting of theories of international management and international mobility and after that introducing a research with six expatriates currently being on an assignment. It answers questions regarding successfulness of adaptation; mainly what personal and professional traits are valuable when being an expatriate. Based on the detailed study of their personal experiences, the key factors are discussed and suggested

Financial Analysis of an Enterprise

Zpracování finanční analýzy společnosti ZVVZ a.s. (výpočet ukazatelů, časové srovnání a srovnání výsledků v rámci odvětví, ve kterém podnik působí).Processing of financial analysis ZVVZ a.s. (Calculation of parameters, temporal comparison and comparison of results across the sector in which the company operates

Benefit analysis of employee management and development by competencies

The paper presents the benefits of employee management and development by competencies. The work has a theoretical and empirical character and it is dividend into six parts. The first two parts focus on the theories of an organizational strategy and the concepts of the competency approach in HR management and development in general. The practical application of the organizational strategy and competency approach in HR processes of an international company Cemex is critically observed in the third and fourth part. The analysis conducted on the basis of a "focus groups" among managers of Cemex constitutes the fifth part of this paper. The purpose of this analysis is to objectify the benefits of competency approach of the management and development in this organization. The sixth part summarizes the conclusions of the paper. The paper discusses the benefits of employee management and development by competencies primarily with focus on managers. A partial aim of the text is an extension of analytical perspectives on the competency approach in theory and practice of an organizations. The main objective of the thesis, however, is the identification and analysis of the benefits resulting from the use of competency models and approaches in the assessment and development system of managers and shaping of..

Preparation of fusion domains of human immunoreceptors for their utilization in immunotherapy

The functions of the immune system include immunosurveillance of transformed cells, i.e., the ability to eliminate these cells before they become harmful to the organism. If the transformed cells succeed to escape the immune system surveillance, an oncological disease develops. The tumour immunotherapy aims to stimulate the immune system mechanisms to fight against the tumour. Lately, there's an interest in using NK cells in the immunotherapy of tumours. These cells appertain to the innate immune system and participate in immunosurveillance. When an NK cell encounters a target cell, its activation depends on the integration of signals from the surface activating and inhibiting receptors which bind ligands on the surface of the target cell. Upon activation, NK cell exhibits a cytotoxic response against the target cell. The use of NK cells in immunotherapy includes, among others, the testing of bispecific fusion proteins which can bind a tumour surface antigen by one part and NK cell activating receptor by the other part. Thus, these fusion proteins mediate a contact between both cells and trigger the cytotoxic response. This work presents a preparation of bispecific fusion proteins which consist of an activating ligand MICA (for the receptor NKG2D) or B7H6 (for the receptor NKp30), and a nanobody..

The consequences of specific learning disabilities for educational way and adulthood

Diplomová práce se zabývá základními východisky, deskripcí projevů specifických poruch učení a také reedukací. Jejím hlavním smyslem však je na pozadí celé školní dráhy dítěte sledovat jedno z klíčových období vývoje, a to jak specifické poruchy učení ovlivňují rozhodování o volbě povolání a jaké důsledky mají pro uplatnění v životě. Diplomová práce má charakter teoreticko-empirický, její součástí jsou tři kazuistiky dospělých lidí se specifickou poruchou učení

Requirements of management on sales reporting

The thesis maps the practices of contemporary sales reporting, analyses its weaknesses and looks for new and original solutions of maximal efficiency of reporting in today's companies. It connects several fields from management accounting, human resources and marketing to graphics and typesetting to get the maximal outcome and efficiency which is always needed in creating sales reports

Financial Analysis of an Enterprise

Processing of financial analysis ZVVZ a.s. (Calculation of parameters, temporal comparison and comparison of results across the sector in which the company operates