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Methods of assesment and treatment of coeliac disease
Valkus, Martin ; Omelková, Jiřina (referee) ; Vránová, Dana (advisor)
This diploma thesis deals with methods of determining and treatment of coeliac disease. In the theoretical part of the work sums up informations about coeliac disease - history, prevalence, etiopathogenesis, immunopathogenesis, possibilities of diagnosis, discusses about gluten-free diet, mentions legislation of the Czech Republic and the European Union and compares expensiveness of gluten-free diet and normal diet. The experimental part of this thesis compares PCR, ELISA and indirect immunofluorescence methods for assesment of determination of genetic predisposition and antibodies in coeliac disease (greatest emphasis was placed on the comparison of antibodies against gliadin and deamidated gliadin antibodies in IgA and IgG).
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Imunomagnetic separation of lactic acid bacteria using magnetic microparticles functionalised by antibodies
Vaňásek, Jakub ; Španová, Alena (referee) ; Trachtová, Štěpánka (advisor)
Immunomagnetic separation is based on binding of antibody with antigen, where antibody is bound to magnetic particle. In this thesis there were used particles of magnetic pearl cellulose with antiLactobacillus and antiBifidobacterium antibodies. Immunomagnetic separation method was optimalized and verified for its efficiency and specifity with bacterial and yeast cells. This cells were identified by polymerase chain reaction. Efficiency of immunomagnetic separation was verified on probiotic meat product, where Lactobacillus cells were isolated. With DNA from isolated Lactobacillus cells the high resolution melting was performed. The results show presence of several bacterial strains of Lactobacillus species.
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Synthesis of quantum dots for proteins detection
Šibíková, Anna ; Fohlerová, Zdenka (referee) ; Pekárková, Jana (advisor)
This thesis is focused on synthesis of quantum dots (QDs) for protein detection. It comprises three parts. The first part summaries the theory of QDs, their synthesis, functionalization, interactions and applications in medicine. In the second part synthesis of CdTe/ZnS core/shell QDs modified by glutathione (GSH) is described, followed by the conjugation with biomolecules BSA and IgG. Several coupling agents such as EDC with NHS and CDI were used. In the last part, the final products were characterized by fluorescence spectroscopy and capillary electrophoresis. The results show the dependence of the fluorescence intensity of the QDs on pH range, concentration of BSA and IgG concentrations using different crosslinkers.
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Role of antibodies against HLA and non HLA antigens for organ transplantation.
Svobodová, Eva
Rejection is a significant complication after transplantation and one of the main reasons for loss of graft function. It is triggered by the response of the organ recipient's immune system based on the recognition of mismatched HLA (Human Leukocyte Antigens) and non-HLA antigens of the donor. All components of the immune system participate in this process, and according to the predominance of individual reactions, rejection can be divided into T cell-mediated rejection (TCMR) and antibody-mediated rejection (AMR). Rejection can develop immediately after transplantation in an acute form, or as a chronic form during several to tens of years after transplantation. The diagnosis of rejection is determined according to the clinical status, laboratory tests (including the detection of donor-specific antibodies, DSA) and histological findings in biopsies. The knowledge of validated gene expressions from the molecular microscope (MMDX) and other diagnostic tests has been recently applied. Individual phenotypes of rejection are evaluated and revised according to the international pathological classification. This work focuses on the analysis of immunological factors in relation to T-cell and antibody- mediated rejection after organ transplantation. The thesis deals with the determination of DSA in relation...
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Detection of soluble endogline in Lab-in-Syringe
Ilićová, Marie ; Svobodová, Zuzana (advisor) ; Chocholouš, Petr (referee)
The main target of the thesis was to develop an automated method for the isolation of endoglin from a sample using a magnetic immunosorbent in Lab-In-Syringe with subsequent immunodetection by affiblot. The theoretical part deals with endoglin, in its membrane and soluble form, especially with their connection to endothelial dysfunction. The principles of anti-endoglin therapy using the monoclonal antibody carotuximab or TRC105 are also briefly described here. Furthermore, the theoretical part summarizes the basics of the automation in Lab-In-Syringe and of the affiblot technique, their principles, and their applications. The last chapter deals with the biofunctionalization of magnetic particles by antibodies in order to prepare a magnetic immunosorbent which is used for the isolation of proteins from a complex sample. The experimental part describes methods for testing the affiblot prototypes. The main goal was to develop a method for the preparation of anti-endoglin magnetic immunosorbent and to increase the capacity of the method by upscaling it from a 1 ml to a 5 ml syringe. Furthermore, methods for the isolation of the soluble form of human endoglin from the culture medium containing the drug TRC105 were developed for 1 ml and 5 ml syringes. Various experiments were carried out to optimize the...
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Adaptive immune response against BK polyomavirus infection
Rezlerová, Adéla ; Saláková, Martina (advisor) ; Roubalová, Kateřina (referee)
BK polyomavirus is a small non-enveloped virus that is found in a large proportion of the human population. BKPyV infection commonly occurs in early childhood. The virus establishes persistent infection in renal tubular cells and uroepithelial cells. In immunosuppressed individuals, especially after renal or haematopoietic stem cell transplantation, BKPyV reactivation can lead to serious complications such as BKPyV-associated nephropathy (BKVAN), urethral stenosis or haemorrhagic cystitis. Adaptive immunity plays a crucial role in controlling the replication and progression of BKPyV infection. The T cell response is particularly important, with the production of CD4+ and CD8+ T-lymphocytes. B-lymphocytes, which produce virus neutralising antibodies, are also important. Currently, there are no effective antiviral agents against BKPyV infection and reducing immunosuppression remains the main strategy to suppress reactivation. Exploration of immune- based therapies offers promising possibilities for effective treatment of complications associated with polyomavirus infection. Key words: BK polyomavirus, T cell response, antibodies, BK polyomavirus-associated nephropathy, hemorrhagic cystitis
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Role of antibodies against HLA and non HLA antigens for organ transplantation.
Svobodová, Eva ; Slavčev, Antonij (advisor) ; Mrázek, František (referee) ; Černá, Marie (referee)
Rejection is a significant complication after transplantation and one of the main reasons for loss of graft function. It is triggered by the response of the organ recipient's immune system based on the recognition of mismatched HLA (Human Leukocyte Antigens) and non-HLA antigens of the donor. All components of the immune system participate in this process, and according to the predominance of individual reactions, rejection can be divided into T cell-mediated rejection (TCMR) and antibody-mediated rejection (AMR). Rejection can develop immediately after transplantation in an acute form, or as a chronic form during several to tens of years after transplantation. The diagnosis of rejection is determined according to the clinical status, laboratory tests (including the detection of donor-specific antibodies, DSA) and histological findings in biopsies. The knowledge of validated gene expressions from the molecular microscope (MMDX) and other diagnostic tests has been recently applied. Individual phenotypes of rejection are evaluated and revised according to the international pathological classification. This work focuses on the analysis of immunological factors in relation to T-cell and antibody- mediated rejection after organ transplantation. The thesis deals with the determination of DSA in relation...
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Chicken antibodies against the pathogenic yeast Candida albicans
Konečná, Lucia ; Heidingsfeld, Olga (advisor) ; Kubíčková, Božena (referee)
5 Abstract Candida albicans belongs to the most widespread pathogenic yeasts. As an opportunistic pathogen, it mainly colonises the mucous membranes of the host and causes infection, especially in individuals with a compromised immune system. The most common infection caused by C. albicans is vaginal mycosis, which at least 3 out of 4 women will experience during their lifetime. Antifungal drugs are used to treat this condition, but the yeasts are becoming less susceptible. For this reason, there is an effort to develop new treatment options for (not only) infections caused by C. albicans. Although vaginal mycosis is not a severe condition in most cases, it is a widespread infection that occurs repeatedly in many women, sometimes several times a year. Since vaginal mycosis reduces the quality of life in many women, we decided to focus on this disease in this paper. This project used hen antibodies, which have a number of advantages over mammalian antibodies. In oviparous animals, antibodies are not only found in the blood serum, as seen in mammals but are also secreted into their eggs. This makes the isolation of hen antibodies non-invasive and ethical. Moreover, the methods of isolating antibodies from eggs are also simpler than isolation from blood serum. Ultimately, hen antibodies are also more...
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Pre-transfusion laboratory examination of patients with AIHA and multiple myeloma
INGRIŠOVÁ, Karolína
This bachelor thesis follow up laboratory pre-transfusion tests in the special imunohematology lab for patients with AIHA diagnosis and multiple myeloms. For said patients, compatibility of the erythrocyte tranfusion must be maintained using special procedures. For AIHA patients, that means removal of patient's auto-antibodies by saturating the patient's plasma sample. As for patients with multiple myeloma, treatment of erythrocytes (either diagnostic or donor) with DTT is needed to remove false positivity caused by other treatments (such as administration of Daratumumab). Theoretical part consists of explanation of the basic immunohematological terms followed by thorough description of tests needed for patients with mentioned problematic diagnosis. Practical parts describes all laboratory procedures of each individual pre-transfusion tests including special ones for patients with AIHA and MM. This part also contains evaulation and analysis of all patients with AIHA and MM examined at the transfusion department of the University Hospital in Pilsen during the years 2021 and 2022. At the end of this part, all aquired data are evaluated using charts and graphs. End of this thesis contains summary and importance of tests performed for patients with the diagnosis of AIHA and MM, including answers for previously set research questions.
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Biomarkers of neurodegeneration in serum and cerebrospinal fluid in patients with selected neurological diseases
Nosková, Libuše ; Fialová, Lenka (advisor) ; Tlaskalová - Hogenová, Helena (referee) ; Švarcová, Jana (referee)
Neurofilaments are the key structural component of the cytoskeleton of neurons, where they are essential for many functions. They consist of 3 subunits: light chain (NFL); medium chain (NfM) and heavy chain (NfH). Except neurofilament proteins there is also α-internexin in the central nervous system (CNS) or peripherin in the peripheral NS. Due to various pathophysiological processes, neurofilament proteins are released into the extracellular space, where they can interact with the components of the immune system. While the involvement of the immune system in the pathogenesis of neurodegenerative diseases is obvious, less knowledge about the antibody response to the neurofilament proteins is available. It is eligible to expand our knowledge in this area. Determination of free antibodies against neurofilaments together with their immune complexes with corresponding antigen provides us more detailed insight into the antibody immune response against neurofilaments. We have optimized the ELISA methods to determine free antibodies against light and heavy chain of neurofilaments together with their corresponding immunocomplexes in both serum and cerebrospinal fluid. Implementation of these methods is precondition for analysis of those parameters in serum and cerebrospinal fluid of patients with...
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