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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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Examination of genetic predispositions for celiac disease in a family with a history of diabetes mellitus 1. Type
KUČEROVÁ, Klára
Celiac disease is one of the autoimmune disorders that mainly affects the mucous membrane of the small intestine. The disease is characterized by intolerance to gliadin, which forms part of gluten. The intolerance leads to chronic inflammation of the mucous membrane of the small intestine, causing chronic diarrhea, adipose feces, vomiting, fatigue, and last but not least also weight loss. Type I. diabetes mellitus is an autoimmune metabolic disease demonstrated by hyperglycemia (increased level of blood glucose) which is a result of the insufficient effect of insulin. Type I. DM can occur at individuals as an independent disease or combined with other autoimmune diseases such as celiac disease. While genetic predisposition is a condition for the development of celiac disease, the development is influenced to a greater extent by external factors, for example environmental elements. Genetic predisposition is bound to alleles of the HLA system. This concerns in particular the HLA-DQ2 and HLA-DQ8 haplotypes. Due to various clinical symptoms, it is very difficult to diagnose the disease in some cases, and many patients are not diagnosed with the disease at all. The aim of the theoretical part of the bachelor thesis is to study and then summarize findings concerning celiac disease and type I. diabetes mellitus (DM I). To a great extent, the thesis addresses the topics of diabetes, celiac disease, and the HLA system. As for celiac disease, the thesis focuses on its characteristics, diagnostics, treatment, and inheritance. In the case of diabetes, the thesis deals with its individual types, treatment and also the genetic predispositions. The conclusion of the thesis describes the HLA system itself and shows the importance of the association between the type I. diabetes mellitus and celiac disease. The practical part of the thesis was conducted in the GENLABS genetics laboratory in České Budějovice. In this laboratory, genetic predispositions to celiac disease were examined by using the real-time PCR method. The examination is based on the HLA typing of the risk alleles by means of the commercially produced EliGene? Coeliac RT kit (DQ2, DQ8, DR4). The goal of the practical part is to master the basic laboratory methods such as the DNA isolation from the peripheral blood and buccal swab, to measure the DNA concentration, to adopt the real-time PCR method that serves to determine the HLA risk haplotype and to analyze gained results.
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Molecular genetic testing for celiac disease-associated HLA alleles
SALZMANOVÁ, Kateřina
Celiac disease is an autoimmune disease, which is characterised by intolerance of gliadin creating a part of gluten. Its intolerance leads to a chronic inflammatory response in the mucous membrane of small intestine, followed by malabsorption characterized by chronic diarrhoea, greasy stool and failure to thrive. Celiac disease is a multifactorial disease, caused by external environment factors and genetic factors. Genetic predisposition is given by HLA alleles HLA-DQ2 (DQA1*05/DQB1*02) or DQ8 (DQA1*0301/DQB1*0302) or HLA-DRB1*04. This disease has got a set of unspecified clinical symptoms, that is why most of the patients are not diagnosed at all. Estimated count of the ill people is up to 50 000, however real patients are only 15 % of them. The aim of the theoretical part of the bachelor thesis was to sum up the findings about the celiac disease and molecular genetic methods based on PCR. In the theoretical part about the celiac disease, I deal mainly with the description, types, diagnostics, pathogenesis and also with the diseases associated with celiac disease. Moreover, I dealt with the description and the nomenclature of HLA system and its association with the diseases. In the end of the theoretical part, I dealt with molecular genetic methods based on PCR (PCR-SSP and RT-PCR). I carried out the practical part of the thesis in a genetic laboratory at the Faculty of Health and Social Sciences, at the Faculty of Agriculture and at GENLABS, s.r.o. in Budweis. I dealt with the HLA typisation of alleles associated with the celiac disease. I did the examination by means of two methods - PCR-SSP method with the help of HISTO TYPE Celiac Disease kit and RT-PCR method with the help of EliGene? Coeliac RT kit (DQ2, DQ8, DRB4). The aim of the work was to embrace correct laboratory practice in a genetic laboratory. Then also set and optimize a commercially sold kit for an examination of HLA alleles associated with the celiac disease on the PCR-SSP principle. Examine the already examined patients by means of RT-PCR method. Compare both methods on the terms of accordance and difficulty.
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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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