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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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Analysis of serological cross-reactivity between antibodies against BK polyomavirus variants
Caisová, Helena ; Horníková, Lenka (advisor) ; Roubalová, Kateřina (referee)
BK polyomavirus (BKPyV), which asymptomatically infects about 80 % of the human population, can reactivate in immunosuppressed patients after kidney transplantation and cause nephropathy. In the European population, there are predominantly two BKPyV sub- types, BKPyV-I and BKPyV-IV, which behave as different serotypes. Recipients who receive a kidney from a donor positive for a different subtype are at a greater risk of graft rejection. So far, it is impossible to distinguish between these two subtypes using a serological ELISA test as serum antibodies against these two subtypes cross-react. This work aimed to iden- tify epitopes on the major antigenic protein VP1, which are responsible for the binding of cross-reacting and/or subtype-specific antibodies. The identification of such epitopes could further lead to improvement in pre-transplant diagnostics and better management of pati- ents after transplantation. Therefore, two types of antigens composed of the VP1 protein of the BKPyV-IV subtype with introduced mutations characteristic of BKPyV-I in the DE and EF loops of VP1 were prepared and tested using an antigen competition assay with a panel of human sera. The results showed that the DE loop region is an immunogenic epitope that binds specific antibodies for BKPyV subtype I. Conversely, a mutation in...
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Studium celogenomové variability lidského cytomegaloviru.
Dvořák, Jan ; Tachezy, Ruth (advisor) ; Roubalová, Kateřina (referee)
This work is part of a project focused on the study of the variability of human cytomegalovirus (HCMV) among clinical isolates with the aim to map the geographical distribution of HCMV genotypes, reveal the relationships between genotypes and the severity of HCMV-associated diseases, and identify regions in the HCMV genome with a potential for use as diagnostic and therapeutic targets. Attention was paid to the development of the methodology for the preparation of the material for next-generation sequencing (NGS) from HCMV clinical isolates and evaluation of the obtained sequencing data. Blood and urine samples collected from hematopoietic stem cell transplantat recipients and congenitally infected children were analyzed. Samples suitable for NGS were sequenced by the Illumina platform and sequences were created by de novo assembly followed by mapping assembly. Urine samples in comparison to blood samples had higher yield of material for NGS. Of the samples positive for HCMV DNA (7 of 50) after amplification in the cell cultures, only one sample had high purity of the viral DNA (98%) while six samples had purity of less than 7%. The sample containing 98% of the viral DNA was fully sequenced and the sequence was compared to the sequences of other clinical isolates from Belgium in 11 polymorphic...
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Psychosomatic linking of functional disorders of the upper gastrointestinal system and their significance for therapeutic rehabilitation
Roubalová, Kateřina ; Bitnar, Petr (advisor) ; Strapková, Zuzana (referee)
This thesis addreses the prohlems of medically unexplained physical symptoms of upper part of gastrointestinal tract from gastroenteorological, psychiatrical and physiotherapeutical perspective, and introduces psyclliatrical diagnostical unit "somatoform disorder". Next, it focuses on the interrelations between locomotor system, gastrointestinal system and psychical make-up in terms of the comprehensive approach to these disorders in physiotherapy. In the experimental part of this thesis psychological self-report questionnaire SCL-90 was used for screening of psychopathology of 19 outpatients with functional gastrointestinal disorder. From 15 retrieved questionnaires only 3 showed higher rates of general psychopathology. Further, patients underwent a physiotherapeutical examination to elicit eventual presence of so called visceral patterns and global disorders of locomotor system, which could affect their problems. These changes (both visceral patterns and global disorders of locomotor system) were found in the majority of cases. Powered by TCPDF (www.tcpdf.org)
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Construction of various types of vaccines based on the structural proteins of mouse polyomavirus and analysis of immune response after their administration to mice
Hrušková, Veronika ; Forstová, Jitka (advisor) ; Roubalová, Kateřina (referee) ; Reiniš, Milan (referee)
Conclusion Humoral and cellular immune responses developed in mice after intranasal delivery of model mouse polyomavirus derived VLPs carrying epitope of enhanced green fluorescence protein (EGFP) Model chimeric EGFP-VLPs were purified and used for immunization of mice. Immune response of immunized animals was examined. No specific antibodies against EGFP protein, but high titers of specific antibodies against major structural protein VP1 were developed in the sera of immunized animals. Splenocytes derived from immunized animals secreted IL-2 and IFN-γ after their antigen (EGFP or VP1) restimulation. Proliferation of CD4+, but not CD8+ T cells from immunized mice after the stimulation with both EGFP and VP1 was observed. No EGFP specific cytotoxic activity of splenocytes from immunized mice was detected. The presentation of EGFP-VLPs in the context ofMHC class II was blocked by inhibitors of endo-lysosomal as well as proteasomal compartments. Changes in the numbers of CD25+Foxp3+ subpopulation of CD4+ T cells were observed in the spleens if immunized mice. Chimeric VLPs derived from mouse polyomavirus carrying epitopes of human Bcr-Abl fusion protein (Bcr-Abl VLPs) Chimeric Bcr-Abl VLPs carrying 171 amino acids sequence of Bcr-Abl protein (containing Bcr-Abl breakpoint region) were prepared. Chimeric VLPs...
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Studium celogenomové variability lidského cytomegaloviru.
Dvořák, Jan ; Tachezy, Ruth (advisor) ; Roubalová, Kateřina (referee)
This work is part of a project focused on the study of the variability of human cytomegalovirus (HCMV) among clinical isolates with the aim to map the geographical distribution of HCMV genotypes, reveal the relationships between genotypes and the severity of HCMV-associated diseases, and identify regions in the HCMV genome with a potential for use as diagnostic and therapeutic targets. Attention was paid to the development of the methodology for the preparation of the material for next-generation sequencing (NGS) from HCMV clinical isolates and evaluation of the obtained sequencing data. Blood and urine samples collected from hematopoietic stem cell transplantat recipients and congenitally infected children were analyzed. Samples suitable for NGS were sequenced by the Illumina platform and sequences were created by de novo assembly followed by mapping assembly. Urine samples in comparison to blood samples had higher yield of material for NGS. Of the samples positive for HCMV DNA (7 of 50) after amplification in the cell cultures, only one sample had high purity of the viral DNA (98%) while six samples had purity of less than 7%. The sample containing 98% of the viral DNA was fully sequenced and the sequence was compared to the sequences of other clinical isolates from Belgium in 11 polymorphic...
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Construction of various types of vaccines based on the structural proteins of mouse polyomavirus and analysis of immune response after their administration to mice
Hrušková, Veronika ; Forstová, Jitka (advisor) ; Roubalová, Kateřina (referee) ; Reiniš, Milan (referee)
Conclusion Humoral and cellular immune responses developed in mice after intranasal delivery of model mouse polyomavirus derived VLPs carrying epitope of enhanced green fluorescence protein (EGFP) Model chimeric EGFP-VLPs were purified and used for immunization of mice. Immune response of immunized animals was examined. No specific antibodies against EGFP protein, but high titers of specific antibodies against major structural protein VP1 were developed in the sera of immunized animals. Splenocytes derived from immunized animals secreted IL-2 and IFN-γ after their antigen (EGFP or VP1) restimulation. Proliferation of CD4+, but not CD8+ T cells from immunized mice after the stimulation with both EGFP and VP1 was observed. No EGFP specific cytotoxic activity of splenocytes from immunized mice was detected. The presentation of EGFP-VLPs in the context ofMHC class II was blocked by inhibitors of endo-lysosomal as well as proteasomal compartments. Changes in the numbers of CD25+Foxp3+ subpopulation of CD4+ T cells were observed in the spleens if immunized mice. Chimeric VLPs derived from mouse polyomavirus carrying epitopes of human Bcr-Abl fusion protein (Bcr-Abl VLPs) Chimeric Bcr-Abl VLPs carrying 171 amino acids sequence of Bcr-Abl protein (containing Bcr-Abl breakpoint region) were prepared. Chimeric VLPs...
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Gene Therapy of CML: Experimental Vaccines against Bcr-abl-transformed Cells
Lučanský, Vincent ; Vonka, Vladimír (advisor) ; Roubalová, Kateřina (referee) ; Reiniš, Milan (referee)
Chronic myeloid leukemia is malignant disease characterized by myeloproliferative clonal expansion of hematopoietic stem cell. It is causally associated with the formation of the so called Philadelphia chromosome and production of its specific product, the chimeric BCR-ABL protein. The amino acid sequence of the fusion region is unique, implying that the BCR-ABL protein carries tumor specific antigen. Currently imatinib mesylate dominates the treatment of CML. It is well tolerated and when compared to the other drugs used, it prolongs the life expectancy significantly. Unfortunately, it is not capable to cure the disease. The only potentially curative approach nowadays is the bone marrow transplantation; however, it is connected with a relatively high morbidity and mortality. Moreover, it is available only to a minority of the patients. Under these circumstances the need for the development of a relatively safe and generally available treatment is understandable. Immunotherapy could be such a treatment. Several experimental vaccines based on BCR-ABL sequence were developed and tested in mice in our institute. The DNA vaccines used were carrying sequences coding for the whole BCR-ABL protein, or for 25 amino acids long junction region (these DNA sequences were fused with adjuvant genes such as...
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Somatoform disorder and its influencing through physiotherapy
Roubalová, Kateřina ; Ocmanová, Renata (advisor) ; Hejlová, Věra (referee)
Subject of this thesis is to inform about the somatoform disorder and outline its position in physioterapy. On the basis of findings about this phenomenon, the possibilities of physioterapy are introduced, as well as some means which could be used to positively influence the somatoform disorder within the frame of complex approach along with the principles of treatment of patients with somatoform symptoms. Powered by TCPDF (www.tcpdf.org)
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Psychosomatic linking of functional disorders of the upper gastrointestinal system and their significance for therapeutic rehabilitation
Roubalová, Kateřina ; Bitnar, Petr (advisor) ; Strapková, Zuzana (referee)
This thesis addreses the prohlems of medically unexplained physical symptoms of upper part of gastrointestinal tract from gastroenteorological, psychiatrical and physiotherapeutical perspective, and introduces psyclliatrical diagnostical unit "somatoform disorder". Next, it focuses on the interrelations between locomotor system, gastrointestinal system and psychical make-up in terms of the comprehensive approach to these disorders in physiotherapy. In the experimental part of this thesis psychological self-report questionnaire SCL-90 was used for screening of psychopathology of 19 outpatients with functional gastrointestinal disorder. From 15 retrieved questionnaires only 3 showed higher rates of general psychopathology. Further, patients underwent a physiotherapeutical examination to elicit eventual presence of so called visceral patterns and global disorders of locomotor system, which could affect their problems. These changes (both visceral patterns and global disorders of locomotor system) were found in the majority of cases. Powered by TCPDF (www.tcpdf.org)
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