Transplantation is the best therapeutic solution for patients with chronic renal failure. Due to the great advances in immunosuppressive therapy in the last decades, graft and patient survival have improved significantly. On the other hand, immunosuppressive therapy has serious side effects - too strong immunosuppression may lead to infection or malignancies, conversely insufficient immunosuppression may lead to graft rejection. Due to the grave consequences of acute rejection, the main goal of cooperation of clinicians and transplant immunologists is to stratify patients into groups with low, moderate and high risk of rejection based on the evaluation of various immunologic risk factors. There are reports in the literature that the numbers (frequencies) of interferon gamma (IFNγ) producing cells before transplantation may be helpful to identify patients with high risk of acute cellular rejection and to predict long-term survival of the graft. In this retrospective study we determined the pre-transplant frequencies of activated donor specific T lymphocytes producing IFNγ after short stimulation (24 hrs) by ELISpot (Enzyme-linked immunosorbent spot assay). The results were correlated with the incidence of acute cellular (ACR) and antibody-mediated (AMR) rejection and with other risk factors. In our...

Current immunogenetic tests before organ transplantation include HLA typing and detection of HLA-specific antibodies. However, these tests do not provide information about the B cells participating in the humoral response against the transplanted organ. BAFF (B activating factor) plays an important role in the proliferation, maturation and differentiation of B cells. A soluble form of the cytokine arises after splicing the membrane form of BAFF. The soluble cytokine binds to three types of receptors - TACI, BCMA and BCMA. Some recent studies suggest that BAFF could serve as a marker or predictor of antibody-mediated (humoral) rejection in kidney transplant recipients. Our study consists of two parts. The first part is focused on the detection of soluble BAFF levels in patients after renal transplantation. The aim of our study was therefore to correlate levels of soluble BAFF cytokine in patients before and after transplantation with the clinical course and incidence of rejection after transplantation. The study included 92 kidney recipients. Humoral rejection was diagnosed on the basis of a positive finding of C4d deposits in peritubular capillaries (imunoflorescenční detection), and the presence of donor- specific antibodies. BAFF levels were determined using Xmap methodology by the Luminex method...

and Key words This diploma thesis is focused on assessing the potential benefit of HLA epitopes for the prediction of de novo antibody production at kidney transplant recipients. The topic and patient selection criteria were selected in accordance with the 18th International HLA and Immunogenetics workshop (IHIWS), which is taking place in May 2022 in the Netherlands, where our data will also be contributed. Our aims were to compare HLA antigens mismatches (counted as total number of mismatched alleles) defined on the high-resolution level by NGS sequencing, HLA eplets mismatches defined by HLA matchmaker, and amino acid mismatches defined by HLA EMMA in their capacity to predict de novo antibody production and compare these results to other works by different authors from this field. We have identified N= 28 patients who developed de novo antibodies and N= 19 who didn't develop de novo antibodies in 5 years follow up their transplant. These two cohorts were compared based on all three approaches and correlation between number of mismatches and number of patients with de novo antibodies were made using ROC curves. Superiority of eplet mismatches over HLA antigen mismatches (total number of mismatched alleles) defined on high resolution was not detected. The HLA epitopes identified by the HLA...

Memory T cells represent a specific subpopulation of cells formed during the first encounter with antigen. The main role of these cells is to elicit faster and more effective secondary response during reinfections. In transplant immunity, they may affect graft survival directly with donor-specific memory T cells or with cross-reactive virus-specific memory T cells. In this study, we focused on donor-specific and CMV-specific memory/effector T cells. We were interested in the effect of immunosuppressive therapy on the frequency of these cells in periphery. We found that the immunosuppression, prophylaxis and length of dialysis did not significantly affect the number of CMV-reactive cells 6 months after transplantation. We were also interested in the cross-reactivity between CMV and donor antigens, so-called heterologous immunity, which we verified by analyzing the TCR-β repertoire using next- generation sequencing (NGS) in CMV and donor-reactive T cells. Functional cross-reactive T cell clones (shared the same TCR-β sequence) were then found both in the peripheral blood of pre-transplant patients and in the post-transplant graft biopsy. We were also interested if long-term dialysis treatment affects immune memory. Dialysis therapy is often associated with the presence of poorly defined immune system...

86 Abstract - Transplantation Act: ethical and legal aspects. Theory and practice in the Czech Republic in comparison to the legal regulation in Germany Transplantations and related issues form an important part of medical law which is constantly developing. Currently it is a widely debated topic because there is lack of organs for donation, thus current supply can't satisfy all the demanding donees. It is a problem where the ethic, legal, medical and economic factors meet so there is no easy solution for majority of difficulties associated with transplantations. This diploma thesis focuses on contemporary legislation of organ and tissue donation and transplantations in Czech Republic which is mainly based on transplantation law and several legally binding international agreements. The aim of the thesis is a comprehensive evaluation of the current legislation and its comparison with the same legal branch in Germany. The work will also point out some of the shortcomings associated with the transplantation law and proposal of possible solution. Thanks to the comparison the thesis comes to a conclusion that the Czech transplantation law is relatively of high quality, but there are still imperfections which haven't been erased even by the approved amendatory act. However with relation to the German...

8 ABSTRACT Author: Bc. Lucie Moravcová Supervisor: MUDr. Vít Řeháček Charles University, Faculty of Pharmacy in Hradec Králové Title of master's thesis: Kidney transplantation: donor-recipient pairing in University Hospital Hradec Králové Background: The aim of this study was to determine HLA, blood group, age and sex match in donor-recipient pairing in kidney transplantation. HLA alleles of deceased donors were typed in Transfusion Department of the University Hospital Hradec Králové. Methods: Donor's HLA-A*, HLA-B* and HLA-DRB1* alleles were typed by PCR - SSP method. Complex data evaluating and processing was then performed. Results: 97 deceased donors were tested between 2013 and 2018. A total of 98 kidneys received from them were subsequently transplanted to 98 recipients in University Hospital Hradec Králové. 60,2 % of the donors were men, 63,3 % of the recipients were men. Most of the donors, as well as the recipients, were 51-70 years old (50,0 % and 59,2 %, respectively). The most common diagnoses in the group of deceased donors were associated with brain damage (66,3 %), the most common cause of renal failure in the group of recipients was chronic inflammatory kidney disease (41,8 %). All 98 transplantations (100,0 %) were AB0 compatible. 74 transplantations (75,5 %) were RhD compatible. 5...

Multiple myeloma is an agressive hemato-oncological disease the diagnosis and treatment possibilities of which have been developing for the last 15 years. The diagnostic methods include flow cytometry which uses antigen detection to distinguish between pathological and physiological plasmocytes. One of the monitored markers is marker CD45 which could be, according to our monitoring, of a negative prognostic value. 71 patients in our group were divided according to the intensity of CD45 antigen expression and the group with a lower expression showed a statistically higher risk of relapse within 12 months 62% vs. 25 % (p=0,0011). We did not find any connection with the influences of induction therapy or cytogenetics which are otherwise considered the most important prognostic markers. Multiple myeloma treatment involves a lot of combined protocols; and the repeated autologous transplantation is still considered to be the most efficient. We observed more positive results in planning the second autologous transplant early in patients who reached just partial remission after the 1st autologous transplant - significantly better TFS (treatment free survival) and overall survival (OS) were in the group of patients with early tandem transplantation: 18 months vs. 10 months (p=0.04) and the OS median was not reached...

The diagnosis of antibody-mediated rejection after liver transplantation is complicated, due to the fact that the clinical and pathological signs of this life- threatening complication are often overlapping with non-immunological symptoms, like biliary obstruction, ischemia, thrombosis and others. Furthermore, the transplanted liver is to a great extent resistant to this type of rejection. Like in the transplanted kidney and heart, the main pathological factors of graft injury are antibodies directed to the mismatched HLA antigens of the organ donor, i.e. donor- specific antibodies. Besides analysis of HLA specificity of antibodies, research lately has been directed to define whether these antibodies are complement-binding or not. Literature data on this are however up till now limited. Therefore, the aim of this diploma thesis was to study the clinical relevance of complement-binding antibodies against HLA antigens in patients after liver transplantation. Our preliminary results suggest that there might be a correlation between the presence of complement-binding antibodies and the development of antibody-mediated rejection. This finding may play a role for improvement of the prognosis of patients after liver transplantation. Key words HLA, antibodies, C4d, complement, liver transplantation

Gender differences in myocardial apoptosis of the patients after heart transplantation Background: Many functions of the cardiovascular apparatus are influenced by gender. The aim of our study was to find out the sensitivity to perioperative ischemia of the donor female and male myocardium; and determine how the organism affects the donor myocardium of the other sex after heart transplantation (detection of apoptosis), and whether the investigated biomarkers can predict primary graft dysfunction (PGD). Methods: The research was divided into three prospective studies. The Study 1 included 81 patients undergoing heart transplantation from September 2010 to January 2013. Patients were divided into two groups according to male allograft and female allograft. In order to prove myocardial necrosis the high-sensitive cardiac troponin T (hs-cTnT) method was used. Apoptosis was determined by immunohistochemical detection of caspase-3, Bcl-2, and by the TUNEL method. The Study 2 includeded 58 patients divided into four groups according to gender; both of the recipient and the donor. Apoptosis (caspase-3, Bcl-2, TUNEL) was analysed in these groups during the two-year follow-up. Into Study 3 64 patients were involved. We investigated the relationship in between these biomarkers and the development of PGD after...

Photoprotection in Patients after Kidney Transplantation Author: Zuzana Hajšelová Tutor: doc. PharmDr. Josef Malý, PhD. Consultant: Mgr. Barbora Vaňková Department of Social and Clinical Pharmacy, Faculty of Pharmacy in Hradci Králové, Charles University Introduction and aims: Patients after kidney transplantation (KT) are in a higher risk of developing skin cancer as a result of lifelong immunosuppressive (IS) therapy. Thorough photoprotection is therefore recommended. The aim of this study was to determine patients' awareness and level of photoprotection and to analyse selected risk factors for skin cancer development in patients after KT. Methods: The cross-sectional study was undertaken from 15th April to 31st December 2019 at the Haemodialysis Centre in the Teaching Hospital Hradec Králové. Patients included in this study were above 18 years old, who visited Haemodialysis Centre in the defined period. The data was collected from the patients' medical documentation (e.g. occurrence of skin cancer and skin cancer type, current IS) and from the written version of questionnaires (e.g. phototype, sun exposure, photoprotection). The data was analysed using descriptive statistics. Results: There were 410 KT patients in regular care of the Haemodialysis Centre, 361 fulfilled the questionnaire (88.0...