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Markers of transplantation tolerance in kidney transplantation
Krepsová, Eva ; Viklický, Ondřej (advisor) ; Krejčí, Karel (referee) ; Živný, Jan (referee)
Long-term renal graft acceptance still requires long-term immunosuppressive therapy, which is accompanied by many adverse effects. Contrarily insufficient immunosuppression could lead to graft rejection and its failure. Therefore, research continues for biomarkers that reflect a patient's immunological status and thus allowing for individualized immunosuppressive therapy. In our study we showed lower incidence of acute rejection in kidney transplant recipients treated with rabbit anti-thymocyte globulin (rATG) or basiliximab induction within the first three months after transplantation. The rATG induction caused profound decrease of recipient's peripheral blood T and NK cells, as well as transcripts that are exclusively expressed by these cell types together with expansion of regulatory T cells (Tregs) among CD4+ T cells. In rATG group the increase of two transcripts associated with rejection (MAN1A1 and TLR5) was also observed in early post-transplant period. After the basiliximab induction we transiently detected CD4+CD25low/-FoxP3+ cell population along with disappearance of CD4+CD25+FoxP3+ Tregs. Basiliximab induction resulted in a transient increase in CD4+FoxP3+ Tregs, accompanied by the highest peripheral expression levels of markers associated with operational tolerance (FOXP3 and TCAIM)....
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Prediction of graft function development and rejection of transplanted kidney
Wohlfahrtová, Mariana ; Viklický, Ondřej (advisor) ; Zadražil, Josef (referee) ; Reischig, Tomáš (referee)
Improving the short-term results of kidney transplantation did not result in improving the long-term function and survival of kidney allograft. Organ shortage and increasing number of marginal donors remains the key problem in transplant today. The quality of donor organ is critical for graft function development and survival. The aim is to improve understanding to ischemia/reperfusion injury and its consequences, predict delayed graft function and rejection, improve organ allocation strategy and identify patients suitable for safe drug minimization or complete withdrawal of immunosuppressive therapy. Analysis of donor kidneys identified poor tubular cell quality and low survival factor, Netrin-1 expression levels, to be associated with delayed graft function. We confirmed that reperfusion phase of ischemia/reperfusion injury leads to minimal morphological but significant molecular abnormalities. Dissociation observed in histology and molecular pathology finding calls for an integrated approach in donor quality organ evaluation and allocation for transplantation. Significant heterogeneity within donors with expanded criteria was shown and subgroup of organs at low risk of delayed graft function was identified. We suggested donor biopsies to be performed as a routine praxis in all kidneys...
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Selected candidate genes variants with respect to the renal transplantation outcomes
Bandúr, Štěpán ; Viklický, Ondřej (advisor) ; Zadražil, Josef (referee) ; Reischig, Tomáš (referee)
6 Abstract: Renal transplantation outcomes might be influenced by genetic determinants, genes encoding structural and regulatory proteins involved in respective pathophysiological and immunological pathways. CYP3A4/CYP3A5 and ABCB1/MDR1 genes encoding cytochrome P450IIIA and P-glycoprotein, proteins determining bioavailability of immunosuppressants- calcineurin inhibitors and mTOR inhibitors, play important role. Polymorphisms in these genes have been suggested to influence immunosuppressants pharmacokinetics and renal transplant outcomes. The association of CYP3A4 -288A>G, CYP3A5 +6986G>A, ABCB1/MDR1 +1236C>T, +2677G>T>A and +3435C>T polymorphisms with clinical and laboratory endpoints was assessed by using haplotype analysis approach in 1016 patients. Except of HLA-DR mismatch, delayed graft function and kidney graft donor age, [ABCB1/MDR1+1236C;+2677G;+3435T] haplotype was independent predictor of acute rejection, but did not influence kidney graft survival. Homozygotes for the [CYP3A4- 288A;CYP3A5+6986G] haplotype had higher bioavailability of tacrolimus at week one. ABCB1/MDR1 haplotypes did not influence pharmacokinetics, but modified the risk of acute rejection, suggesting that allelic arrangement of the genetic loci was a stronger regulator of P- glycoprotein activity than single polymorphisms....
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Změny exprese signálních proteinů a genů v souvislosti s diabetickou nefropatií
Demová, Hana ; Černá, Marie (advisor) ; Viklický, Ondřej (referee) ; Kalousová, Marta (referee)
Hypothesis: I have investigated specific molecular and cell mechanisms that might be involved in the ethiopatogenesis of diabetic nephropathy. Their involvement might also be demonstrated in response to treatment of diabetic nephropathy. Aims: I have studied changes of signal transduction proteins (CAV-1, VEGF, RhoA) in renal cortical cells in the rat model of renal hypertension and in the rat model of type 1 diabetes. I also have performed analyses of polymorphisms in the genes involved in cell signaling with respect to their function in human study. Methods : The renal cortical expression of molecular targets CAV-1, VEGF, and RhoA have been assessed in addition to measurements of renal functional parameters in L-NAME-treated rats with or without a concomitant administration of atorvastatin (ATO) and have been compared to untreated control animals.
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Molecular Markers with Impact on Kidney Graft Survival and Glomerulopathies Progression
Brabcová, Irena ; Viklický, Ondřej (advisor) ; Jirsa, Milan (referee) ; Rychlík, Ivan (referee)
The progression of chronic glomerulopathy and graft rejection is affected by a number of proinflammatory cytokines, whose role in the pathogenesis of damage is poorly understood. The aim of this dissertation was to identify reliable risk markers of renal dysfunction progression and thereby contribute to a more effective patient treatment. Human native kidney biopsies with histologically confirmed diagnosis of glomerulopathy or kidney graft biopsies were analysed. Intrarenal gene expressions were measured by RT-qPCR. Single nucleotide polymorphisms were detected by methods based on PCR-RFLP. Immunohistochemical staining was used to identify and quantify the mononuclear cell infiltration. Gene expression of TGF-β1, HGF, BMP7, MCP-1, RANTES and mononuclear cell infiltration were associated with poor renal function and proteinuria at the time of IgA nephropathy diagnosis. Progression of IgA nephropathy during the 2-year follow-up was shown to be dependent on the degree of chronic vasculopathy and TGF-1 expression in the kidney. Patients with graft dysfunction and enhanced intrarenal expression of TGF-1, MCP-1 had significantly shorter graft survival. Higher mRNA expression of IL-10, TGF- 1, IL-6, MCP-1, RANTES and TNF- was observed in patients with graft dysfunction presented at the time of biopsy....
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Liver and kidney dysfunction in critically ill patients. Support options and compensation functions
Kroužecký, Aleš ; Matějovič, Martin (advisor) ; Chytra, Ivan (referee) ; Viklický, Ondřej (referee) ; Dostál, Pavel (referee)
22 23 7. Summary The liver and kidney are an important organs involved in a number of biosynthetic, biotransformative, detoxifying, endocrine and immune processes and therefore it is understandable that its dysfunction is associated with adverse outcome of critically ill patients. Hepatic and renal dysfunctions in critical illness are relatively common and occur usually as a component of multiple organ dysfunction syndrome (MODS). Several mechanisms have been proposed to explain the development of MODS. Among these, 1) the hypoxic component resulted from an inadequate oxygen supply to tissues, and 2) cytotoxic effects of various mediators are believed the key elements in the pathophysiology of MODS. From these complex causes of injury are probably the most therapeutically attainable the hemodynamic disturbances and maintenance of adequate organ perfusion pressure using vasopressors is one of the cornerstones of treatment of critically ill patients. Blood flow through organs is autoregulated over a wide range of mean arterial pressure (MAP). There is an agreement that under physiological conditions minimum value of MAP necessary to ensure this autoregulation is about 60- 65 mm Hg. Therefore MAP > 65 mmHg has been recommended as a goal for the vasopressor therapy in sepsis. In critical condition, however,...
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Cytomegalovirus infection after kidney transplantation
Reischig, Tomáš ; Třeška, Vladislav (advisor) ; Viklický, Ondřej (referee) ; Zadražil, Josef (referee) ; Pazdiora, Petr (referee)
1 SUMMARY Cytomegalovirus (CMV) disease is a common infectious complication in patients after solid organ transplantation. The last decade witnessed major advances in CMV disease prevention. Use of universal prophylaxis or preemptive therapy resulted in a decrease in the incidence of CMV disease from 20-60% to 5-20%. However, the efficacy of preventive approaches in terms of indirect effects of CMV occurrence is problematic. Association with allograft rejection belongs to well documented and clinically extremely important indirect effects of CMV with a prolonged adverse impact on graft survival. Potential mechanisms include overexpression of major histocompatibility complex molecules, growth factors and cytokines, and upregulation of adhesion molecules. A number of questions remain to be answered in evaluating CMV as a risk factor for acute rejection. While CMV disease is associated with an increased incidence of acute rejection, data regarding the role of asymptomatic CMV viremia are controversial. In our research we evaluated the role of CMV in pathogenesis of allograft rejection in the era of modern immunosuppression and CMV prophylaxis as well as optimal preventive strategy to minimize impact of CMV. In the first trial, renal transplant (RTx) recipients were followed prospectively for 12 months to...
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Molecular Basis of Hereditary Hyperuricaemic Nephropathies
Vyleťal, Petr ; Kmoch, Stanislav (advisor) ; Viklický, Ondřej (referee) ; Jirsa, Milan (referee)
Familial juvenile hyperuricemic nephropathy (FJHN) and medullary cystic kidney disease type 1 (MCKD1) and type 2 (MCKD2) are autosomal dominant tubulointerstitial nephropathies characterized by combinations of hyperuricaemia, gouty arthritis, progressive renal insufficiency, and in some but not all families, medullary cysts. The phenotypic expression of these diseases is inconsistent, overlaps and indicates broader genetic and allelic heterogeneity. Their pathophysiology was mainly unknown. Previous studies localized FJHN/MCKD genes to chromosomes 16p11 and 1q21. This thesis was primarily aimed at identification of molecular bases and mechanisms underlying FJHN/MCKD. To follow this aim, we focused on collection and characterization of FJHN/MCKD patients and families, identification of disease causing genes in affected families, characterisation of identified proteins and their mutated forms and the isolation and characterisation of interacting partners of newly identified proteins. We employed and established numerous molecular genetic, molecular biological and biochemical methods. We gathered one of the largest sets of families with FJHN/MCKD in the world. In about 26% of families we identified UMOD (uromodulin encoding) gene mutations and characterised by various approaches 6 uromodulin mutant proteins....
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