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Development of a procedure for the analysis of the T-cell response against SARS-CoV-2 virus
Šmahel, Michal ; Pavliš, Oto ; Beran, Ondřej ; Bohoněk, Miloš ; Tachezy, Ruth ; Holub, Michal ; Poláková, Ingrid ; Saláková, Martina ; Schreiberová, Lucie
In 2020, the SARS-CoV-2 coronavirus caused a pandemic of covid-19 disease. To detect the cellular response induced by infection with this virus, a highly sensitive ELISPOT assay has been developed that detects specific interferon gamma-producing T lymphocytes after stimulation with synthetic peptides derived from viral antigens. This assay allows the detection of memory T cells directed against the viral spike (S) protein used for vaccination as well as against other viral proteins. This method can be applied to the analysis of the immune response in individuals after covid-19 and SARS-CoV-2 vaccination and may thus be used to determine the immune preparedness of emergency workers.
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ELISpot methodology and prediction of acute rejection after renal transplantation.
Rybáková, Kateřina ; Slavčev, Antonij (advisor) ; Mrázek, František (referee)
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...
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Immune reactions induced by SARS-CoV-2 infection
Krausová, Kateřina ; Šmahel, Michal (advisor) ; Šroller, Vojtěch (referee)
Coronavirus disease 2019 (COVID-19) pandemic caused by newly discovered Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes severe health and economic problems all over the world. The disease severity depends mainly on the host's immune response to SARS-CoV-2. This virus uses many mechanisms for escape from the host's immune system. The major evasion mechanisms include suppression of interferon production at the early phase of infection, exhaustion of natural killer cells and induction of a cytokine storm. After the innate immune response, mechanisms of adaptive immunity join the defense against the virus. Patients with severe cases have a significant reduction in the amount of both helper CD4+ T cells and cytotoxic CD8+ T cells. On the contrary, these patients have an increased level of antibodies. Even though there have been many findings about immune reactions to SARS-CoV-2 in the year after its discovery, there are still many unknowns. Vaccines, which are successful at preventing COVID-19, have been developed in a short time. However, an important remaining question for further research is the longevity of immune memory after vaccination or after suffering from COVID-19.
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Donor specific T cell alloreactivity in kidney transplant recipients with borderline changes
Šilhová, Markéta ; Hrubá, Petra (advisor) ; Holáň, Vladimír (referee)
After kidney transplantation the recipient's immune system responds to the donor's antigens and the graft rejection occurs. Borderline changes are a frequent diagnosis after kidney transplantation, representing only mild rejection signs. Some patients with borderline changes undergo progression to rejection. The identification of these at- risk patients by biomarkers will allow enhanced treatment and help to prevent the development of rejection. The aim of my work was to verify biomarkers of rejection in patients with borderline changes. Chemokines CXCL9, CXCL10 and CCL17 in urine/serum of 40 patients with subclinical borderline changes at 3 months and in 25 patients with early borderline changes were determined by ELISA. At 3 months, the higher CXCL10 level predicted rejection with AUC=0.749, p=0.024. High levels of CXL10 had also been found in patients with BKV infection. We did not confirm the relationship between rejection and the CXCL9 and CCL17. In the early posttransplant period the levels of CXCL10 and CXCL9 were elevated in all patients and therefore couldn't be used to predict rejection. The alloreactivity was examined using IFN-γ ELISPOT (n=38). No association between the frequency of IFN-γ producing cells after stimulation with donor cells or CMV peptides and the development of...
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The role of B cells in transplantation reactions
Brožová, Jitka ; Slavčev, Antonij (advisor) ; Stříž, Ilja (referee)
Kidney transplantation is the best treatment for patients with end-stage renal failure. The main problem of kidney transplantation is however the development of a cellular and antibody-mediated (humoral) rejection. During the last decade, thanks to the advanced immunosuppression, prognosis of survival and function of transplanted organs has significantly improved. Nevertheless, humoral rejection remains very serious obstacle in high-risk patients, because it can permanently damage the graft. Therefore, before transplantation it is necessary to stratify patients into high and low risk groups for development of antibody-mediated rejection. Current immunogenetic tests performed before transplantation include, in addition to HLA typing, detection of panel-reactive antibodies. However, this test does not provide information about B cells which participate in the humoral response of the kidney recipient. Therefore, in the presented thesis we studied B cell reactivity and its regulation in transplanted patients. In this retrospective analysis we measured levels of the B cell activating factor, a cytokine regulating the function of B lymphocytes (BAFF). Current reports suggest that BAFF could serve as a marker of humoral rejection. Furthermore, we focused on B lymphocytes and their capacity to produce...
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ELISpot methodology and prediction of acute rejection after renal transplantation.
Rybáková, Kateřina ; Slavčev, Antonij (advisor) ; Mrázek, František (referee)
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...
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