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Monitoring of sublingual allergen immunotherapy using serum cytokine levels
Brzáková, Adéla ; Řezáčová, Vladimíra (advisor) ; Andrýs, Ctirad (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Bc. Adéla Brzáková Supervisor: Mgr. Vladimíra Řezáčová, Ph.D. Title of thesis: Monitoring of sublingual allergen immunotherapy using serum cytokine levels Keywords: allergen immunotherapy, cytokine, ELISA, grass pollen allergy Background: The aim of this thesis was to find out whether there are detectable changes in the levels of IFNγ, IL-4 and IL-5 during allergen immunotherapy and whether these cytokines could thus serve as biomarkers of the efficiency of this treatment. Methods: Selected cytokines were determined using Quantikine® HS ELISA Human IFN-γ Immunoassay, Quantikine® HS ELISA Human IL-4 Immunoassay and Quantikine® ELISA Human IL-5 Immunoassay. The monitored population consisted of 32 polinotics, 19 of which were administered allergen immunotherapy and 13 of which constituted the control group. As part of the follow-up, both groups also continuously recorded the severity of their symptoms using a visual analog scale. Results: In the group of treated patients, a decrease in IL-4, IL-5 and IFNγ concentrations was observed during treatment, but no significant differences in the values of these cytokines were found between patients treated with Oralair and control individuals. Using...
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Comparison of two multiplex (biochip) methods for determination of specific IgE against molecular defined allergens
Velčovská, Denisa ; Andrýs, Ctirad (advisor) ; Jankovičová, Karolína (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Subject of study: Bioanalytical laboratory diagnostics in health care Author: Bc. Denisa Velčovská Supervisor: prof. RNDr. Ctirad Andrýs, Ph.D. Title of diploma thesis: Comparison of two multiplex (biochip) methods for determination of specific IgE antibodies against food allergens in group of patients suffering from atopic dermatitis This thesis deals with the comparison of two multiplex (biochip) methods ALEX and ISAC in the determination of specific IgE antibodies against selected allergens in patients with atopic dermatitis. The aim of this thesis is to statistically compare the two methods and perform an evaluation of the data obtained, to assess whether the two methods provide similar results and are interchangeable, or whether a statistically significant difference can be observed. This diploma thesis mainly focuses on inhalant allergens of moulds, mites, house dust mites and animal allergens. The theoretical part of this diploma thesis is focused on allergies and their clinical forms. It also describes allergens, their nomenclature, biochemical structure and methods used to diagnose allergic inflammation. In the experimental part, the basic statistics of the cohort, which includes 96...
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Monitoring of sublingual allergen immunotherapy using serum cytokine levels
Brzáková, Adéla ; Řezáčová, Vladimíra (advisor) ; Andrýs, Ctirad (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Bc. Adéla Brzáková Supervisor: Mgr. Vladimíra Řezáčová, Ph.D. Title of thesis: Monitoring of sublingual allergen immunotherapy using serum cytokine levels Keywords: allergen immunotherapy, cytokine, ELISA, grass pollen allergy Background: The aim of this thesis was to find out whether there are detectable changes in the levels of IFNγ, IL-4 and IL-5 during allergen immunotherapy and whether these cytokines could thus serve as biomarkers of the efficiency of this treatment. Methods: Selected cytokines were determined using Quantikine® HS ELISA Human IFN-γ Immunoassay, Quantikine® HS ELISA Human IL-4 Immunoassay and Quantikine® ELISA Human IL-5 Immunoassay. The monitored population consisted of 32 polinotics, 19 of which were administered allergen immunotherapy and 13 of which constituted the control group. As part of the follow-up, both groups also continuously recorded the severity of their symptoms using a visual analog scale. Results: In the group of treated patients, a decrease in IL-4, IL-5 and IFNγ concentrations was observed during treatment, but no significant differences in the values of these cytokines were found between patients treated with Oralair and control individuals. Using...
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Bazotests and lymphocyte transformation tests linked to drug intolerance
Honsárková, Tereza ; Krčmová, Irena (advisor) ; Andrýs, Ctirad (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Science Candidate: Bc. Tereza Honsárková Supervisor: prim. MUDr. Inrena Krčmová, CSc. Title of diploma thesis: Bazotests and lymphocyte transformation tests linked to drug intolerance Basophil activation tests and lymphocyte transformation tests are methods that allow drug allergy diagnosis. The right diagnosis is crucial for treatment. Drug allergy is often associated with β-lactam antibiotics hypersensitivity and especially with penicillins. The aim of this diploma thesis is to provide actual statistics based on selected patient medical reports. Those patients were completely clinically and laboratory examined. The main condition for data selection was the execution of skin tests for suspicion of drug allergy caused by penicillin-like antibiotics at the Institute of Clinical Immunology and Allergology of the Faculty Hospital Hradec Králové in 2017. Patients' anamnesis, skin tests and laboratory examination focused on basophil activation tests and lymphocyte transformation was composed. Statistics of 25 patients shows beneficial informations. Delayed manifestations are the most prevalent (63 %) in patients which is interesting. However the patients with positive skin test on major determinant had acute...
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Immunological laboratory diagnostics of non-specific gut inflammation.
Vohralíková, Jana ; Jílek, Petr (advisor) ; Andrýs, Ctirad (referee)
BACKGROUND: Inflammatory bowel disease (IBD), with Crohn‛s Disease (CD) and ulceratice colitis (UC) as the two main disorders, is a heterogenous group of disease of unknown etiology. IBD serological markers may be important in the diagnosis of IBD, for differentiating CD from UC. The major serological marker in commonal use are ASCA and ANCA. Recently new antiglycan antibodies have been identified: anti laminaribioside carbohydrate antibodies - ALCA and anti chitobioside carbohydrate antibodies ACCA. The antibodies are significantly associated with CD. AIMS: To evaluate the significance of the recently discovered novel antibodies: ALCA and ACCA METHODS: Assesment of pANCA, ASCA, ALCA and ACCA was performed using the standardized indirect immunoflourescence technique and ELISA. Serum samples were obtained from 89 pacients with CN and from 33 pacients with UC. Results were considered positive if: ASCA> 15 U/ml, ALCA> 60 EU and ACCA> 90 EU. RESULTS: ASCA was positive for 78.8% (70/89) of patiens with CD and 12,1% (4/33) of pacients with UC. pANCA was positive for 42.4%(14/33) of patiens with UC and 7,8%(7/89) of patiens with CN. The ASCA IgA test yielded 71.3% sensitivity and 93.9% specificity for CN. The ASCA IgG test yielded 57,3% sensitivity and 93.9% specificity for CN. The ANCA test yielded 41.9%...
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Cholesterol depletion using reopheretic and apheretic processes and impact of these methods on immune system
Svrčková, Ellen ; Andrýs, Ctirad (advisor) ; Krejsek, Jan (referee)
LDL-apheresis and haemorheopheresis are the most frequent methods of extracorporeal ellimination methods used for lowering the LDL cholesterol in patients with familial hypercholesterolaemia (FH). In case of failure of conservative therapy (represented by pharmacotherapy and/or dietary regime) these methods reprensent the effective and accessible solution for the clinical status characterized by high morbidity and mortality as well. The lipid components are the most frequent observed markers for the effeciveness of intervention. However, immunity with its effector molecules plays also essential role and there is suppose, that could also reflect the state and progress of atherosclerosis in FH patients. The aim of this thesis was to evaluate the levels of selected immunological markers (plasmatic glykoprotein α-2-macroglobulin, IL-10, soluble endoglin, soluble apoptotic factor sAPO-Fas and soluble form of adhesive P-selectin) and their changes after LDL-apheresis and haemorheopheresis employing enzyme immunoanalysis and immuno nephelometry. Totally, 12 patients were involved in this study, 3 were treated by haemorheopheresis, the rest 9 received LDL-apheresis. As a results, significant decreases in serum levels of α-2- macroglobulin, soluble endoglin and sAPO-1/Fas were recorded. Observed changes of...
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The sensitivity of C-reactive protein levels after total hip endoprosthesis
Fremundová, Petra ; Jílek, Petr (advisor) ; Andrýs, Ctirad (referee)
C-reactive protein is an acute phase protein and is one of the longest used laboratory methods in the diagnosis of infectious or other inflammatory conditions. Replacement of the hip joint is one of the most common orthopedic surgeries performed on workplaces. One of the most serious complications of these procedures is an infection in the area of compensation. This work deals with the diagnostic possibilities of prevention of infectious complications after replacement of the hip joint using measurements of the levels of C-reactive protein in the postoperative period. On the set of 30 patients after replacement of the hip joint was monitored the development of the values of C- reactive protein at regular intervals in the postoperative period. Levels were set one of the available optical methods and evaluated 1., 3., 5. and 7. of post-operative day. On the basis of the results obtained with the work of trying to evaluate their own sensitivity and specificity of laboratory tests in C - reactive protein in the early postoperative period after the replacement of the hip joint. From the results, it is obvious a significantly higher sensitivity tests than its specificity. At the same time, however, is a clear positive contribution to the examination of the dynamics, in particular in monitoring the levels...
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Pentraxin PTX3 - a new marker of immunopathological inflammation.
Kondělková, Kateřina ; Jílek, Petr (advisor) ; Andrýs, Ctirad (referee)
Pentraxin 3 (PTX3) is a newly identified acute phase reactant which shares some structural and some functional properties with CRP, classical short pentraxin. On the other hand, PTX3 displays unique biological properties of its own, including a possible role in the pathogenesis of cardiovascular diseases and in processes accompanying the natural evolution of surgical wounds. Unlike CRP, which is manufactured predominatly in the liver, PTX3 is produced especially at sites of tissue damage. PTX3 synthesis is induced by proinflammatory cytokines TNF-α or IL-1β and by microbial wall constituents such as the lipololysaccharide (LPS). Plasma protein PTX3 concentrations are elevated in a wide range of diseased states, such as in coronary artery disease, in pulmonary infection and acute lung injury, 3 in patients with chronic kidney disease, in pacients with depression, during normal pregnancy and preeclampsia, in pacients with psoriasis and during cardiac surgery with or without cardiopulmonary bypass. PTX3 was detected using detection set (Alexis Biochemicals, Switzerland) cat.no. ALX- 850-299-KI01 for sandwich ELISA application that provided capture monoclonal antibody to PTX3, detection polyclonal antibody to PTX3 and recombinant PTX3 (standard). Goeckerman's therapy of psoriasis is highly efficient in...
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The effect of Goeckerman therapy on selected immune markers in patients with psoriasis
Kondělková, Kateřina ; Andrýs, Ctirad (advisor) ; Posová, Helena (referee)
1 Summary The effect of Goeckerman therapy on selected immune markers in patients with psoriasis Psoriasis is a systemic inflammatory disorder with skin manifestation. Classical approach to treat psoriasis is Goeckerman therapy (GT) which is based on daily application of pharmaceutical coal tar on affected skin with subsequent exposition to UV light. The aim of this study was to observe selected immunological markers and cell subpopulations in patients with psoriasis and to elucidate the effect of GT on these parameters. We evaluated T regulatory cells (Treg), membrane CD163 expression on monocytes, serum levels of CD163 (sCD163), membrane TLR2 and TLR4 expression on monocytes and serum levels of TLR2 (sTLR2). The immunofluorescent analysis was performed on erythrocyte lysed peripheral blood using a flow cytometer FC500 Cytomics (Beckman Coulter) and FACSCalibur (Becton Dickinson). The level of soluble markers was detected by sandwich enzyme-liked immunosorbent assay (ELISA). There was no significant difference in the relative number of Treg cells in the peripheral blood of healthy blood donors and patients with psoriasis before initiation of GT. The relative number of Treg is significantly increased in psoriatic patients after GT compared to the pre-treatment level. In comparison with levels in controls...
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