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Characterization and identification of wheat allergens from foodstuffs containing wheat
Goliáš, Jaroslav ; Prokešová, Ludmila (referee) ; Tučková, Ludmila (advisor)
In some cases, ordinarily accepted food can evoke harmful clinical symptoms which are brought on by sensitivity on specific component of foodstuff. This harmful reaction on accepted food is called food allergy and it tends to go up in the world's population in last years. The food allergy affects especially children as well as adults. The important role in the food allergy plays wheat and foodstuff containing components of wheat. Wheat allergens from flour is already well-known, but man accepts wheat in food in the treated form (heat treated food) what can decrease or increase allergenicity. Moreover, enzymes presented in gastrointestinal tract of man can influence allergenicity of wheat allergens too. The goal of this thesis is characterization of allergens presented in heat treated wheat foodstuff and in wheat foodstuff digested by enzymes in gastrointestinal tract of man. Characterization of these altered and newly produced allergens can help find better diagnostic techniques, increase specificity of detection of IgE antibodies, enable pointed therapy of allergic patients or find and apply new and suitable diet.
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Immune response of mouse respiratory tract after immunization with influenza virus
Berková, Veronika ; Tučková, Ludmila (referee) ; Prokešová, Ludmila (advisor)
Immune response of mice after mucosal immunization by influenza virus type A with bacterial adjuvant Bacillus firmus Immunization of mice by inactivated influenza virus via respiratory tract induces a good mucosal and systemic immune response if bacterial adjuvant - delipidated G+ non- pathogenic bacterium Baccillus firmus (DBF) - is used. BALB/c mice were immunized intratracheally (IT) or intranasally (IN) with inactivated influenza A/PR/8/34 virus in combination with adjuvant DBF (50, 100, 200 or 500μg per immunization dose). We tested the production of antibodies against homologous virus and cross-reacting antibodies against subtype H3N2, H6N2 and H9N2 viruses in serum and mucosal secretions of nose, lungs and intestine by the ELISA method. Immunization of mice with virus itself induces the production of antibodies against homologous virus and lower production of cross-reacting antibodies against heterologous subtypes. Immunostimulatory adjuvant activity (optimal 100μg per immunization dose) enhances systemic and mucosal antibody production against homologous virus (H1N1) and markedly against heterologous subtypes (H3N2, H6N2, H9N2), especially after IT immunization of mice. For evaluation of cellular immunity, we tested spleen cell proliferation of immunized mice by 3 H-thymidine incorporation and...
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Adjuvant Mucosal Immunization of Mice against inFluenza Virus
Zanvit, Peter ; Prokešová, Ludmila (advisor) ; Tlaskalová - Hogenová, Helena (referee) ; Kopecký, Jan (referee)
Influenza virus infections cause considerable morbidity and mortality in the world. Current immunization against influenza is provided using parenterally given influenza vaccines. These vaccines can induce good systemic immunity but they fail to induce a protective mucosal immunity. Because of persisting threat of new highly pathogenic influenza A subtypes development, a vaccination inducing intersubtypic cross-protection is desirable. The principal aims of the present study were: firstly, to explore the effect of Bacillus firmus (BF) and its delipidated form (DBF) as mucosal adjuvants for immunization via respiratory tract, secondly to test the potencial of BF and DBF to induce intrasubtypic and intersubtypic protection and finally to characterize the mechanism of adjuvant effect. (...) The mechanism of adjuvant effect was tested in NALT after intranasal immunization of mice by inactivated influenza virus type A, adjuvant alone (DBF) and by mixture of virus+DBF. We tested the expression of selected genes for cytokines, toll-like receptors and other genes participating in immune response by qPCR. Intranasally given DBF and mainly mixture virus+DBF induced expression of cytokines characteristic for Th1 immune response (IFN-and IL-2) whereas expression of genes characteristic for Th2 was decreased (IL-4)....
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Participation of some mechanisms of mucosal immunity in the pathogenesis of Crohn's disease and ulcerative colitis
Drastich, Pavel ; Špičák, Julius (advisor) ; Prokešová, Ludmila (referee) ; Nečas, Emanuel (referee)
In our study we contributed to clarifying the important role of mucosal immunity system in the pathogenesis of inflammatory bowel disease. We have shown that upregulated expression of TLR2, TLR4 and CD14 in patients with Crohn disease (CD) and ulcerative colitis (UC) reflects the consequences of the interaction of intestinal epithelial cells and macrophages in the lamina propria with intestinal microbiota. TLR2 expression in the terminal ileum is significantly increased regardless of whether assessed in active or remission stage of UC. Possible explanation of this unexpected finding could be that this upregulation reflects the activation of innate immunity cells by as yet unknown bacterial components also present in the proximal part of the gut of UC patients. Thus, it seems that the terminal ileum can sensitively react to "potentially pathogenic" but still undiscovered components of microbiota by immological mechanisms impairing the homeostasis in colonic tissue. Furthermore we showed that the mucosal interleukin- 6 (IL-6) could be expressed and produced in different parts of the intestine with different intensity. Unexpected high IL-6 mucosal level in the rectal mucosa without any signs of macroscopic inflammation could reflect the susceptibility of this area to perianal and perirectal disease and fistula...
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Antimicrobial lytic factors of the coelomic fluid of Earthworms
Josková, Radka ; Bilej, Martin (advisor) ; Horák, Petr (referee) ; Prokešová, Ludmila (referee)
I. SUMMARY lnvertebratesare widely distrbuted animals.They can be found in almostany krnd ofhabitat. Their successfulsurvival shategiesare basedon short life span combinedwith numerousoffspring and, more importantly,all invertebratespecieshavedevelopeda wriety ofdefensemechanismselficientlyrecognizingand respondingto non-selfsubstances. The defensemechadsmsof earthwoms were studiedover the pastfou decades.It becameappdent thattheearthworms.aswell asotherinvertebrates,lackspecificimmunoglobulins,lymphoc)4esor otherfeatures ofthe adaptiveimmunesystemdescribedin verteblates,but possessirmatedefensecomponents In this thesis,we foousedon the detaildescriptionof somedefensemoleculesinvolvedin imate immunityof earthworms. l. Lysozyme is an erzyme with strong antibacterialactivity describedin many organisms. We characterizedthelysozymeof Erieniaandrei(formerly E.fetida andrei)earthwormbothstruchmlly and functionally.Moleoularcharactefiationof lyso4rne providesa new tool for monitoringof innateimmunity in earthworms. 2. A cltol''tic effect of the coelomicfluid of E. fetida was observedin experimentswith TNF-sensitive tumorL929 cell line. Subsequentisolationof l)tic proteinsled to theidentificationof 42-kDaprotein,which wasnamedcoelomiccytolyticfactor- CCF.CCFwasshom to bepresentalsoin coelomicfluid of mother...
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The role of heat shock proteins in pathogenesis of leukaemia, graft versus host disease, juvenile idiopathic and rheumatoid arthritis"
Sedláčková, Lucie ; Hromadníková, Ilona (advisor) ; Říhová, Blanka (referee) ; Prokešová, Ludmila (referee)
The aims of the study: Heat shock proteins (Hsps) represent major immunodominant antigens in a wide spectrum of microbial pathogens and immune reactivity against different members of HSP families accompanies many infectious diseases. Known role of Hsps in antigen resentation could also suggest their potential role in the alloreactive process that leads to acute graft versus host disease (GvHD). sequence homology and cross-reactivity between microbial and human Hsps led to the concept that Hsps might be involved in the atiopathogenesis of autoimmune diseases such as juvenile idiopathic arthritis (JIA) and rheumatoid arthritis (RA). The major groups of molecular chaperones have been implicated in cancer development as well and stress-inducible Hsp70 was found to be present on the plasma membrane of tumour cells but not on the corresponding normal tissues. On the basis of previous observations we focused our study on the role of Hsps, namely Hsp60 and Hsp70, in the pathogenesis of leukaemia, graft versus host disease, juvenil idiopathic arthritis and rheumatoid arthritis. Powered by TCPDF (www.tcpdf.org)
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Role of bacteria and mucosal immune system and their interaction in the pathogenesis of inflammatory bowel disease
Du, Zhengyu ; Hudcovic, Tomáš (advisor) ; Prokešová, Ludmila (referee) ; Kamanová, Jana (referee)
Although the etiology and pathogenesis of inflammatory bowel disease (IBD) is not fully understood, it is generally accepted that the inflammation results from aberrant immune responses to antigens of gut microbiota in genetically susceptible individuals (Sartor et al., 2006). Alteration in intestinal microbiota has been found in IBD patients with increased abundance of certain bacteria and decreased abundance of others. Due to the complexity of the disease, multifaceted interactions between genetic factors, host immune response, gut microbiota and environment factors need to be taken into account. In this thesis, the pathogenesis of IBD was first reviewed in respect with the four factors mentioned above. Then we concentrated on the interaction between IBD-associated bacteria and mucosal immune system. We investigated the ability of mucosal-associated bacteria (MAB) from IBD patients to induce spontaneous colitis in germ-free (GF) mice and the impact of those bacteria on the development of dextran sulfate sodium (DSS)-colitis. Together with the analysis of the composition of gut microbiota of MAB colonized mice, we demonstrated the potential deleterious microbes were able to increase the susceptibility to DSS-colitis once they found a suitable niche. We revealed the mechanism of an E.coli strain...
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