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Changes of gut microbiome in patients with inflammatory bowel diseases
Schierová, Dagmar ; Jirásková Zákostelská, Zuzana (advisor) ; Hrdý, Jiří (referee) ; Kohout, Pavel (referee)
(EN) Microbes have coevolved with humans forming symbiotic communities that constantly challenge the immune system and, when imbalanced, could lead to diseases like inflammatory bowel disease (IBD). Patients with IBD suffer from microbial dysbiosis and chronic inflammation which could be potentiated by immune system reaction to the commensal microbiota. In the research presented here, firstly I have focused on the description of the gut and skin microbiome from patients with IBD and secondly, I investigated the process of antimicrobial defense. Patients with IBD on two different biological therapies targeting TNFα, IL-12 and IL-23 cytokines were tracked for changes in their gut and skin microbiome features. Although, neither differences in gut microbial diversity nor composition were linked with the progression of the therapies, an increased similarity to the healthy control group at week 38 of anti-TNFα therapy was found. This shift in microbiome could be considered beneficial and could be attributed to the inflammation reducing effect of the therapy. While analyzing the microbiome features, various patient characteristics were taken into account and the sources microbiome of variability were uncovered, out of which the interindividual variability stood out the most. Regarding the skin microbiome,...
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Effect of TNF-α inhibitors on microbiota composition and immune response in patients with inflammatory bowel disease
Mihula, Martin ; Jirásková Zákostelská, Zuzana (advisor) ; Grobárová, Valéria (referee)
One of the most common used therapies in inflammatory bowel diseases (IBD) treatment are inhibitors of a cytokine TNF-α. Nevertheless, up to one third of IBD patients stop respond to this therapy for unknown reason. In these days, there are not any ideal biomarkers which could predict patient's long- term response to anti-TNF-α therapy. Because the gut microbiota composition changes are tightly related to the pathogenesis of IBD, my aim in this thesis was to find out if these changes in composition are happening also due the therapy by inhibitors of TNF-α as well. Moreover, I tried to find out if there are changes in production of serum biomarkers related to the gut barrier damage and to the immune response associated with microbial translocation. Also, I focused on the immune response of IBD patients against common gut commensal bacterial antigens during the anti-TNF-α therapy. In our study, we collected for these purposes stool or blood samples from 46 IBD patients before the therapy and at 38th week from the start of the therapy and 39 healthy controls. I found that IBD patients had higher bacterial diversity (α-diversity) as well as different bacterial composition across observed groups (β-diversity) at 38th week of the anti-TNF-α therapy than before the therapy. When I divided IBD patients...
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Analysis of the anti-inflammatory effects of the bacterial components on macrophage and epithelial cell lines
Zákostelská, Zuzana
Anti-inflammatory effects of bacterial components tested on RAW 264.7, J774.A1 macrophage cell lines and on macrophages isolated from the peritoneum of BALB/c mice. Inflammatory bowel diseases (IBD) including Crohn's disease and ulcerative colitis results from a dysregulated inflammatory response of the host to intestinal microbes in genetically predisposed individuals. Despite intensively proceeding research the mechanism of this action remains unclear. In our previous studies we confirmed that some bacterial lysates isolated from Lactobacillus casei DN11400, Bacteroides distasonis and mycobacterial heat shock proteins (HSP) mitigate the severity of experimental colitis in mice. The aim of our study was to investigate whether these bacterial components have an influence on macrophages which play an important role in mediating chronic inflammation. We tested the effect of bacterial components on macrophage cell lines RAW 264.7 and J774.A1 and on macrophages isolated from the peritoneum of BALB/c mice. Viability of the macrophages we tested by flow cytometry. Qualitative and quantitative determination of cytokines in supernatants was evaluated by protein microarrays and by ELISA. Another aim was to provide evidence of changes in the activation of the NFқB signaling pathway. We observed that the bacterial...
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The Role of Microbiota in the Pathogenesis of Psoriasis
Stehlíková, Zuzana ; Jirásková Zákostelská, Zuzana (advisor) ; Demnerová, Kateřina (referee) ; Hrdý, Jiří (referee)
Psoriasis is a chronic, immune-mediated inflammatory skin disease. Its pathogenesis is associated with dysregulated cooperation among keratinocytes, innate and adaptive immune cells, coupled with environmental triggers, including microbiota. The aim of our study was to describe the microbiota composition in psoriasis and explore the role of bacteria and fungi in the pathogenesis of this disease. We used a mouse model of psoriasis induced by topical application of imiquimod (IISI) in both germ-free (GF) mice and conventional (CV) mice with microbiota manipulated by administration of a mixture of broad-spectrum antibiotics (ATB). ATB treatment markedly changed the intestinal but not the skin bacterial diversity and led to higher resistance to IISI in CV mice. Metronidazole was the most effective antibiotic, alleviating IISI symptoms in CV, but not in GF mice. This confirms that the effect of metronidazole on IISI was microbiota- dependent. Additionally, we characterized the microbiota composition of psoriatic lesions and unaffected skin in psoriatic patients compared to healthy controls, as well as the impact of different sampling approaches on uncovering cutaneous microbiota composition. We observed significant differences in α- and β-diversities when comparing identical samples sequenced on V1V2...
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Potential role of skin microbiota in the pathogenesis of dermatological diseases
Mihula, Martin ; Jirásková Zákostelská, Zuzana (advisor) ; Grobárová, Valéria (referee)
The surface of the human body is colonized by a large number of microorganisms whose composition depends not only on external and internal factors, but is also significantly influenced by the topography of human skin. The complex skin microbiota is an essential part of physiological and protective mechanisms of the skin. The change in the dynamics of microbial communities on the skin or in the gastrointestinal tract is currently considered to be part of triggering mechanisms of many skin diseases. Some of the skin inflammatory diseases are directly associated with a shift of skin microbiota composition - for instance atopic dermatitis, acne vulgaris or psoriasis. Gaining and perceiving knowledge about interspecies interactions and their effect on a host could lead to the development of new diagnostic and therapeutic approaches which could make the prevention or treatment of some skin disorders more effective. Key words: skin, skin diseases, skin microbiota, immunity, psoriasis, atopic dermatitis, vitiligo
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Composition of skin microbiome in psoriatic patients
Stehlíková, Zuzana ; Jůzlová, P. ; Rob, F. ; Herzogová, J. ; Koren, O. ; Uzan, A. ; Tlaskalová-Hogenová, Helena ; Jirásková Zákostelská, Zuzana
Psoriasis is a chronic noninfectious and inflammatory skin disease, whose pathogenesis involves environmental triggers, including microbiota. In our study we compared bacterial composition between healthy controls and psoriatic patients using V1V2 hypervariable region of 16S rRNA. We found higher species diversity in psoriatic lesions than in contralateral psoriatic healthy site or in healthy control skin. Genus Propionibacterium was more abundant in contralateral healthy sites (57.77%) than in affected psoriatic sites of psoriatic patients (47.12%). On the other hand, we found higher abundance of genus Staphylococcus in psoriatic lesions (18.78%), while lower abundance in contralateral healthy sites (9.52%). Since Propionibacterium are commensal bacteria, the shift in their abundance from healthy to psoriatic skin could be due to disturbation of natural skin habitat. Interpretation of higher presence of Staphylococcus in psoriatic lesions comparing to contralateral healthy sites requires further species characterization. To complete the picture about psoriatic microbiome we will further investigate skin fungal composition in identical samples.
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Role of microbiota in mouse experimental model of psoriasis
Jirásková Zákostelská, Zuzana ; Stehlíková, Zuzana ; Klimešová, Klára ; Rossmann, Pavel ; Dvořák, Jiří ; Novosádová, Iva ; Kostovčík, Martin ; Coufal, Štěpán ; Šrůtková, Dagmar ; Hudcovic, Tomáš ; Štěpánková, Renata ; Rob, F. ; Jůzlová, P. ; Herzogová, J. ; Tlaskalová-Hogenová, Helena ; Kverka, Miloslav
Anotace v anglickém jazyce\n\nMouse model of human psoriasis and gnotobiotic are important tools in understanding the role of gut and skin microbiota in pathogenesis of psoriasis. In our experiments we showed that gnotobiotic mice, as well as conventional mice treated with antibiotics, have milder skin inflammation in comparison with control conventional mice. Treatment with broad spectrum antibiotics led to dramatic shift in gut microbial composition, in particular, we observed extensive increase of order Lactobacillales. To analyze the potential effect of Lactobacillales on skin inflammation, we further monocolonized mice with L. plantarum WCFS1. Also monocolonized mice showed lower skin inflammation in comparison with conventional mice. To understand whether microbial dysbiosis is cause or effect of psoriasis needs to be further investigated.\n\n
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The effects of bacterial lysates on the gut barrier function and microbiota composition
Zákostelská, Zuzana ; Tlaskalová - Hogenová, Helena (advisor) ; Prokešová, Ludmila (referee) ; Rada, Vojtěch (referee)
Dynamic molecular interactions between the microbiota and the intestinal mucosa play an important role in the establishment and maintenance of mucosal homeostasis. Aberrant host- microbiota interaction could lead to many diseases such as inflammatory bowel disease. The aim of our study was to evaluate the commensal and probiotic bacteria activities and their ability to induce pathological or exert beneficial effects. The most important trigger for immune system development is an exposure to microbial components. Here, we show that there is a time window at about three weeks of age, which enables the artificial colonization of germ free mice by a single oral dose of cecal content. The delayed colonization by either inoculation or co-housing causes permanent changes in immune system reactivity, which may downgrade the results of experiments performed on first generation of colonized animals. In this thesis we report that even non-living commensal bacteria such as Parabacteroides distasonis (mPd) or well known probiotics such as L. casei DN-114 001 (Lc) possess anti-inflammatory effects in experimental model of colitis. The mechanisms that this effect is achieved by the lysate of L. casei DN-114 001 comprise: a) improvement in the gut barrier function, b) correction of the dysbiosis, and c) modulation of the...
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Analysis of the anti-inflammatory effects of the bacterial components on macrophage and epithelial cell lines
Zákostelská, Zuzana
Anti-inflammatory effects of bacterial components tested on RAW 264.7, J774.A1 macrophage cell lines and on macrophages isolated from the peritoneum of BALB/c mice. Inflammatory bowel diseases (IBD) including Crohn's disease and ulcerative colitis results from a dysregulated inflammatory response of the host to intestinal microbes in genetically predisposed individuals. Despite intensively proceeding research the mechanism of this action remains unclear. In our previous studies we confirmed that some bacterial lysates isolated from Lactobacillus casei DN11400, Bacteroides distasonis and mycobacterial heat shock proteins (HSP) mitigate the severity of experimental colitis in mice. The aim of our study was to investigate whether these bacterial components have an influence on macrophages which play an important role in mediating chronic inflammation. We tested the effect of bacterial components on macrophage cell lines RAW 264.7 and J774.A1 and on macrophages isolated from the peritoneum of BALB/c mice. Viability of the macrophages we tested by flow cytometry. Qualitative and quantitative determination of cytokines in supernatants was evaluated by protein microarrays and by ELISA. Another aim was to provide evidence of changes in the activation of the NFқB signaling pathway. We observed that the bacterial...
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