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The use of immunoregulatory properties of mesenchymal stem cells/ and their therapeutic potential
Javorková, Eliška
Mesenchymal stem cells (MSCs) have the potential to differentiate into various cell types, possess potent immunomodulatory properties and can influence various functions of immune cells. Since the immunomodulatory properties of MSCs can be modified by cytokines, we compered the effect of unstimulated MSCs and MSCs pretreated with interleukin (IL)-1, interferon (IFN)- , transforming growth factor (TGF)- and IL-10 on the development of regulatory T cells (Treg) and T helper 17 (Th17) cells in vitro and on the inflammatory environment in the eye. MSCs can produce significant levels of TGF- and IL-6. These cytokines represent the key factors that reciprocally regulate the development of naive T cells into Treg and Th17 cells. Unstimulated MSCs produce TGF- , but not IL-6, and the production of TGF- can be further enhanced by IL-10 or TGF- . In the presence of IL-1, MSCs secrete significant levels of IL-6, in addition to spontaneous production of TGF- . MSC producing TGF- induced preferentially expression of Foxp3 and activation of Treg lymphocytes, whereas MSCs supernatants containing TGF- together with IL-6 supported ROR t expression and development of Th17 cells. We demonstrated that MSCs and their products effectively control the development of Tregs and Th17 cells in a population of...
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Use of the nanofiber scaffold for transfer of stem cells onto the injured ocular surface in mouse experimental model
Kössl, Jan ; Zajícová, Alena ; Heřmánková, Barbora ; Javorková, Eliška ; Boháčová, Pavla ; Holáň, Vladimír
Corneal damage is one of the most common causes of impaired vision or even blindness. When the injury is more extensive and the limbal region is involved, the natural regeneration of the cornea is not sufficient. Such damage can lead to the limbal stem cell deficiency (LSCD). The only option for LSCD treatment is transplantation of the limbal tissue or a transfer of limbal stem cells (LSCs) cultured from the healthy eye. The allogenic transplantation of the limbus or cultivated LSCs with a systemic administration of immunosuppressive drugs is needed in the case of bilateral LSCD. Nevertheless, the cell therapy is very promising approach for LSCD treatment. Transplantation of mesenchymal stem cells (MSCs) seeded on an appropriate scaffold turned out to be a suitable therapy of the LSCD. In our experimental model of LSCD we use nanofiber scaffold for MSC and LSC cultivation and for transplantation of these cells onto the chemically injured mouse eye. MSCs have immunosuppressive and immunomodulatory properties. We showed that MSCs have the ability to inhibit production of molecules associated with the inflammation and support epithelial regeneration in the damaged cornea. These inhibitory properties were confirmed in both in vitro and in vivo mouse model. Results thus showed beneficial effects of stem cell transplantation for murine corneal healing and for suppression of a local immune reaction which can impede the healing process. Such similarity of in vivo and in vitro results allows us further experiments to clarify mechanisms of MSC regenerative and healing properties after the transplantation onto the injured cornea.
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The use of immunoregulatory properties of mesenchymal stem cells/ and their therapeutic potential
Javorková, Eliška
Mesenchymal stem cells (MSCs) have the potential to differentiate into various cell types, possess potent immunomodulatory properties and can influence various functions of immune cells. Since the immunomodulatory properties of MSCs can be modified by cytokines, we compered the effect of unstimulated MSCs and MSCs pretreated with interleukin (IL)-1, interferon (IFN)- , transforming growth factor (TGF)- and IL-10 on the development of regulatory T cells (Treg) and T helper 17 (Th17) cells in vitro and on the inflammatory environment in the eye. MSCs can produce significant levels of TGF- and IL-6. These cytokines represent the key factors that reciprocally regulate the development of naive T cells into Treg and Th17 cells. Unstimulated MSCs produce TGF- , but not IL-6, and the production of TGF- can be further enhanced by IL-10 or TGF- . In the presence of IL-1, MSCs secrete significant levels of IL-6, in addition to spontaneous production of TGF- . MSC producing TGF- induced preferentially expression of Foxp3 and activation of Treg lymphocytes, whereas MSCs supernatants containing TGF- together with IL-6 supported ROR t expression and development of Th17 cells. We demonstrated that MSCs and their products effectively control the development of Tregs and Th17 cells in a population of...
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GENE EXPRESSION AND IMMUNOLOGICAL RESPONSE IN MICE EXPOSED TO ZnO NANOPARTICLES
Rössner ml., Pavel ; Vrbová, Kristýna ; Strapáčová, S. ; Rössnerová, Andrea ; Ambrož, Antonín ; Brzicová, Táňa ; Líbalová, Helena ; Javorková, Eliška ; Zajícová, Alena ; Holáň, Vladimír ; Kulich, P. ; Večeřa, Zbyněk ; Mikuška, Pavel ; Coufalík, Pavel ; Křůmal, Kamil ; Čapka, Lukáš ; Dočekal, Bohumil ; Šerý, Omar ; Machala, M. ; Topinka, Jan
We analyzed gene expression changes in the lungs and the immunological response in splenocytes of mice exposed by inhalation of ZnO nanoparticles - NP. Adult female ICR mice were treated for three days and three months, respectively. Analysis of differential expression in genes involved in oxidative stress was conducted using quantitative RT-PCR. The potential immunotoxic and immunomodulatory effects of ZnO NP were analyzed by phenotyping and cytokine production by splenocytes after three months exposure. Three days exposure resulted in down-regulation of GCLC, GSR, HMOX-1, NQO-1, NF-kB2, PTGS2 and TXNRD1 mRNA expression, three months exposure increased the expression of these genes. Three months exposure caused a significant decrease in the percentage of granulocytes in the spleen cells, and affected the production of IL-10 and IL-6 by lipopolysaccharide-stimulated leukocytes. In summary, our study revealed changes in the expression of genes involved in the oxidative stress response following acute ZnO NP exposure. Subchronic ZnO NP exposure induced immunomodulatory effects in the spleen.
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Immunogenic cell death
Šímová, Michaela ; Drbal, Karel (advisor) ; Javorková, Eliška (referee)
According to the danger model, the immune system is activated by endogenous molecules known as danger-associated molecular patterns (DAMP) that are externalized from the interior of a dying cell to the cell surface or released into the extracellular space. Due to the loss of plasma membrane integrity a necrotic cell death as well as several types of proinflammatory programmed cell death are considered to be immunogenic, whereas apoptosis, on contrary, has been initially defined as a tolerogenic type of cell death. However, under certain circumstances, the immune response can be initiated by an apoptotic cell after exnternalization of DAMP molecules by newly described secretory pathways. This phenomenon was observed on tumor cells as a result of some widely used therapeutic modalities and is known as immunogenic cell death (ICD). Nomenclature of selected types of cell death is part of this thesis. The aim of this bachelor thesis is to provide an evidence of the experimental support for ICD theory during in vivo initiation of the immune response. I will evaluate the correlation between ICD and the induced exposure of DAMP molecules on the surface of tumor cells or their secretion to the extracellular space.
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Modulation of human macrophages and renal epitelium chemokine profile
Pidhorodetská, Halyna ; Stříž, Ilja (advisor) ; Javorková, Eliška (referee)
One of the main effects of pro-inflammatory cytokines is the induction of chemokines and the expression of adhesive molecules that regulate the migration of immune cells to the center of the damage. Chemoattractant gradient also provides a physiological delivery of cells to tissues and lymphatic organs under normal circumstances. Chemokines are chemotactic cytokines that form a very large and diverse group of secreted proteins that have many functions both in processes that maintain homeostasis but also in inflammatory states. Production of some chemokines also has a major effect on graft rejection. Further understanding of the mechanisms involved in the acute rejection chemokine could contribute to improving treatment steps in transplantology. In this diploma thesis, serum chemokine levels were monitored in renal transplant patients, but these measurements did not show significant dynamics. Furthermore, the effect of pro-inflammatory cytokines on the release of chemokines from renal epithelial cells and monocytes was studied. Experiments were performed to monitor the levels of individual chemokines such as ENA-78, IL-8, MCP-1, MIP-1 β, RANTES, GRO alpha, THP-1 (monocyte/macrophage cell line), RPTEC (renal epithelial cells of proximal tubules) and RA (renal cell tumor lines). TNF-α (tumor necrosis...
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Different capacity of in vitro generated monocyte-derived dendritic cells of newborns of healthy and allergic mothers to prime immune responses
Súkeníková, Lenka ; Hrdý, Jiří (advisor) ; Javorková, Eliška (referee)
(EN) Reduced microbial stimulation of an immature neonatal immune system can lead to a poor balance adjustment of immune responses, thus contributing to the development of allergic diseases, whose incidence continues to rise. One of the promising precautionary measures seems to be an early preventive administration of probiotic bacteria to pregnant or nursing mothers, or to newborns. Previous works have described a beneficial effect of Escherichia coli O83:K24:H31 (E. coli O83) in the prevention of allergic diseases. In order to contribute to the clarification of E. coli O83 effects on the neonatal immune system, its immune- modulating properties were tested in vitro on umbilical cord blood cells. The ability of E. coli O83 to support the maturation of in vitro-derived dendritic cells from cord blood precursors (moDCs) of the children of healthy (children with a relatively low risk of allergy) and allergic (children at a relatively high risk of developing allergies) mothers was tracked by flow cytometry, qPCR and ELISA. Probiotic bacteria-stimulated moDCs were subsequently cultured with autologous naive CD4+ T lymphocytes and immune response polarization was also characterised by flow cytometry, qPCR, and ELISA. It was evident from the results that E. coli O83 promoted moDCs maturation. The presence of...
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Immunosuppression in the microenvironment of glioblastoma
Ternerová, Nikola ; Stollinová Šromová, Lucie (advisor) ; Javorková, Eliška (referee)
Glioblastomas (GBM) are the most malignant brain tumors, which are thought to originate from neoplastic transformation of glial cells. These tumors are characterized with highly infiltrative growth, neovascularization, and radio- and chemoresistance. In spite of current therapy including surgical resection of the tumor and chemo/radio therapy, patient's prognosis is still poor and median survival is about 15 months. Certain non-tumor cells present in the GBM microenvironment participate in tumor progression using mechanisms contributing to the local and systemic immunosuppression. Critical roles in the immune escape of GBM have the regulatory T-cells (Tregs), the tumor-associated macrophages (TAMs) and the myeloid-derived suppressor cells (MDSCs). Immunosuppressive mechanisms in GBM are conducted through direct cell-mediated contacts and soluble mediators secreted by tumor-associated cells into the local tumor microenvironment and circulating blood. Both these processes may inhibit immune response mounted against cancer cells. Certain cancer associated cells and secreted mediators are distributed by peripheral blood and potentiate systemic immunosuppression in the GBM host organism. Gaining knowledge about these mechanisms may reveal to possible targets for GBM immunotherapy. For instance,...
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IL-15/IL-15Rα complexes as IL-15 superagonist
Honzírková, Lucie ; Kovář, Marek (advisor) ; Javorková, Eliška (referee)
Interleukin 15 binds to its hight-afinity receptor IL-15Rα and forms stable IL-15/IL-15Rα complexes. IL-15Rα presents IL-15 in trans to target CD122/CD132 cells, where initiates signal transduction. Interleukin 15 has important role in immune system as it - regulates the homeostatic proliferation of memory CD8+ T cells and it is an essential for function, development and homeostasis of NK and NKT cells. Cell surface-expressed IL-15/IL15Rα complexes can be completely substituted by soluble recombinant IL-15/IL-15Rα-Fc complexes, which have substantially higher biological activity in composition to free IL-15 and behave as IL-15 superagonist. Il-15/IL-15Rα-Fc complexes are thus promising tool for some clinical use, i.e. cancer immune therapy, treatment of HIV or improvement of vaccination.
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