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Toll like receptors and myeloid cells in development and disease
Balounová, Jana ; Filipp, Dominik (advisor) ; Špíšek, Radek (referee) ; Vannucci, Luca Ernesto (referee)
Toll like receptors (TLRs) are germline-encoded pattern recognition receptors (PRRs) that play a central role in host cell recognition and responses to pathogens. Primarily they are responsible for induction and regulation of the innate and adaptive immune responses whereby the effector function is executed chiefly by differentiated myeloid cells. Somewhat unexpectedly, TLRs have been also shown to be involved in direct pathogen sensing by bone marrow-derived hematopoietic stem cells (HSCs) and hematopoietic progenitors when, under inflammatory conditions, the rapid generation of innate immune effector cells that effectively combat the infection is of utmost priority. While it has been recognized that the release of inflammatory cytokines from inflamed tissues along with the changes in proportions of differentiating cells in the bone marrow (BM) as well as the BM niche can nudge the differentiation of adult BM-derived cells towards myeloid cells and granulocytes, a direct role of TLRs expressed by HSCs in this process has been demonstrated only recently. However, whether a similar mechanism operates also during embryonic hematopoiesis is unknown. Here we show that TLRs and their adaptor proteins are functionally expressed during early stages of embryogenesis by short-lived maternally-transferred...
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Role of peripheral blood monocytes and innate immunity in diabetes
Zinková, Alžběta ; Daňková, Pavlína (advisor) ; Novota, Peter (referee)
Introduction: Diabetes mellitus is a polygenic disease and its development is influenced to some extent by environmental factors as well. Innate immunity triggers nonspecifically first defense reactions after penetration of the pathogen into the body, while overstimulation components of innate immunity may give rise to autoimmune diseases, including diabetes type 1. The components of innate immunity are, among others, Toll-like receptors (TLRs) belonging to a group of the structures recognizing preserved molecular structures characteristic of pathogens. Toll-like receptors are abundantly expressed by monocytes which produce prolactin (PRL) having an immunostimulatory function. To clarify the role of innate immunity in the pathogenesis of diabetes, we focused on the expression of mRNA and protein expression of TLR2 and TLR4. The expression of PRL was studied only at the level of mRNA. Monocytes were separated by flow cytometry into classical (CD14++) and nonclassical (CD14+). We monitored their percentages and the degree of expression of CD14 antigen on their surface.The operational objective of this dissertation was to optimize the stimulation of monocytes for the planned study of the function of non-pituitary prolactin in vitro and determine the appropriateness of the use of healthy donors' buffy...
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Polymorphism of TLR2/TLR1 and TLR2/TLR6 heterodimers in wild-derived house mouse inbred strains
Bainová, Zuzana ; Vinkler, Michal (advisor) ; Hyršl, Pavel (referee)
Contrary to the classical mouse inbred strains with unnatural genetic variability, wild-derived strains offer a more suitable model for evolutionary immunology. Toll-like receptors (TLRs) belong to initial detectors of invading pathogens. Although TLRs recognise conserved structures they were shown to be polymorphic. This polymorphism is associated with various diseases. In my thesis, I describe variability of Tlr1, 2 and 6 in 24 inbred strains derived from two subspecies of house mouse (Mus m. musculus and M. m. domesticus). These Tlrs exhibit different levels in variability among the strains. In Tlr1 the polymorphic sites are spread along the whole exodomain. Tlr6 is quite conserved (a lower amount of substitutions located far from the binding region and with minor modifications in the amino acid residue properties). Tlr2, on the contrary, contains some substitutions with substantial alternations of residue properties that are located within or nearby the binding region and the subspecies differ at these sites. All alleles of M. m. domesticus and M. m. musculus, except for Tlr1 PWD, Tlr2 STAIL, are phylogenetically separated. The strains and the subspecies vary in the production of IL-1β, IL-12 a NO after stimulation by TLR1, 2 and 6 ligands. This trend is, however, presumably influenced by the effect of...
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Role of toll-like receptors and stress hormone prolactin in defects of immune system
Sluková, Veronika ; Daňková, Pavlína (advisor) ; Hušáková, Markéta (referee)
Introduction: Diabetes mellitus is a polygene disease and on its manifestation have influence also enviromental factors. We have studied the role of extrapituitary prolactin (PRL) and toll-like receptors (TLR) 2 and 4 in the etiopathogenesis of autoimmune diabetes. PRL is mainly produced by hypophysis, but in small concentrations also in the periphery, where it participates in the immune reactions. Therefore, we investigated the influence of the levels of monocytic PRL mRNA on the development of diabetes, and also the influence of G allele of the -1149 G/T polymorphism in the extrapituitary promotor, which has already been associated with other autoimmune diseases. TLRs are receptors of the immune cells that recognize patogenes entering into the body. They play an important role in the iniciation of the immune response. We aimed to find out their function in the pathogenesis of the autoimmune diabetes by the detection of their mRNA levels and protein levels expressed on the cell surface of the monocytes. Material and methods: In this study we included 30 T1D and 21 LADA patients. Three control groups consisted of 23 T2D patients, 23 patients with a nondiabetic disease (neDM) and 60 healthy blood donors (TO). Blood samples have been taken from the individuals. From these blood samples we isolated...
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The innate immunity and circulating monocytes - their significance and function in pathogenesis of coeliac disease.
Němečková, Iva ; Daňková, Pavlína (advisor) ; Palová Jelínková, Lenka (referee)
8 Abstract Introduction: Celiac disease is indentified as the loss of oral tolerance to gluten, it is an organ-specific autoimmune disease in which both, adaptive and innate immunity participate. Monocytes are important part of immune system; they have many functions and express very diverse membrane receptors including Toll-like receptors (TLRs). TLRs are involved in the innate immune response, specifically TLR2 and TLR4 are crucial for recognition of bacterial components and TLR7 recognizes virus's ssRNA. Monocytes also produce prolaktin (PRL), which acts as a cytokine that modulates immune responses. To clarify the role of innate immunity and circulating monocytes in pathogenesis of celiac disease, we focused on changes in expression of selected Toll-like receptors (TLR2, TLR4, TLR7), prolactin, some pro- a anti-inflammatory cytokines (TNF-α, IL-6, IL-12, IL-10). We monitored the influence of the SNP - 1149 G/T on the expression of PRL mRNA. Another objective of this work was the introduction and optimization of in vitro methods for cultivation and stimulation of peripheral monocytes. Material and Methods: This pilot study includes 21 patients with celiac disease and 40 healthy controls. For determination of mRNA levels of the studied genes we isolated RNA from monocytes that were isolated by...
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Modulation of mesenchymal stem cell properties and their use in the regulation of transplantation immunity
Peřinová, Lucie ; Krulová, Magdaléna (advisor) ; Zajícová, Alena (referee)
Mesenchymal stem cells (MSCs) represent a heterogeneous population of stromal cells with a pluripotent differentiation potential. They can be isolated from multiple tissues of mesodermal origin, such as bone marrow, adipose tissue, umbilical cord blood and peripheral blood and afterwards externally expanded according their adherence to the plastic surfaces. These cells show remarkable immunomodulatory properties, suppressing T-, B- and NK-cell functions, and also modulating dendritic cell activities and influencing immune responses during tissue repair and recovery. MSCs have been shown to possess ability to migrate to sites of inflammation and tissue injury. All these properties make MSCs a promising tool for clinical application. Our primary goal was to identify processes that may influence immunoregulatory effects of MSCs. In order to promote immunossupressive qualities of MSCs we established the scheme comprising MSCs precultivated with various cytokines and Toll-like receptors (TLR) ligands in vitro, with the final aim to improve the therapeutic effect of MSCs on wound healing in vivo. We studied modulation of MSCs properties and consequently the effect of influenced MSCs on cells of the immune system. The immunosuppression is mainly mediated through secreted factors that MSCs produced after...
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Physiological role of SIGIRR in early embryonic development.
Hanusová, Zdeňka ; Filipp, Dominik (advisor) ; Brdička, Tomáš (referee)
IL-1 receptor/Toll-like receptor (IL-1R/TLR) supefamily represents a group of proteins that share highly conserved TIR domain in their cytoplasmic region. Signal transduction mediated by TIR-containing proteins involves the activation of NF-κB transcription factor and thus the members of this superfamily play a key role in many physiological responses related to innate immune defense and inflammation. SIGIRR (single immunoglobulin IL1R-related molecule) is a recently discovered member of the IL-1R family, however it differs from the other group members by its unique structural features. SIGIRRhas been so far considered to be an 'orphan' receptor as no SIGIRR ligand has been identified yet. Moreover, SIGIRR itself is not capable to induce the NF-κB activation. Instead, SIGIRR is supposed to act as a negative regulator for IL- 1Rs/TLRs mediated inflammation. Its inhibitory function has been implemented in several signalling pathways in various cell types and tissues including the kidney, the digestive tract and the lung. Recent reports also suggest that SIGIRR could play a role in early embryonic development. The main aim of this thesis is to characterize the mechanism how SIGIRR negative regulatory function in IL-1R/TLR signalling pathway is delivered. Here we describe the establishment of...
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The influence of Toll-like receptors on immunomodulatory properties of mesenchymal stem cells
Heřmánková, Barbora ; Krulová, Magdaléna (advisor) ; Indrová, Marie (referee)
Immunosuppressive properties of mesenchymal stem cells (MSCs) offer potential tools for cell-based regenerative therapy and immunotherapy. MSCs express a large number of Toll like receptors (TLRs) recognizing multiple pathogen-associated molecular patterns (PAMPs), which could be found in patients microenvironment and modulate activity of MSCs. TLR ligands could lead to modulation of proliferation, differentiation, immunosuppression, migration, polarization and production of cytokine. This work discusses the effect of TLRs on immunomodulatory properties of MSCs, because there are lots of contrasts in the results about their effect.
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The effect of tick´s serpin IRS-2 on dendritic cells activated by TLR4 ligand
POSPÍŠILOVÁ, Šárka
IRS-2 is the inhibitor of serine proteases from the Ixodes ricinus tick. My task in this thesis was to find out the effect of the IRS-2 on dendritic cells activated by TLR4 ligand or by Borrelia afzelii. This effect was studied on several levels. I focused on the cytokine production, the expression of costimulatory molecules and cell signaling pathways. The results show that the IRS-2 may inhibit the expression of costimulatory molecules CD-80 a CD-86 on the cell surface, but this finding needs to be confirmed again. The production of cytokines was not affected by the IRS-2. The effect of the IRS-2 on the activity of p38, Erk1/2 nor NF-?B in LPS stimulated cells vas not observed. The fosforylation of STAT 3 in cells activated by the B. afzelii was lowered by the IRS-2.
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