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Salivary proteins of sand flies and the immune aspects of Leishmania transmission
Vlková, Michaela ; Volf, Petr (advisor) ; Schwarz, Alexandra (referee) ; Remoue, Franck (referee)
Sand flies serve as the vectors of leishmaniasis and their saliva was shown to affect the outcome of Leishmania infection by immunomodulation of the host. On the other hand, sand fly saliva contains a large scale of farmacologically active proteins that are strongly immunogenous for bitten hosts and specific anti-saliva immunity initiated by repeated sand fly feeding provides protection against Leishmania infection. Specific cell-mediated immunity was shown to be the core of the protectivity; however, our data suggests that the protective immunity has certain limitations. In mice bitten by sand flies for prolonged periods, we observed the desenzitization in term of abrogation of the protective immunity. Thus, we can speculate that the protective effect of immunity is linked solely with the short-term exposure. Nevertheless, our experiments showed that this aspect is also conditioned by the immediate infection after the protective short-term immunization. Taken together, it seems that these limitations may explain the circulation of leishmaniasis in endemic areas, even though humans and animals are frequently immunized by bites of uninfected sand flies. Repeated sand fly feeding on various hosts also promotes production of anti-saliva antibodies that reflect the intensity of exposure. We...
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Salivary proteins of sand flies and the immune aspects of Leishmania transmission
Kindlová, Michaela
Sand flies serve as the vectors of leishmaniasis and their saliva was shown to affect the outcome of Leishmania infection by immunomodulation of the host. On the other hand, sand fly saliva contains a large scale of farmacologically active proteins that are strongly immunogenous for bitten hosts and specific anti-saliva immunity initiated by repeated sand fly feeding provides protection against Leishmania infection. Specific cell-mediated immunity was shown to be the core of the protectivity; however, our data suggests that the protective immunity has certain limitations. In mice bitten by sand flies for prolonged periods, we observed the desenzitization in term of abrogation of the protective immunity. Thus, we can speculate that the protective effect of immunity is linked solely with the short-term exposure. Nevertheless, our experiments showed that this aspect is also conditioned by the immediate infection after the protective short-term immunization. Taken together, it seems that these limitations may explain the circulation of leishmaniasis in endemic areas, even though humans and animals are frequently immunized by bites of uninfected sand flies. Repeated sand fly feeding on various hosts also promotes production of anti-saliva antibodies that reflect the intensity of exposure. We...
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Macrophages in leishmania - sand fly - host interaction
Kratochvílová, Tereza ; Kolářová, Iva (advisor) ; Fialová, Anna (referee)
Sand flies (order Diptera) are vectors of Leishmania parasites (Trypanosomatida), which are inoculated into the host skin together with the vector saliva. Sand fly saliva plays the important role in the Leishmania transmission; in naive host it supresses the host immune response assisting Leishmania to establish the infection, while in repeatedly bitten host it elicits a protective immune response. The submitted thesis focuses on the effect of sand fly saliva on macrophages, the key cells in the infection control. In the first part of the thesis we established a laboratory model L. major - P. papatasi - Balb/c to describe the protective effect of saliva immunization on Leishmania infection development. Immunized mice were protected against Leishmania infection which was reflected in the ear lesion size, parasite load in the ear dermis and draining lymph nodes but also in cytokine production. On the contrary, produced lower amount of nitric oxide, while arginase activity was comparable with nonimmunized group. The IgG antibodies against saliva served as a marker of exposure to sandflies while IgG antibodies against Leishmania antigens served as a marker of infection severity. The experiments were aimed on the possibility of cross-protectivity in Balb/c mice against L. major between closely related...
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Phlebotomine saliva: the perspecives of application of some of their recombinantly produced proteins
Rožníčková, Lucie ; Zídková, Lenka (advisor) ; Štáfková, Jitka (referee)
Sandflies (Diptera: Phlebotominae) are the vectors of one of the most significant tropical diseases, leishmaniosis. Their saliva contains substances with antihemostatic, vasodilatory and immunomodulatory effect. These substances are of great significance in transmission of leishmaniosis, because of their ability to influence the host immune response. Naive host saliva modulate the immune system to such an extent, that it reduces an infective dose and increases the pathogenicity of leishmaniosis. On the other hand, a repeatedly exposed host generates a cellular and antibody immune response, which is able to make the initial development of infection impossible. Repeated exposure of individual sandflies lead to formation of antibodies against components of saliva, which may play a role as a rate exposure indicator and thus the risk of leashmaniosis infection. These proteins and glycoproteins can be produced by recombinant DNA technology. Recombinant vaccine could help in the fight against leishmaniasis. Keywords: saliva, sand fly, leishmania, recombinant proteins.
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Salivary glycoproteins of bloodsucking arthropods
Sumová, Petra ; Volf, Petr (advisor) ; Mikeš, Libor (referee)
During obtaining their blood meal, bloodsucking arthropods salivate into their host. Bloodsucking arthropods' saliva contains wide array of bioactive macromolecules. Host organism develops antibody response against many of these molecules. Due to interspecies variability in salivary protein composition, detection of antibody response may serve as a marker of the exposure to individual species of bloodsucking arthropods. Host antibody response is mostly elicited by proteins or glycoproteins. Glycoproteins contain one or more oligosaccharide chains attached to the protein. Glycoprotein's antigenicity could be caused by either both parts, or by only the protein, or the sugar part. This fact has to be taken into consideration for choice of the expression system for recombinant glycoprotein synthesis. This work summarizes current knowledge about structure, function and features of salivary glycoproteins in various species of bloodsucking arthropods.
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Macrophages and nitric oxide in leishmania - sandfly - host interactions
Kratochvílová, Tereza ; Fialová, Anna (referee) ; Kolářová, Iva (advisor)
Leishmania reside fagolysosome of macrophages immediately after their entry to host where they multiply and consequently infect other macrophages or eventually other cells. A synthesis of a reactive reactant of oxygen and nitrogen is one of the mechanisms that some mammal cells are equipped with and that also contributes to eradication of leishmania. Nitric oxide rising during a metabolic change of L-arginine under the catalysis of NO synthase is of a large importance. Beyond cytotoxic function, nitric oxide is involved in signalling pathways for a neurotransmission (nNOS) and vasorelaxation (eNOS). Not all types of macrophages have ability to produce NO (iNOS). It is a heterogeneous group differing in immunological function and also in physiology. A group of classical activated macrophages represents an effective APC capable of efficient killing of intracellular pathogens. In addition to NO, they also secrete an inflammatory cytokines, which evolve an immune reaction towards to Th1. Contrary to this, a group of alternative activated macrophages is not capable of any efficient antigen presentation and nitric oxide production but produces L-ornithine, which is a precursor of polyamines, which leishmania utilizes for its own intracellular growth. For the mouse model, status of resistance and/or...
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Vliv klíštěcích slin na fagocytózu borelií dendritickými buňkami
MARŠÁLKOVÁ, Eliška
In this study we examined the effect of the tick saliva from I. ricinus and the effect of recombinant protein IRS-2 from the saliva of I. ricinus on dendritic cells derived from the mice bone marrow. We studied their effect on the production of cytokines by dendritic cells after the stimulation by B. burgdorferi, their effect on the expression of genes, that participate in phagocytosis, and the impact of the tick saliva on phagocytosis of B. burgdorferi by dendritic cells.
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Breath Alcohol Tester
Pagáč, Patrik ; Kolář, Radim (referee) ; Chmelař, Milan (advisor)
The theme of bachlor’s project is design of simply breath alkohol tester. A breath alcohol test determines how much alcohol is in your blood by measuring the amount of alcohol in the air you breathe out (exhale).My task is interpret the possibility of measuring the alcohol content in the body. Describe the different methods and compare their accuracy and useability for practical measurements and also used in law enforcement.
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Production and functional characterization of tick salivary protease inhibitors
KOTÁL, Jan
Two cysteine and two serine protease inhibitors from a tick Ixodes ricinus saliva were overexpressed using a prokaryotic overexpression system and refolded to their native state. Both cysteine protease inhibitors were tested as potential antigens for an anti-tick vaccine showing no effect on tick feeding or reproduction. Various immunological methods were employed to test the potential immunomodulatory function of these proteins without success.
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Analyses of glycoproteins from the salivary glands of the tick \kur{Ixodes ricinus}
BUČINSKÁ, Lenka
I characterized several potential glycoproteins in salivary gland extracts from unfed and partially fed females of ticks Ixodes ricinus using enzyme deglycosylation and lectin labeling. Affinity-based (chromatografic) analysis was applied for isolations of glycoproteins with specificity for GNA (mannose), HPA (N-acetylgalactosamine) and MAA II (sialic acid) lectins. GNA specific 120 kDa glycoprotein was isolated from partially fed females and is modified with N-linked glycans containing {$\alpha$}1,3-mannose. Mass spectrometry analyses confirmed the presence carboxypeptidase M in elution fraction gain with GNA affinity chromatography. GNA specific proteins were purified from unfed female salivary gland extracts. MS analyses identified them as proteins similar to arylsulfatase B and cytoskeletal Sojo protein. Proteins (85 and 56 kDa) isolated with HPA affinity chromatography were characterized as Trappin 12, which is a host protein. MAA II lectin was used for labelling and isolation of 100 kDa protein. N-terminal sequence of the MAA II specific protein predicted similarity with a host protein, Siglec 1. Fucose in salivary gland extract was detected with the labelling of AAA, AAL, UEA I and LTL lectins. Results showed that salivary gland extracts contain {$\alpha$}1,2-; {$\alpha$}1,3- and {$\alpha$}1,6- N-linked fucose and O-linked fucose probably as well. GNA specific proteins were detected in partially fed salivary glands acini type II and III using electron transmission microscopy. Fucose was detected on gut and salivary gland structures using fucose-specific lectin AAL.
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