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Comparison and characterization of salivary proteins from Sergentomyia and Phlebotomus sand flies
Polanská, Nikola
Sand flies (Diptera, Phlebotominae) are small biting insects and vectors of Leishmania spp. which cause medically and veterinary important disease - leishmaniasis. During the piercing of the host skin, sand fly females inject saliva to facilitate the blood feeding. The sand fly saliva is composed of many bioactive molecules which were shown to possess anti-inflammatory and anti-haemostatic functions. The saliva affects host's immunity in the bite site and consequently enhances the survival and development of transmitted pathogens. Most of the studies focus on salivary proteins and enzymes of sand flies belonging to Phlebotomus and Lutzomyia genera, while salivary proteins from sand flies of the third genus Sergentomyia were neglected so far. In this thesis we focused on comparison of salivary proteins from two Phlebotomus species, namely Phlebotomus perniciosus and Phlebotomus orientalis, and Sergentomyia schwetzi. These sand fly species differ not only by the ecology and geographical distribution but also by host preferences. Both Phlebotomus species prefer large or medium-size mammals as the bloodmeal source, particularly rabbits, hares and dogs for P. perniciosus and cattle, goats, sheep and humans for P. orientalis. Contrarily, Sergentomyia sand flies are known for preferred feeding on reptiles...
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Larval cestodes and tumors: characterization of antibody response and cross-reactivity
Vajs, Vojtěch ; Horák, Petr (advisor) ; Panská, Lucie (referee)
In recent years there have been developments in research concerning the effect of certain species of helminths on cancer. For example, a preventative infection with these parasites is capable of slowing or completely preventing the development of certain cancers in experimental models. In our preliminary experiments, the suppressive effect of the infection with Taenia crassiceps and Mesocestoides corti on melanoma cancer cells was discovered. Primary infection with these tapeworms has been able to suppress and completely prevent the growth and metastasis of the B16F10 melanoma cell line. Parasites affect the immune system of their hosts with their presence and products. It was therefore the subject of this work to explore one of the ways how these tapeworms may achieve this effect - cross-reactivity of antibodies with tapeworm and cancer cell antigens. First the antibody response of the mouse organism to the tapeworm infection was examined in C57BL/6J inbred and ICR outbred strains. The growth of immunoglobulin levels was measured with ELISA within the first five weeks post infection, upon which the antigens, with which these immunoglobulins react, were specified with the help of Western blot. After this, these reacting antigens were localised in the tapeworm organism with immunohistochemical...
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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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Characterization and antigenic properties of salivary yellow-related proteins in phlebotomine sand flies
Cikrtová, Petra
2 ABSTRACT Yellow-related proteins (YRPs) form an abundant protein family, whose members were found in salivary gland transcriptomes of all sand fly species studied up to date. This protein family belongs to the group of MRJP/Yellow proteins occurring in insects and some other organisms. Though sharing similar folding as a six-bladed β-propeller comprising central tunnel, MRJP/Yellow proteins adopted different functions. The structure and biogenic amine-binding property described for the sand fly salivary YRPs was based on a single study conducted on Lutzomyia longipalpis. In the present work, we have modelled the structures of 32 salivary YRPs belonging to 13 different sand fly species. We have shown the general structural similarity of these proteins along with both inter- and intra-specific differences in surface charge, tunnel parameters and in amino acids composition of the amine-binding motif. These modifications indicated divergence in function of individual YRPs, which was experimentally verified in the second project focused on identification of the amine-binding properties of YRPs in two important vectors of Leishmania; Phlebotomus perniciosus and P. orientalis. In each species, two YRPs differ in affinities for biogenic amines serotonin, histamine and catecholamines. However, in both sand fly...
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Comparison and characterization of salivary proteins from Sergentomyia and Phlebotomus sand flies
Polanská, Nikola ; Volf, Petr (advisor) ; Martin-Martin, Ines (referee) ; Chmelař, Jindřich (referee)
Sand flies (Diptera, Phlebotominae) are small biting insects and vectors of Leishmania spp. which cause medically and veterinary important disease - leishmaniasis. During the piercing of the host skin, sand fly females inject saliva to facilitate the blood feeding. The sand fly saliva is composed of many bioactive molecules which were shown to possess anti-inflammatory and anti-haemostatic functions. The saliva affects host's immunity in the bite site and consequently enhances the survival and development of transmitted pathogens. Most of the studies focus on salivary proteins and enzymes of sand flies belonging to Phlebotomus and Lutzomyia genera, while salivary proteins from sand flies of the third genus Sergentomyia were neglected so far. In this thesis we focused on comparison of salivary proteins from two Phlebotomus species, namely Phlebotomus perniciosus and Phlebotomus orientalis, and Sergentomyia schwetzi. These sand fly species differ not only by the ecology and geographical distribution but also by host preferences. Both Phlebotomus species prefer large or medium-size mammals as the bloodmeal source, particularly rabbits, hares and dogs for P. perniciosus and cattle, goats, sheep and humans for P. orientalis. Contrarily, Sergentomyia sand flies are known for preferred feeding on reptiles...
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Characterization and antigenic properties of salivary yellow-related proteins in phlebotomine sand flies
Cikrtová, Petra
2 ABSTRACT Yellow-related proteins (YRPs) form an abundant protein family, whose members were found in salivary gland transcriptomes of all sand fly species studied up to date. This protein family belongs to the group of MRJP/Yellow proteins occurring in insects and some other organisms. Though sharing similar folding as a six-bladed β-propeller comprising central tunnel, MRJP/Yellow proteins adopted different functions. The structure and biogenic amine-binding property described for the sand fly salivary YRPs was based on a single study conducted on Lutzomyia longipalpis. In the present work, we have modelled the structures of 32 salivary YRPs belonging to 13 different sand fly species. We have shown the general structural similarity of these proteins along with both inter- and intra-specific differences in surface charge, tunnel parameters and in amino acids composition of the amine-binding motif. These modifications indicated divergence in function of individual YRPs, which was experimentally verified in the second project focused on identification of the amine-binding properties of YRPs in two important vectors of Leishmania; Phlebotomus perniciosus and P. orientalis. In each species, two YRPs differ in affinities for biogenic amines serotonin, histamine and catecholamines. However, in both sand fly...
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Development of vaccines against porcine circovirus type 2
Janovec, Václav ; Drda Morávková, Alena (advisor) ; Pokorná, Dana (referee)
Porcine circovirus type 2 is a single stranded DNA virus that belongs to the genus Circovirus in the family Circoviridae. This virus is associated with many kinds of diseases in pigs and causes significant economic losses in swine-breeding. In this study, two approaches of vaccination were tested in order to develop an effective vaccine against PCV2. The first approach was to test DNA vaccines. For this purpose, eukaryotic expression plasmids encoding two form of PCV2 Cap protein were constructed. The expression plasmids encoding murine TNF-α and IFN-α1 were also prepared for co-immunization with antigen encoding plasmid to enhance the immune response. The second approach is based on the previous finding that chimeric pentamers of VP1 mouse polyomavirus capsid protein fused with PCV2 can induce protective immunity against PCV2. These chimeric pentamers were further modified by AA substitutions in PCV2 Cap immunodominant epitope in order to enhance protective antibody response directed against PCV2. The chimeric pentamers and DNA vaccines were tested for ability to induce antibody immune response against PCV2 in mice. The results showed that chimeric pentamers are more potent inducers of protective antibody immune response against PCV2 compared to DNA vaccines. However, the protective antibody...
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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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