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Recombinant proteins and synthetic peptides as tools for studies on antibody response against Phlebotomus orientalis
Ferencová, Blanka ; Volf, Petr (advisor) ; Bilej, Martin (referee)
Sand fly saliva contains proteins and peptides that have an important role in bloodfeeding. Some of those proteins are antigenic and repeated sand fly bites result in a specific antibody response of the bitten host. Antigenic salivary proteins of Phlebotomus orientalis, main vector of visceral leishmaniasis in Sudan and Ethiopia, were identified using immunoblot with dog sera. The 5 most promising antigens were expressed in an E. coli bacterial system. Subsequently, these proteins were tested in ELISA with sera of domestic animals from Ethiopia naturally exposed to P. orientalis, and with sera of mice bitten experimentally by this sand fly species. Salivary gland homogenate (SGH) was used as the positive control. The best antigenic properties were detected in two recombinant proteins, Yellow-related protein PorSP24 and ParSP25-like protein PorSP65, especially in tests with sheep and dog sera. However, nonspecific binding of dog sera was also detected using both antigens. In addition, we proved that sera of mice repeatedly bitten by P. papatasi and Sergentomyia schwetzi do not crossreact with SGH and the tested recombinant proteins of P. orientalis. In a second part of this thesis we designed peptides representing epitopes recognized by specific anti-saliva antibodies. Two peptides were derived from...
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Leishmaniasis in domestic and wild animals
Ferencová, Blanka ; Kolářová, Iva (advisor) ; Spitzová, Tatiana (referee)
Leishmaniasis is an infectious disease affecting a wide range of vertebrates, including humans. Leishmania donovani is the causative agent of its most severe form - visceral leishmaniasis. So far animal reservoir has not been proven in the life cycle of this parasite and a man is regarded as the only source of infection. However, zoonotic transmission is demonstrated in the species Leishmania infantum, which together with L. donovani and L. archibaldi belongs into L. donovani complex. A lot of domestic and wild animals living near human dwellings are therefore tested for the presence of Leishmania DNA or anti-Leishmania antibodies, hoping to uncover reservoir host in the life cycle of L. donovani. Sensitivity and specificity of the methods significantly affects the results of undertaken studies. The animals play a significant role in the epidemiology of the disease also as a significant blood source for the vectors. Understanding the ecological interactions between parasite, vector, man, domestic and wild animals could lead to more effective transmission control, thus reducing incidence of leishmaniasis in endemic foci. Key words: Leishmania donovani, visceral leishmaniasis, reservoir, domestic animals, wild animals
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