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Vliv rekombinantní bakterie Lactobacillus plantarum na vývoj pylové alergieL
Schwarzer, Martin ; Repa, A. ; Wiedermann, U. ; Hrnčíř, Tomáš ; Daniel, C. ; Pot, B. ; Štěpánková, Renata ; Hudcovic, Tomáš ; Tlaskalová, Helena ; Součková, Martina ; Kozáková, Hana
Germfree mice were monocolonized with recombinant Lactobacillus plantarum producing Bet v 1. Subsequently mice were sensitized by intraperitoneal injections to Bet v 1. We found out that monocolonization with recombinant L. plantarum reduced the level of specific Bet v 1 specific IgE antibody and stimulated production of INF-gamma in spleen cells supernatants. We conclude that monocolonization by recombinant L. plantarum shifts the immune response towards Th-1 direction and that it is a promising vaccine candidate against type I allergy
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Vaccine potentiality of cystatin from tick \kur{Ixodes ricinus}
HARCUBOVÁ, Adéla
Ticks belong to the ectoparasites which are dangerous for the human beings because of the transmission of bacterial, viral, and protozoan pathogens. The development of a vaccine against tick is very important. Cystatins play important role in tick digestion and they have an immunomodulation effects. For this reason cystatins are possible candidates for this vaccines. This thesis focuses on experimental mice vaccination with recombinant cystatin from Ixodes ricinus.
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Characterisation of recombinant cathepsins B of the bird schistosome Trichobilharzia regenti
Dvořáková, Hana ; Mikeš, Libor (advisor) ; Dvořák, Jan (referee)
This study focuses on the recombinant cysteine peptidases - cathepsin B originating in the bird schistosome Trichobilharzia regenti that is unique across the whole family for its ability to migrate through the nerve tissue to the final localization. For invasion, migration, degradation of nutritional proteins and/or evasion of host immune responses, schistosome employs peptidases. This study follows the research done by researchers of Department of parasitology, Faculty of Natural Sciences, Charles University. The main goal of this study was to deepen the characteristics of recombinant cathepsins B originating in T. regenti. In T. regenti, two cysteine peptidases - cathepsins B1 (TrCB1) and B2 (TrCB2) - have been previously characterized. TrCB1 is located in the gut of schistosomula and involved in digestion. TrCB2 occurs in post-acetabular penetration glands of cercariae and probably facilitates penetration. The recombinant pro-cathepsin B (isoforms TrCB1.1, TrCB1.4 and also TrCB2) were expressed in Pichia pastoris yeast system. An attempt was made to produce in P. pastoris the recombinant isoform TrCB1.6, in which the active site cysteine is substituted by glycine. While TrCB2 underwent self-processing in the expression medium, TrCB1.1 and TrC1.4 zymogens were effectively activated only after the...
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Characterization of two members from the multigenic family of one-domain Kunitz-inhibitors from the tick \kur{Ixodes ricinus}
SINGEROVÁ, Barbora
Two new genes encoding proteins Monolaris 1 and Monolaris 2 were isolated from tick Ixodes ricinus. Both cDNA fragments code for 94 aminoacid residues long protein with molecular mass 8,1kDa (Monolaris 1) and 8,3kDa (Monolaris 2). The function of Monolaris 1 was tested by using RNA interference in adult females of Ixodes ricinus that were subsequently fed on guinea-pigs. Body mass, egg mass and mortality were measured to evaluate the effect of gene silencing. Recombinant protein Monolaris 1 was prepared in bacterial expression system and antibodies against this protein were raised by immunization of a rabbit. Antibodies reacted with approximately 190kDa big protein in salivary gland, ovary and gut whereas monomer of Monolaris 1 was not detected in tick saliva, salivary glands and other tissues.
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Advances in the production of antibodies in plants
Angelis, Karel ; Šmídková, Markéta
Plants are the oldest biotechnological production system and as eukaryots have advantage over bacteria to modify expressed proteins. Laboratory model plants like tobacco along with crop plants with established logistics for harvest and storage like potatoes or rice are used for production of desired proteins. Current trend is to replace soil grown plants with liquid cultures of plant cells grown under controlled conditions. In this respect the moss Physcomitrella patens besides ability to grow in liquid culture enables due to efficient homologous recombination to modify posttranslational glycosylation to correspond human. For heterologous expression in plants, recombinant, single-chain molecules composed of only variable VL and VH regions of the original antibody forming binding pocket are constructed and expressed. Recombinant scFv antibodies are then used as a part of therapeutic or diagnostic drugs in oncology.
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