T. gondii interfère avec l'activation des voies de signalisation de NF-kB des cellules hôtes. Ainsi, lors de l’infection par T. gondii, 85% des gènes dépendant de NF-kB sont up-régulés. Un autre facteur de transcription dont l’expression est modulée par le parasite est UHRF1 (Ubiquitin-like,containing PHD and RING finger domains, 1). UHRF1, en se fixant sur le promoteur du gène de la cycline b, induit une répression épigénétique de ce dernier conduisant à un arrêt du cycle cellulaire des cellules infectées en phase G2 et à un arrêt de la prolifération parasitaire. L’analyse in silico du promoteur du gène uhrf1 a montré qu’il possédait 9 sites de fixation de NF-kB. Effectivement nous avons démontré que NF-kB interagit avec le promoteur du gène uhrf1 lors d’une infection par T. gondii. L’expression d’UHRF1 serait donc modulée par NF-kB dans les cellules infectées par T. gondii. Or NF-kB a une régulation différentielle en fonction de la nature de la souche infectante. Là encore, nous avons pu observer une régulation différentielle d’UHRF1 selon la nature de la souche infectante, pouvant être dues à la régulation souche dépendante de NF-kB. La détermination du rôle précis de l’activation d’UHRF1 dans les cellules infectées et l’identification du ou des facteurs parasitaires responsables pourraient permettre de mieux comprendre les mécanismes de persistance intracellulaire du parasite et de découvrir de nouveaux points d’impact thérapeutiques. / T.gondii interferes with the activation of NF-kB signaling pathways. Thus, upon infection by T.gondii, 85% of genes NF-kB-dependent are up-regulated. Another transcription factor whose expression is modulated by the parasite is UHRF1 (Ubiquitin-like, Containing PHD and RINGfinger domains, 1). UHRF1, bind to the gene promoter of cyclin b and induces epigenetic repression of this gene leading to cell cycle arrest in G2 phase of infected cells and stop the proliferation in both infected cells and parasite. In silico analysis of the uhrf1 gene promoter has been shown to possess 9 binding sites of NF-kB. Our study showed that NF-kB actually interacts with the promoter of gene uhrf1 during infection with T. gondii. This suggests that the expression of UHRF1 is modulated by NF-kB in T. gondii-infected cells. In addition we observed differential regulation of UHRF1 depending on the nature of the infecting strain. These variations may also be due to already well-known differential regulation of NF-kB by different strains of T.gondii. Determining the precise role of UHRF1 activation in infected cells and the identification of the parasitic factor responsible of this activation would allow to a better understanding of the mechanisms of intracellular persistence of the parasite and allow to unravel new therapeutic trails.

Toxoplasma gondii é um protozoário pertencente ao filo Apicomplexa, de distribuição mundial, e que infecta diversas espécies hospedeiros, como mamíferos e aves, possuindo como hospedeiros definitivos o gato e outros felídeos, enquanto o homem e outros animais são os seus hospedeiros intermediários. O tratamento é feito, na maioria das vezes, com uma combinação de sulfadiazina e pirimetamina, agindo na via metabólica do ácido fólico, o qual é necessário para a biossíntese de purinas, pirimidinas e certos aminoácidos. Propusemos estabelecer um protocolo de avaliação sobre esse protozoário, assim como padronizarmos uma metodologia por espectroscopia para uma rápida avaliação ou triagem de substâncias potencialmente ativas contra o parasito. Verificamos a atividade das substâncias ácido ursólico e ácido oleanóico, as quais já demonstraram atividade biológica sobre outras espécies de protozoários, como Plasmodium, Trypanosoma cruzi e Leishmania sp. em sistemas in vitro e in vivo. A metodologia colorimétrica pelo MTT, pelo Alamar Blue®, do kit CyQUANT® NF e a contagem manual em meio líquido das formas taquizoítas do parasito em ensaios biológicos in vitro mostram-se inviáveis, pois há grande dificuldade em manter a integridade do parasita em meio de cultura líquido, o qual se mostra sensível à adição de qualquer outro componente que não aqueles necessários para manter sua viabilidade em ambiente extracelular. O ácido ursólico mostrou-se potencialmente ativo in vitro sobre formas intracelulares de T. gondii. Entretanto, o contato de células infectadas com as substâncias avaliadas por um período de 48 horas não resultou em diminuição mais acentuada na porcentagem de células infectadas do que a ocorrida no tratamento de 24 horas. A comparação dos resultados do tratamento pós infecção celular por 24 horas e do pré-tratamento das formas taquizoítas houve diferenças significativas, indicando principalmente maior ação do pré-tratamento sobre T. gondii. Quando administrado na dose de 7 mg/kg/ dia a camundongos infectados, o ácido ursólico não apresentou atividade sobre o parasita. / Toxoplasma gondii is an Apicomplexa protozoan, of worldwide distribution, that infects several species, from mammalians to birds; its definitive hosts are the cats and other felines, while man and other animals are considered as intermediate hosts. Treatment is, generally, a combination of sulfadiazine and pyrimethamine, triggering the metabolic pathway of folic acid, which is necessary for certain purines, pirimidines and aminoacids biosynthesis. We have proposed an evaluation protocol on this protozoan, and also to establish a methodology by spectroscopy for rapid evaluation of pottentialy bioactive substances against the parasite. The ursolic acid and oleanoic acid bioactivity were tested. These substances have already demonstrated to be effective on another protozoan species, like Plasmodium, Trypanosoma cruzi e Leishmania sp., either in vitro or in vivo. The colorimetric methodology by MTT, Alamar Blue®, CyQUANT® NF kit and manual counting of tachyzoite forms in liquid culture medium showed to be unviable, because there is a great difficult to maintain the parasite viability in liquid culture medium, wich one is sensible to addition of any other component different of that necessary for its survival in extracelular ambient. Ursolic acid was pottentialy active on T. gondii intracellular forms. However, the infected cells in contact with the tested substances for a period of 48 hours did not show a statistical greater reduction as compared to infected cells which underwent treatment for 24 hours. Significant results were observed when comparing pre-treatment of tachyzoite forms and treatment for 24 hours post-cellular infection. Our data pointed in the direction that pre-treatment exerted a higher effectiveness. Any parasiticidal activity was observed when ursolic acid on a concentration of 7 mg/kg/day was administered to infected mice.

O trabalho teve por objetivo diagnosticar a infecção experimental e natural por Toxoplasma gondii em pombos (Columba livia), por técnicas sorológicas, biológicas e moleculares. Pombos foram infectados com oocistos esporulados de T. gondii e acompanhados por 60 dias com coleta de soro semanal para o acompanhamento da curva de anticorpos séricos e eutanásia quinzenal para avaliar a presença do parasito em diferentes tecidos. Observou-se concordância em todas as técnicas utilizadas, indicando serem eficazes no diagnóstico da infecção nessa espécie. Pombos de vida livre foram capturados nos municípios de São Paulo, Sorocaba e Ibiúna e anticorpos anti-T. gondii não foram observados. Nestas aves o bioensaio em camundongos foi realizado, independente da ausência de anticorpos e em nenhuma foi possível o isolamento do parasito. / The study aimed to diagnose the experimental and natural infection by Toxoplasma gondii in pigeons (Columba livia), by serological, biological and molecular techniques. Pigeons were infected with sporulated oocysts of T. gondii and monitored for 60 days with weekly serum collection for monitoring the curve of serum antibodies and euthanasia two weeks to assess the presence of parasites in different tissues. Agreement was observed in all the techniques used, indicating to be effective in the diagnosis of infection in this species. Free-living pigeons were captured in the municipalities of São Paulo, Sorocaba and Ibiúna and anti-T. gondii antibodies were not observed. In birds the bioassay was conducted in mice, regardless of the absence of antibodies and none was possible to isolate the parasite.

O presente estudo teve como objetivo avaliar a ocorrência de anticorpos anti-Toxoplasma gondii e anti-Leptospira spp. em peixes-bois (Trichechus inunguis) da Amazônia Brasileira. Amostras sanguineas de 74 peixes-bois foram colhidas. Para a pesquisa de anticorpos anti-T. gondii, os soros foram diluídos a 1:25, 1:50 e 1:500 e a presença de anticorpos foi determinada pela técnica de aglutinação modificada (MAT). Para Leptospira spp. a presença de anticorpos foi determinada pela técnica de soroaglutinação microscópica (SAM), com ponto de corte na diluição de 1:100. Dos 74 peixes-bois 29 (39,2%) foram positivos para T. gondii apresentando título de 25. Para Leptospira spp. 23 (31,1%) foram reagentes para quatro sorovares com títulos variando de 100 a 1600. Os sorovares foram: Patoc, Castellonis, Icterohaemorrhagiae e Butembo. Houve coaglutinação entre os sorovares Patoc, Castellonis e Icterohaemorrhagiae (títulos 100 e 200) em um animal (n°51). Este foi o primeiro relato da ocorrência de T. gondii e Leptospira spp. em peixes-bois da Amazônia brasileira. / This study aimed to evaluate the occurrence of anti-T. gondii and Leptospira spp. in manatees (Trichechus inunguis) from two preservation centers located in the Brazilian Amazon. Blood samples from 74 manatees were taken. For detecting antibodies to T. gondii, sera were diluted 1:25, 1:50 and 1:500 and precense of antibodies was determined by the modified agglutination test (MAT). To Leptospira spp. sera were diluted 1:100 and tested against a collection of 24 antigenic serovars and presence of antibodies was determined by microscopic agglutination test (MAT). Of the 74 manatees 29 (39.2%) were positive to T. gondii with titre at 25. For Leptospira spp. 23 (31.1%) samples were kacted with four serotypes with titers ranging from 100 to 1600. The serovars reagents were Patoc, Castellonis, Icterohaemorrhagiae and Butembo. In one sample, coaglutination of serovars Patoc, Castellonis and Icterohaemorrhagiae (titres 100 and 200) was observed. This is the first report of the occurrence of T. gondii and Leptospira spp. in manatees in the Brazilian Amazon.

A prevalência de Toxoplasma gondii em mamíferos silvestres, quando próximos a áreas urbanizadas, deve ser analisada para diagnosticar riscos à saúde humana e animal. Em Natal-RN, duas Unidades de Conservação (UC), Parque da Cidade e Parque das Dunas, se destacam pela importância ecológica. Nestas UCs a presença de gatos domésticos errantes é constante. O objetivo foi avaliar e comparar a presença de T. gondii através de teste sorológico (MAT) e molecular (gene B1) em gatos e pequenos mamíferos silvestres. Houve associação entre o número de animais silvestres soropositivos em cada parque (p = 0,018), sendo que o Parque da Cidade demonstrou um quadro mais alarmante. A positividade para T. gondii através da amplificação do gene B1 foi positiva em 16,3% do total de animais silvestres analizados e 8,1% do total de gatos avaliados nas duas Ucs. Não houve associação entre o número de animais silvestres positivos para o gene B1 em cada parque (p = 1,00). Existe a participação de ambos os grupos de animais (silvestres e domésticos) no ciclo local do T. gondii. / The prevalence of Toxoplasma gondii in wild mammals close to urbanized areas should be analyzed to diagnose risks to human and animal health. In Natal-RN State, two protected areas, Parque da Cidade and Parque das Dunas are distinguished by ecological importance. In these protected areas, the presence of domestic cats is constant. The aim was to evaluate and compare the presence of T. gondii by serological testing (MAT) and molecular (B1 gene) in cats and small wild mammals. There was an association between the number of wild seropositive animals in each park (p = 0.018). The Parque da Cidade showed a more alarming situation. The positivity for T. gondii by B1 gene amplification was positive in 16.3% on wild animals analyzed and 8.1% on the cats evaluated in the two protected areas. There was no association between the number of wild animals positive for the B1 gene in each park (p = 1,00). There is a participation of both groups of animals (wild and domestic) in the local cycle of T. gondii.

Taquizoítos de T. gondii esterilizados por radiação ionizante é uma vacina interessante para induzir uma imunidade semelhante à infecção, mas sem a formação de cistos. Neste estudo avaliamos as populações celulares do sangue e do baço induzidas pela imunização, a resposta imune humoral, celular e a proteção após desafio com parasitas viáveis. Camundongos foram imunizados com taquizoítos de T. gondii irradiados por v.o. ou i.p.. Os animais foram desafiados com 10 cistos da cepa ME-49 ou VEG por via oral e apresentaram altos níveis de proteção com baixa carga parasitária. Camundongos imunizados por i.p. e v.o. apresentaram anticorpos específicos no soro e o aumento das populações de células B, plasmócitos, células TCD4+ e TCD8+ tanto no sangue como no baço. As células esplênicas de camundongos imunizados por i.p. mostraram a produção de IL-10, IFN-γ e IL-4. Células TCD4+ e células B do baço de camundongos imunizados por i.p. proliferaram após a estimulação com antígeno. A imunização com esse modelo vacinal induziu uma resposta imune mediada com células B, TCD4+ e TCD8+, com aumento da resposta imune humoral e celular que são necessárias para proteção do hospedeiro após uma infecção. Essa resposta imune induzida é uma resposta semelhante a uma infecção natural, sendo assim o desenvolvimento de vacinas utilizando a radiação ionizante como uma ferramenta, pode ser um modelo atrativo e eficiente para testar novos imunógenos no futuro. / Tachyzoites of T. gondii sterilized by ionizing radiation is an interesting vaccine for inducing immunity to infection similarly but without the formation of cysts. In this study we evaluated the cell populations from blood and spleen induced by immunization, the humoral immune response, cellular and protection after challenge with viable parasites. Mice were immunized with irradiated tachyzoites of T. gondii by v.o. or i.p.. The animals were challenged with 10 cysts of the ME-49 or VEG strain orally and showed high levels of protection with low worm burden. Immunized mice by i.p. and v.o. present specific antibodies in the serum and increased populations of B cells, plasma cells, CD4+ and CD8+ T cells in blood and spleen. The spleen cells of immunized mice by i.p. showed the production of IL-10, IFN-γ and IL-4. CD4+ T cells and B cells in the spleen of immunized mice i.p. proliferated upon stimulation with antigen. The immunization with this vaccine model induced an immune response mediated by B cells, CD4+ and CD8+ with increased humoral and cellular immune response are necessary for host protection after infection. This induced immune response is a response similar to natural infection, therefore the development of vaccines using ionizing radiation as a tool, can be an attractive and efficient model for testing new immunogens in the future.

Toxoplasma gondii ist ein weltweit verbreiteter Parasit. In Europa ernährt sich der Endwirt (Katzen) hauptsächlich von kleinen Säugetieren wie Myodes glareolus und Microtus spp. (Wühlmäusen) und Apodemus spp., seltener von Mus spp. Erstere zeigen gegenüber Mus spp. erhöhte Prävalenzen von T. gondii und überleben diese eher als Mus spp. Daher wird vermutet, dass Wühlmäusen und Apodemus spp. eine größere Rolle als Zwischenwirte besitzen, als Mus spp. Der Schutz wird auf das IFN-γ-induzierte IRGb2-b1 zurückgeführt, die den zentralen parasitären Virulenzfaktor ROP5 inhibiert. Daher liegt der Fokus meiner Arbeit auf der protektiven Rolle von IRGb2-b1 bei der Infektion mit T. gondii in Wühlmäusen und Apodemus spp. Mit dieser Arbeit trage ich nützliche Werkzeuge zur Erforschung von Wühlmäusen bei, wie ein rekombinantes M. glareolus IFN-γ Zytokin und neuartige Zellsysteme. Alle untersuchten Wühlmaus-Systeme besaßen einen Phänotyp bei Infektion mit in vivo Resistenz beschrieben wurde: einem IFN-γ-vermittelten Reduktion der Parasitenbürde (mit Wirtszelltod für Typ I Parasiten, ohne für Typ II). Darüber hinaus bestätigen vorläufige Resultate aus Wühlmäusen und Apodemus spp. in Deutschland hohe genetische Vielfalt der IRGb2 Untereinheit zeigen, insbesondere an der vermuteten Grenzfläche zum ROP5, was auf eine Rolle dieses Proteins bei der Infektion hindeutet. Um die Funktion der IRGb2-b1 Proteine zu untersuchen, habe ich ein Zellkultur System, entwickelt, welches erlaubt wild-derived Gene stabil zu exprimieren. Dieses Setup gestattet es, die Auswirkungen von Polymorphismen in Irg Genen bei der Infektion mit T. gondii zu evaluieren. Insgesamt habe ich Werkzeuge erarbeitet, um den Ansatz der Öko-Immunologie in eine Labor-Umwelt zu bringen, wodurch die molekulare Untersuchung ökologisch relevanter Arten möglich wird. Durch Anwendung dieser Werkzeuge stütze ich die Hypothese, dass nicht-Mus Nagetiere, insbesondere M. glareolus, ein bedeutendes Reservoir für T. gondii darstellen. / Toxoplasma gondii is an ubiquitous parasite grouped in three main clonal lineages, type I-III. In Europe, felids, definitive hosts for the parasite, mostly prey on small mammals of Myodes glareolus and Microtus spp. (voles), and Apodemus spp., rather than Mus spp. Voles and Apodemus spp. also display higher T. gondii prevalence and survive infection to a larger extent than Mus spp., although tolerant Mus subspecies exist. This suggests that voles and Apodemus spp. are more relevant intermediate hosts than Mus spp.. Resistance to infection relies on the IFN-γ-induced IRGb2-b1, which inhibits the major parasite virulence factor ROP5. Thus, this work focuses on the protective role of IRGb2-b1 during T. gondii infection in voles and Apodemus spp. With this project I contribute with valuable tools for research on voles, such as the supply of the recombinant M. glareolus IFN-γ cytokine and novel cell systems. All vole systems show a phenotype to type I infection which is associated with in vivo resistance in Mus spp: IFN-γ-mediated host cell death and a decrease in parasite burden. The latter without cell death was observed for type II parasites, suggesting novel protective mechanisms. Further, preliminary results from voles and Apodemus spp. in Germany confirm the expected high diversity in the IRGb2-like subunit, especially at the putative interface with ROP5, which suggests a role of the protein during infection. To assess the role of IRGb2-b1-like in this phenotype, I developed a system which allows establishment of cell lines stably expressing wild-derived Irg-like genes. This setup allows evaluation of the effect of polymorphisms of Irg-like genes during infection. Taken together, I have provided tools to bring eco-immunology into a lab setting to perform molecular investigations of ecologically relevant species. Using these tools, I offer support for the hypothesis that non-Mus rodents, especially M. glareolus, constitutes a relevant T. gondii reservoir in Germany.

Die Toxoplasmose ist eine Infektionskrankheit, die durch den Parasiten Toxoplasma gondii verursacht wird. Während eine Infektion bei den meisten immunkompetenten Menschen asymptomatisch abläuft, kann bei einer Primärinfektion in der Schwangerschaft der Parasit auf den Föten übergehen und zu einem Abort oder zu schweren Erkrankungen dessen führen.Eine pränatale Therapie soll eine Übertragung verhindern oder klinische Auswirkungen beim Kind mindern. Um die Effektivität der in Deutschland durchgeführten Therapie beurteilen zu können, wurde im Rahmen dieser Arbeit eine retrospektive Untersuchung von 685 schwangeren Frauen durchgeführt, deren serologische Befunde auf eine Primärinfektion hindeuteten. In Folge dieser Studie konnte gezeigt werden, dass eine innerhalb von vier Wochen nach Infektion eingeleitete antiparasitäre Therapie das Risiko für die Ausprägung von klinisch-manifesten Erkrankungen des Kindes mindert.Um valide epidemiologische Untersuchungen durchführen zu können ist eine zuverlässige Diagnostik unabdingbar. Hierfür ist von Bedeutung, dass T. gondii ein intrazellulärer Parasit ist, welcher in drei verschiedenen Stadien vorkommt: als Oozysten, Tachy- und Bradyzoiten. Bei einer akuten Infektion sind vorwiegend sich schnell vermehrende Tachyzoiten vorhanden, während bei einer chronischen Infektion Bradyzoiten-enthaltene Zysten dominieren. Es wird davon ausgegangen, dass jedes Parasitenstadium spezifische Antigene exprimiert.Im Zuge dieser Arbeit sollten daher diagnostische Marker identifiziert werden, die eine Differenzierung zwischen einer akuten und einer chronischen Infektion erlauben. Über 2D-Gelelektrophoresen und Massenspektrometrie konnten dabei u. a. die Proteine GRA1, GRA7, ROP9, MIC5 und SUB1 als Marker für eine akute Infektion ermittelt werden. Diese, sowie sieben weitere Proteine, wurden rekombinant in E. coli hergestellt und in einem Lineblot-Verfahren an Humanseren auf ihre diagnostischen Eigenschaften untersucht.Durch den Nachweis spezifischer IgG-Antikörper gegen rGRA1, rGRA2 und rGRA6 konnte eine Toxoplasma-Infektion in bis zu 100 % erkannt werden. Eine Diskriminierung des Infektionsstadiums war allerdings nicht möglich. Eine akute Infektion ließ sich jedoch durch den Nachweis von rSUB1- und rGRA6-spezifischen IgG- und IgA Antikörper mit einer Wahrscheinlichkeit von bis zu 92 % von einer chronischen Infektion unterscheiden.Des Weiteren wurden mit Hilfe des Lineblot-Verfahrens untersucht, ob die rekombinanten Antigene auch Potenzial hatten eine der häufigsten klinisch-manifesten Erkrankungen - die okuläre Toxoplasmose (Retinochorioiditis) - zu identifizieren. Dabei konnten durch den Nachweis GRA2-spezifischer IgA- und BAG1-spezifischer IgG-Antikörper signifikante Unterschiede bei Patienten mit einer Retinochorioiditis im Vergleich zu T. gondii infizierten Patienten ohne okuläre Läsionen festgestellt werden.Um festzustellen, ob der Lineblot-Assay unter der Verwendung rekombinanter Antigene auch in der Veterinärmedizin eingesetzt werden kann, wurden Verlaufseren von Hühnern, Puten und Schweinen untersucht, die experimentell mit Oozysten oder Tachyzoiten infiziert worden waren. Dabei zeigten die Tiere, in Abhängigkeit von dem getesteten Antigen, eine teilweise Infektionsdosis- und Infektionsart-abhängige Antikörper-Kinetik.In der Zusammenschau können die in dieser Arbeit erzielten Ergebnisse zur Verbesserung der Diagnostik und Therapie der Toxoplasmose insbesondere in der Schwangerschaft beitragen.

Effects of Toxoplasmosis on Reaction Times and Prepulse Inhibition of Startle Reaction in Humans vi Abstract Toxoplasma gondii, a single-cell coccidia from almost exclusively parasitic phylum Apicomplexa, does not typically cause acute health issues in humans with most exceptions among immunodeficient individuals and pregnant mothers or, more precisely, their offspring. In the latent phase, the bradyzoites in tissue cysts placed most often in neural and muscle tissues can evolve pressure on the host's body both as a collateral effect of the presence of the parasitic organism in host's tissues and as a consequence of adaptive evolution leading to increase in probability of trophic transmission to the final host, a felid. In humans, this can result in slight changes in personality profiles, deterioration of psychomotor and cognitive functions, and development of serious mental disorders. The thesis focuses predominantly on one of the aspects of the changes, namely the effect of latent toxoplasmosis on the processing of startle signals themselves and when modified by a preceding low-intensity signal; this processing may be connected with the development of schizophrenia in predisposed individuals. Studies conducted within the project framework found changes int the speed of signal processing in...

Die Epithelzellen des Dünndarms bilden die Hauptinfektionsroute für viele Protozoen wie zum Beispiel Toxoplasma gondii und Giardia duodenalis. Jedoch sind die Mechanismen dieser Infektionswege unbekannt, da geeignete Modelle fehlen, welche das Darmepithel nachbilden. In der folgenden Studie wurde eine in-vitro Plattform mit Darmorganoiden (organoid derived monolayers oder ODMs) etabliert, welche man für vergleichende Studien zu Parasit-Wirt-Interaktionen anwenden kann. Das ODM-System wurde angewendet, um die Anfangsphase einer T. gondii-Infektion zu modellieren, wobei der Schwerpunkt auf die Rolle von Interferon gamma (IFNγ) und immunitätsbezogenen GTPasen (Irgs) lag. Es wurde gezeigt, dass sich die Irg-abhängige Kontrolle virulenter Toxoplasma-Stämme zwischen dem Labormausmodell und anderen wildlebenden Nagetierarten unterscheidet. Aus diesem Grund wurden Vergleiche mit Organoiden verschiedener Labormausstämme und der Rötelmaus Myodes glareolus durchgeführt. Myodes glareolus ist eine Nagetierart, von der angenommen wird, dass sie eine höhere Resistenz gegen T. gondii aufweist. Basierend auf die Resultate der quantitativen Immunofluoreszentests und qPCR dieser These, führt die Stimulation mit IFNγ zu einer tendenziell verringerten Replikation der Parasitenstämme RH und Prugniaus in M. glareolus ODMs im Vergleich zu Mus ODMs. In dieser Studie, wurde zum ersten Mal die Rolle von Irgs bei intestinalen T. gondii-Infektionen identifiziert. Zu diesem Zweck wurden Organoide von M. glareolus mit einem fluoreszierend markierten Irgb10-Protein transfiziert, wodurch gezeigt werden konnte, dass Irgb10 T. gondii-Vakuolen dekoriert, was auf eine Beteiligung des Irg-Systems hindeutet. Schließlich wurde ein Koinfektionsmodell für T. gondii und/oder G. Duodenalis in Maus-ODMs etabliert. In diesem Modell wurde gezeigt, dass T. gondii weder die Induktion der Barrierestörung durch G. duodenalis noch die Replikation von T. gondii durch G. duodenalis beeinflusst. / The small intestinal epithelium is the primary route of infection for many protozoan parasites such as Toxoplasma gondii and Giardia duodenalis. Understanding the mechanisms of infection with such parasites, has been hindered due to the lack of appropriate models mimicking the complexity of the intestinal epithelium. Here, an in vitro platform was established, using intestinal organoids (organoid derived monolayers or ODMs) for comparative studies on parasite-host interactions. The ODM system was used to model the events during the early phase of a T. gondii infection, focusing on the role of Interferon gamma (IFNγ) and Immunity Related GTPases (Irgs). Irg dependent control of virulent Toxoplasma strains has been shown to differ between the laboratory mouse model and other wild-derived rodent strains. How these responses occur in rodent species that do not belong to the murine family, is yet to be determined. For this reason, comparisons were made with organoids from different laboratory mice strains and the bank vole Myodes glareolus, a non-muridae rodent species assumed to be more resistant to T. gondii. Based on this thesis, stimulation with IFNγ leads to a trend of reduced replication in M. glareolus ODMs compared to Mus ODMs for both Type I parasite strain RH and for Type II strain Prugniaud, based on quantitative immunofluorescence assays and qPCR. Analysis of the role of Irgs in intestinal T. gondii infections was performed, by transfecting organoids from M. glareolus with a fluorescently labelled Irgb10 protein, showing that Irgb10 decorates T. gondii vacuoles, suggesting Irg-system involvement. Finally, a co-infection model in murine ODMs was established for T. gondii and/or G. Duodenalis. Here, it could be observed that T. gondii did not influence G. duodenalis induction of barrier breakdown nor did G. duodenalis influence T. gondii replication.