Schistosoma mansoni é um dos agentes causadores da esquistossomose, doença infecciosa negligenciada que afeta milhões de pessoas no mundo. É um platelminto parasitário dióico, com um complexo ciclo de vida, composto de seis estágios. Nos últimos cinco anos, projetos de seqüenciamento em larga escala de etiquetas de genes expressos (ESTs) de Schistosoma geraram uma quantidade razoável de dados que ainda pode ser mais bem explorada. O objetivo central deste trabalho é analisar computacionalmente a expressão gênica de S. mansoni, sob três focos distintos: (i) micro-arranjos de DNA, para os quais descrevemos o desenho e análise de dados de uma plataforma de cDNA (4,600 elementos) e outra de oligonucleotídeos (44,000 elementos). Estão também descritas diversas ferramentas de análise que implementamos e são amplamente usadas em nosso grupo. Os micro-arranjos têm servido de base para vários projetos, como o estudo da resposta do parasita a hormônios e drogas e expressão gênica durante o ciclo de vida. (ii) Identificação in silico de pares de transcritos senso- antisenso com possível ação em trans, potencialmente importantes na regulação gênica. (iii) Análises da coleção de ESTs existentes sob uma perspectiva evolutiva. Através dessa abordagem encontramos genes importantes como um possível inibidor de angiogênese e um regulador da via do mevalonato, conhecido como essencial para a produção de ovos; estes constituem a principal causa de morbidez da esquistossomose. Os resultados aqui apresentados contribuem para o entendimento da complexa biologia inerente ao ciclo de vida de S. mansoni e para acelerar a busca de futuras possibilidades de tratamento. / Schistosoma mansoni is one of the causative agents of schistosomiasis, a neglected infectious disease which affects millions of people worldwide. It is a dioecious parasitic platyhelminth, with a complex life cycle composed of six stages. In the past five years, large scale sequencing projects have generated a reasonable amount of expressed sequence tag (EST) data that can still be better explored. The goal of this thesis is to computationally analyze the S. mansoni gene expression, under three different focuses: (i) DNA microarrays, for which we describe the design and data analyses of a cDNA (4,600 elements) and an oligonucleotide (44,000 elements) platform. We also describe the implementation of several analysis tools which are widely used in our group. Our microarrays are being used in several projects, such as the study of parasite response to drugs and hormones, as well as its gene expression pattern during the life cycle. (ii) In silico identification of possible trans acting natural sense-antisense pairs, potentially important in gene regulation. (iii) Analyses of the available EST dataset under an evolutionary perspective. We have found interesting genes such as a possible angiogenesis inhibitor and a regulator of the mevalonate pathway, known to be essential for egg production; eggs are the main cause of morbidity of schistosomiasis. The results reported here contribute to the understanding of the complex biology underlying the S. mansoni life cycle and to accelerate the search for future possibilities of treatment.

<p>This thesis focuses on the evolutionary interactions between birds and a parasite, the mosquito-borne Sindbis virus (Togaviridae, <i>Alphavirus</i>). In conclusion, the results show that the Sindbis virus is widespread among birds, and that the fitness of infected hosts may be reduced by the virus. Furthermore, viruclearance ability was revealed by male plumage traits, and viraemia was related to hormonal- and social status.</p><p>The distribution of Sindbis virus infections among passerine birds was examined in five areas in Sweden. Almost all species tested were infected, and three species of thrushes weridentified as the main hosts. In a series of experimental infections, greenfinches (<i>Carduelis chloris</i>) kept in aviaries were used ahosts. First, the behavioural consequences of an infection were investigated. During the infection, birds tended to reduce thespontaneous locomotion activity, and when escaping from a simulated predator attack, infected birds had reduced take-off spee Furthermore, when comparing virus clearance rate between male greenfinches, I found that males with large yellow tail ornaments hafaster virus clearance rates as compared to those with smaller ornaments. Thus, male virus clearance ability was honestly revealed by the size of an ornament. Moreover, males with experimentally elevated testosterone levels experienced a delayed, but not increased viraemia as compared to controls. When the relationship between male social ranand viraemia was examined, I found no evidence that high-ranked males suffered reduced rank during the infection. Nevertheless, viraemipatterns of males were related to their social rank, so that low-ranked birds had a delayed viraemia as compared to high-ranked birds. </p>

This thesis focuses on the evolutionary interactions between birds and a parasite, the mosquito-borne Sindbis virus (Togaviridae, Alphavirus). In conclusion, the results show that the Sindbis virus is widespread among birds, and that the fitness of infected hosts may be reduced by the virus. Furthermore, viruclearance ability was revealed by male plumage traits, and viraemia was related to hormonal- and social status. The distribution of Sindbis virus infections among passerine birds was examined in five areas in Sweden. Almost all species tested were infected, and three species of thrushes weridentified as the main hosts. In a series of experimental infections, greenfinches (Carduelis chloris) kept in aviaries were used ahosts. First, the behavioural consequences of an infection were investigated. During the infection, birds tended to reduce thespontaneous locomotion activity, and when escaping from a simulated predator attack, infected birds had reduced take-off spee Furthermore, when comparing virus clearance rate between male greenfinches, I found that males with large yellow tail ornaments hafaster virus clearance rates as compared to those with smaller ornaments. Thus, male virus clearance ability was honestly revealed by the size of an ornament. Moreover, males with experimentally elevated testosterone levels experienced a delayed, but not increased viraemia as compared to controls. When the relationship between male social ranand viraemia was examined, I found no evidence that high-ranked males suffered reduced rank during the infection. Nevertheless, viraemipatterns of males were related to their social rank, so that low-ranked birds had a delayed viraemia as compared to high-ranked birds.

Schistosoma mansoni é um dos agentes causadores da esquistossomose, doença infecciosa negligenciada que afeta milhões de pessoas no mundo. É um platelminto parasitário dióico, com um complexo ciclo de vida, composto de seis estágios. Nos últimos cinco anos, projetos de seqüenciamento em larga escala de etiquetas de genes expressos (ESTs) de Schistosoma geraram uma quantidade razoável de dados que ainda pode ser mais bem explorada. O objetivo central deste trabalho é analisar computacionalmente a expressão gênica de S. mansoni, sob três focos distintos: (i) micro-arranjos de DNA, para os quais descrevemos o desenho e análise de dados de uma plataforma de cDNA (4,600 elementos) e outra de oligonucleotídeos (44,000 elementos). Estão também descritas diversas ferramentas de análise que implementamos e são amplamente usadas em nosso grupo. Os micro-arranjos têm servido de base para vários projetos, como o estudo da resposta do parasita a hormônios e drogas e expressão gênica durante o ciclo de vida. (ii) Identificação in silico de pares de transcritos senso- antisenso com possível ação em trans, potencialmente importantes na regulação gênica. (iii) Análises da coleção de ESTs existentes sob uma perspectiva evolutiva. Através dessa abordagem encontramos genes importantes como um possível inibidor de angiogênese e um regulador da via do mevalonato, conhecido como essencial para a produção de ovos; estes constituem a principal causa de morbidez da esquistossomose. Os resultados aqui apresentados contribuem para o entendimento da complexa biologia inerente ao ciclo de vida de S. mansoni e para acelerar a busca de futuras possibilidades de tratamento. / Schistosoma mansoni is one of the causative agents of schistosomiasis, a neglected infectious disease which affects millions of people worldwide. It is a dioecious parasitic platyhelminth, with a complex life cycle composed of six stages. In the past five years, large scale sequencing projects have generated a reasonable amount of expressed sequence tag (EST) data that can still be better explored. The goal of this thesis is to computationally analyze the S. mansoni gene expression, under three different focuses: (i) DNA microarrays, for which we describe the design and data analyses of a cDNA (4,600 elements) and an oligonucleotide (44,000 elements) platform. We also describe the implementation of several analysis tools which are widely used in our group. Our microarrays are being used in several projects, such as the study of parasite response to drugs and hormones, as well as its gene expression pattern during the life cycle. (ii) In silico identification of possible trans acting natural sense-antisense pairs, potentially important in gene regulation. (iii) Analyses of the available EST dataset under an evolutionary perspective. We have found interesting genes such as a possible angiogenesis inhibitor and a regulator of the mevalonate pathway, known to be essential for egg production; eggs are the main cause of morbidity of schistosomiasis. The results reported here contribute to the understanding of the complex biology underlying the S. mansoni life cycle and to accelerate the search for future possibilities of treatment.

Cuscuta pentagona is an obligate parasitic plant that hinders production of crops throughout the world. Parasitic plants have unique morphological and physiological features, the most prominent being the haustorium, a specialized organ that functions to connect them with their host's vascular system. The Cuscuta haustorium is remarkable in that it enables mRNA movement to occur between hosts and parasite, but little is known about the mechanisms regulating cross-species mRNA transfer or its biological significance to the parasite. These questions were addressed with genomics approaches that used high throughput sequencing to assess the presence of host mRNAs in the parasite as well as parasite mRNAs in the host. For the main experiment Cuscuta was grown on stems of Arabidopsis thaliana and tomato (Solanum lycopersicon) hosts because the completely sequenced genomes of these plants facilitates identification of host and parasite transcripts in mixed mRNA samples. Tissues sequenced included the Cuscuta stem alone, the region of Cuscuta-host attachment, and the host stem adjacent to the attachment site. The sequences generated from each tissue were mapped to host reference genes to distinguish host sequences, and the remaining sequences were used in a de novo assembly of a Cuscuta transcriptome. This analysis revealed that thousands of different Arabidopsis transcripts, representing nearly half of the expressed transcriptome of Arabidopsis, were represented in the attached Cuscuta. RNA movement was also found to be bidirectional, with a substantial proportion of expressed Cuscuta transcripts found in host tissue. The mechanism underlying the exchange remains unknown, as well as the function of mobile RNAs in either the parasite or host. An approach was developed to assay potential translation of host mRNAs by detecting them in the Cuscuta translatome as revealed by sequencing polysomal RNA and ribosome-protected RNA. This work highlights RNA trafficking as a potentially important new form of interaction between hosts and Cuscuta. / Ph. D.

Les parasites exercent une pression de sélection quasiment universelle. Cette thèse aborde les relations hôte-parasite dans le système Lymnaeidae (Gastropoda) Fasciolidae (Trematoda, douves) sous différents aspects, afin de brosser une image de cette interaction et de son évolution. J'ai tout d'abord établi les relations phylogénétiques entre les espèces de Lymnaeidae, puis retracé l'évolution de différents caractères, tels que la susceptibilité à l'infestation par Fasciola hepatica et F. gigantica. Alors que F. hepatica est un parasite généraliste, capable d'infester des mollusques de presque tous les clades de la famille Lymnaeidae, l'infestation par F. gigantica est plus restreinte à un clade. J'ai ensuite étudié plus finement la coévolution entre le parasite F. hepatica et deux de ses hôtes Lymnaeidae (Galba truncatula et Omphiscola glabra) au sein d'une métapopulation, ce qui a confirmé la stratégie généraliste de F. hepatica. En plus, il semblerait que les mollusques parasités et non parasités de G. truncatula aient des différences génétiques, au moins dans cinq des huit populations étudiées. J'ai caractérisé la diversité génétique de deux espèces de mollusques envahissantes, préférentiellement autogames et impliquées dans la transmission de F. hepatica : Pseudosuccinea columella et Lymnaea sp. On trouve une diversité génétique très réduite, chez ces deux espèces, ce qui pourrait faciliter leur expansion géographique et leur infestation par F. hepatica. Ce travail m'a ensuite conduit à mesurer le temps d'attente avant l'autofécondation et la dépression de consanguinité chez ces deux espèces. J'ai trouvé que ces deux espèces sont caractérisées par une dépression de consanguinité très faible et un temps d'attente nul, ce qui confirme les résultats obtenus lors d'une collaboration dans une étude à plus large échelle. Cette thèse souligne l'importance des études en évolution pour comprendre l'épidémiologie des maladies parasitaires. / Parasites constitute a selective pressure to almost all living beings. This thesis addresses the host-parasite interaction in the Lymnaeidae (Gastropoda) Fasciolidae (Trematoda; liver flukes) system through different approaches, with the aim to give a comprehensive image of this interaction and its evolution. First, I established the phylogenetic relationships among Lymnaeidae species, and then mapped the evolution of different characters such as the susceptibility to the infection by Fasciola hepatica and F. gigantica. While F. hepatica is a generalist parasite, capable to infect snails from almost all clades of the Lymnaeidae, infection by F. gigantica is restricted to one clade. Next, I studied the co-evolution between the parasite F. hepatica and two of its intermediate host species (Galba truncatula and Omphiscola glabra) at a finer scale: within a metapopulation. This study confirmed the generalist strategy of F. hepatica. In addition, it seems that parasitized and non-parasitized G. truncatula snails exhibit genetic differences, at least in five out of eight studied populations.I also characterized the genetic diversity of two species of invasive snails involved in the transmission of F. hepatica: Pseudosuccinea columella and Lymnaea sp. We discuss the possible reasons of invasion success in these snails, despite their low genetic diversity, which could facilitate their infection by F. hepatica. Their capacity to respond to parasitism is certainly reduced, all the more that these species are preferential selfers. This work has then led me to measure the waiting time before self-fertilization and inbreeding depression in these two snails. I found that these two species are characterized by low inbreeding depression and present no waiting time, which confirms the results obtained in a collaborative project at larger phylogenetic scale. This thesis strengthens the importance of evolutionary studies to understand the epidemiology of parasitic diseases.

Argulus foliaceus (Linnaeus, 1758) is a member of the branchiuran family Argulidae and has a worldwide distribution, causing major economic impacts for freshwater aquacultured fish species worldwide. In the UK, it has economic impacts for both aquaculture and sports fishing industries. Previous studies observed haemorrhagic and inflammatory responses after Argulus infection, which has been taken to support the idea that the parasite secretes chemicals during the feeding process to assist with the ingestion of blood and epithelial tissue. The present study suggests that the blood-feeding ectoparasite of fish, A. foliaceus, may use similar mechanisms for evading host immune responses to those used by sea lice and other haematophagous arthropods. No previous studies have directly investigated the nature of the bioactive compounds / proteins, assumed to be released from these ectoparasites, and which are considered to contribute to feeding processes and host-parasite interactions during infection. Thus, the work described in this thesis was undertaken with the objective of identifying, describing and characterising the secretory components that have previously been suggested to be secreted from glandular cells associated with the feeding appendages of Argulus foliaceus. The current study applied transcriptomic and proteomic techniques in conjunction with in situ methods to investigate known immunomodulatory genes that may serve a function in parasite-host interactions. Overall, the findings of this project have generated considerable additional knowledge concerning the biology of Argulus spp. and have provided a list of proteins that may be used by the parasite to facilitate feeding processes by secreting these active molecules into the host and hence modulating their immune defence mechanisms. This information can be used as a baseline for developing freshwater lice control strategies to help prevent the spread of Argulosis in aquaculture by applying vaccination as means of control using the candidate antigens described in this study to specifically target Argulus spp. Knowledge generated by the work described in this thesis can also contribute to the development of drugs for controlling Argulus or functional components of feed that may serve to protect fish against this parasite. Furthermore, data from this thesis enhances the knowledge of the distribution of toxin/venom or venom-like substances in crustaceans and arthropods in general.

La mondialisation et les changements globaux actuels ont un impact considérable sur la distribution des espèces et la composition des communautés. Lorsque ces espèces sont impliquées dans une interaction hôte-parasite les changements dans leur répartition peuvent entraîner la (ré)émergence de maladies infectieuses. La fasciolose, maladie causée par les grandes douves du foie (Fasciola hepatica et Fasciola gigantica) est réémergente dans de nombreux points du globe. Il est difficile de mettre en place des programmes de contrôle parce que (1) les hôtes intermédiaires, des mollusques d’eau douce de la famille des Lymnaeidae, sont composés d’un groupe d’espèces cryptiques difficilement identifiables et (2) plusieurs espèces impliquées dans cette maladie sont invasives. L’objectif de cette thèse était d’étudier les facteurs écologiques et évolutifs à large échelle de l’interaction entre la grande douve du foie et ses hôtes intermédiaires susceptibles de favoriser une (ré)émergence de la fasciolose. J’ai, dans un premier temps, développé des approches moléculaires pour reconnaître les espèces cryptiques tant du côté de la grande douve du foie que du côté des Lymnaeidae. L’utilisation d’une des techniques développées sur Galba schirazensis, Galba cubensis et Galba truncatula, trois espèces de limnées, m’a permis d’identifier leur distribution respective et de modéliser leur niches bioclimatiques grâce à l’utilisation de modèles de niches écologiques. Cette approche de modélisation de niches permet d’inférer la distribution potentielle des trois espèces et nous amène à discuter des avantages potentiels de ces modèles dans la gestion de la fasciolose. Dans un second temps, je me suis intéressée à la structuration de la diversité génétique chez des espèces invasives du système grande douve du foie-limnée, par des approches de génétique des populations et de phylogénie. J’ai pu retracer l’histoire de colonisation, les dynamiques démographiques et le système de reproduction chez P. columella, G. schirazensis et G. cubensis. J’ai montré que ces trois espèces font préférentiellement de l’autofécondation entrainant des pertes drastiques de diversité génétiques sur le front d’invasion. G. cubensis présente cependant une coexistence de plusieurs génotypes dans les aires anciennement colonisées. Les différences génétiques entre ces trois espèces sont discutées à la lumière de ce qu’on sait de leur écologie. Enfin ces résultats m’ont permis de discuter de l’avantage d'être autofécondant en cas d’invasion biologique. Pour conclure l’invasion par ces populations d’hôtes intermédiaires dépourvus de diversité génétique pourrait représenter un risque épidémiologique. En effet un parasite a plus de probabilité de circuler dans une population hôte clonale que dans une population polymorphe. Cette thèse fait le lien entre écologie, interactions hôtes-parasites et génétique de l’invasion afin de mieux comprendre les facteurs d’expansion de la fasciolose à échelle globale. / Globalization and the current global change have significant impacts on species distribution and community composition. When these species are involved in a host-parasite interaction, changes in species range distribution can result in the (re)emergence of infectious diseases. Fasciolosis, a disease caused by the liver flukes (Fasciola hepatica and Fasciola gigantica) is reemerging in many parts of the world. It is difficult to implement control programs because (1) the intermediate hosts, freshwater molluscs of the Lymnaeidae family, are composed by a group of cryptic species (2) several species involved in this system are invasive. The objective of this thesis was to study the ecological and evolutionary factors at a large scale in the interaction between the liver fluke and its intermediate hosts that might drive to fasciolosis (re)emergence. First, I developed molecular approaches to quickly identify cryptic species on the two liver flukes and on three Lymnaeidae species. Using one of these techniques, I identify the respective distribution of Galba schirazensis, Galba cubensis and Galba truncatula and infer their respective climatic envelope by ecological niche modelling. We then modelled and projected the potential species distribution ranges. We discussed the contribution of models to predict the species distribution in space and time giving a considerable advantage to control fasciolosis. I then study the genetic diversity structuration in invasive snails involved in the transmission of F. hepatica, using population genetics and phylogeny. I could infer colonization history, population dynamics and reproductive system of Pseudosuccinea columella, G. schirazensis and G. cubensis. I showed that these three species preferentially make inbreeding causing drastic losses of genetic diversity in the invasion front. However G. cubensis presents a coexistence of several genotypes in formerly colonized areas. Genetic differences between these three species are discussed in the light of what we know about their ecology. Overall, these results illustrate how dramatic the reduction in genetic diversity can be for hermaphrodite animals. Finally, we discuss the epidemiological risk related to the invasion by intermediate hosts depleted in genetic diversity. Indeed, a parasite might circulate easily in a clonal host population than in a polymorphic population. In my thesis I linked ecology, host-parasite interactions with genetics of the invasion to better understand the expansion of fasciolosis at a global scale.

Toxoplasma gondii is an obligate intracellular parasite that infects virtually all warm-blooded organisms. Systemic dissemination of T. gondii in the organism can cause life-threatening infection that manifests as Toxoplasma encephalitis in immune-compromised patients. In addition, mounting evidence from epidemiological studies indicates a link between chronic Toxoplasma infection and mental disorders. To better understand the pathogenesis of toxoplasmosis, basic knowledge on the host-parasite interactions and the dissemination mechanisms are essential. Previous findings have established that, upon infection with T. gondii, dendritic cells (DCs) and microglia exhibit enhanced migration, which was termed the hypermigratory phenotype. As a result of this enhanced migration, DCs and microglia are used as vehicle cells for dissemination (‘Trojan horse’) which potentiates dissemination of T. gondii in mice. However, the precise mechanisms behind the hypermigratory phenotype remained unknown. In this thesis, we characterized host-parasite interactions upon infection with T. gondii and investigated the basic mechanisms behind the hypermigratory phenotype of T. gondii-infected DCs and microglia. In paper I, we observed that upon infection with T. gondii, DCs underwent rapid morphological changes such as loss of adhesiveness and podosomes, with integrin redistribution. These rapid morphological changes were linked to hypermotility and were induced by active invasion of T. gondii within minutes. T. gondii-infected DCs exhibited up-regulation of the C-C chemokine receptor CCR7 and chemotaxis towards the CCR7 chemotactic cue, CCL19. In paper II, we developed a 3-dimensional migration assay in a collagen matrix, which allowed us to characterize the hypermigratory phenotype in a more in vivo-like environment. The migration of T. gondii-infected DCs exhibited features consistent with integrin-independent amoeboid type of migration. T. gondii-induced hypermigration of DCs was further potentiated in the presence of CCL19 in a 3D migration assay. In paper III, we identified a parasite effector molecule, a Tg14-3-3 protein derived from parasite secretory organelles. Tg14-3-3 was sufficient to induce the hypermigratory phenotype. Transfection with Tg14-3-3-containing fractions or recombinant Tg14-3-3 protein induced the hypermigratory phenotype in primary DCs and in a microglial cell line. In addition, Tg14-3-3 localized in the parasitophorous vacuolar space and host 14-3-3 proteins were rapidly recruited around the parasitophorous vacuole. In paper IV, we found that mouse DCs dominantly express the L-type voltage-dependent calcium channel, Cav1.3. Cav1.3 was linked to the GABAergic signaling-induced hypermigratory phenotype. Pharmacological inhibition of Cav1.3 and knockdown of Cav1.3 abolished the hypermigratory phenotype in T. gondii infected DCs. Blockade of voltage-dependent calcium channels reduced the dissemination of T. gondii in a mouse model. In paper V, we showed that microglia, resident immune cells in the brain, also exhibited rapid morphological changes and hypermotility upon infection with T. gondii. However, an alternative GABA synthesis pathway was shown to be involved in the hypermigratory phenotype in microglia. In summary, this thesis describes novel host-parasite interactions, including host cell migratory responses and key molecular mechanisms that mediate the hypermigratory phenotype. The findings define a novel motility-related signaling axis in DCs. Thus, T. gondii employs GABAergic non-canonical pathways to hijack host cell migration and facilitate dissemination. We believe that these findings represent a significant step forward towards a better understanding of the pathogenesis of T. gondii infection. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 4: Manuscript. Paper 5: Manuscript.</p>

Das Protozoon Giardia duodenalis ist eine der Hauptursachen für infektiöse Magen-Darm-Erkrankungen. Die zugrundeliegenden Pathomechanismen sind jedoch nach wie vor unklar. Um die Pathogenität G. duodenalis‘ untersuchen zu können, wird ein Modellsystem benötigt, dass die Komplexität des Darmepithels widerspiegelt. Diese Arbeit zeigt die Etablierung eines Zellkultursystems auf der Basis von organoid-abgeleiteten Epithelien unter Verwendung von filter-basierten Zellkultureinsätzen. Wir haben Protokolle für die Etablierung von organoid-basierten Zellkulturen (ODMs) vier verschiedener Wirte zoonotischer Protozoen unter Verwendung eines einzigen Protokolls erstellt. Die Charakterisierung zeigte, dass das Modellsystem erfolgreich die Polarisierung des Darmepithels nachahmt, aus mehreren Zelltypen besteht und eine Infektion ermöglicht. Der Schwerpunkt der Arbeit lag auf der Analyse der durch G. duodenalis induzierten Barrierestörung in ODMs auf Transkriptions-, Protein- und Funktionsebene. Die Infektion von humanen duodenalen Zellen führte zu einem Verlust der epithelialen Barrierefunktion. Mit Hilfe des transepithelialen elektrischen Widerstandes und Dextran Flux wurde eine Erhöhung der Barrieredurchlässigkeit beobachtet. Die Hemmung von zuvor in immortalisierten Zellmodellen beschriebenen Reaktionswegen konnte die Barrierefunktion nicht wiederherstellen. Stattdessen konnten Veränderungen der Ionenhomöostase sowie den Zusammenbruch der zonula occludens nachgewiesen werden. Der beobachtete Phänotyp konnte auf die Aktivierung des cAMP/PKA/CREB-Signalwegs, als einen von mehreren kausalen Faktoren, zurückgeführt werden. Hier zeigen wir die Etablierung eines aus Organoiden abgeleiteten Modells, das die Untersuchung von G. duodenalis Infektionen in vitro ermöglicht. Mit unserem Modell konnten wir eine neue Reihenfolge von Ereignissen entschlüsseln, die einen der Faktoren während symptomatischer Giardiasis darstellt. / The protozoan Giardia duodenalis is a one of the major causes of gastrointestinal illness. Underlying pathomechanisms remain unclear. An in vitro model system that also mimics the complexity of intestinal epithelium is needed to allow pathogenicity studies. This thesis shows the establishment of a cell culture system based on organoid-derived epithelia using permeable cell culture inserts. We have provided guidelines on the establishment of organoid-derived monolayers (ODMs) of four different hosts of zoonotic protozoa using a single protocol. Characterization showed that the model system successfully mimics intestinal polarization, is composed of multiple cell types and allows for infection with multiple protozoan parasites. As the main focus of the thesis, analysis of G. duodenalis-induced barrier breakdown in ODMs was performed on transcriptional, protein and functional level. Infection of human duodenal, organoid-derived monolayers resulted in a time- and dose-dependent breakdown of epithelial barrier function. Barrier permeability increases were observed ranging from ions to macromolecules as measured by transepithelial electrical resistance and Dextran flux. Inhibition of previously proposed key pathogen-induced pathways observed in immortalized cell models did not rescue barrier dysfunction. We could instead show changes in ion homeostasis, and tight junctional breakdown. While none of the previously proposed effector pathways appeared to be responsible, we could pin-point the observed phenotype to activation of the cAMP/PKA/CREB signaling pathway, as one of the factors of the multifactorial barrier breakdown. The establishment of an organoid-derived infection model is shown, allowing the study of in vitro Giardia duodenalis infections. Using this model, we could decipher a new series of events that may be one of the factors causing the intestinal barrier breakdown observed in symptomatic Giardiasis.