La sous-unité catalytique p110 de la phosphatidylinositol-4,5-biphosphate 3-kinase (PI3K) est exprimée spécifiquement dans les leucocytes, et possède un rôle central dans la biologie des lymphocytes. Des mutations germinales dominantes gain-de-fonction de PI3KCD (codant p110 ) ont été identifiées récemment chez des patients présentant une immunodéficience combinée caractérisée par une présentation clinique hétérogène incluant des infections récurrentes des voies respiratoires et des lymphoproliférations associées à une virémie EBV et/ou CMV élevée, connue sous le nom de syndromes APDS. L’hétérogénéité des présentations cliniques suggère l’existence de facteurs secondaires, génétiques ou environnementaux. Au cours de ma thèse de doctorat, j’ai rapporté le cas de deux nouveaux patients présentant des anomalies génétiques affectant la voie PI3K. Le premier patient, né dans une famille consanguine, présentait une mutation gain- de-fonction de PIK3CD déjà décrite. Par séquençage de l’exome du patient, nous avons identifié une seconde mutation homozygote non-sens de SEC14L2, un régulateur connu de la voie PI3K. Nous avons analysé le phénotype et la fonctionnalité des lymphocytes du patient, et avons mis en évidence in vitro certains mécanismes de régulation de l’activation lymphocytaire par SEC14L2. Enfin, nous avons réalisé une étude de transcriptome afin d’identifier les voies de régulation dérégulées chez notre patient. Dans l’ensemble, ces résultats ont permis d’identifier le premier co-facteur génétique associé au syndrome APDS. Le second patient est également né dans une famille consanguine et présentait des infections sévères des voies respiratoires et un syndrome d’infection chronique à l’EBV (CAEBV) fatal. Par séquençage de l’exome du patient, nous avons identifié une mutation homozygote rare dans PIK3CD. Une modélisation de la structure de la protéine a montré que l’acide aminé muté se situe dans le domaine catalytique de p110 , dans une boucle phylogénétiquement conservée interagissant avec la sous-unité régulatrice p85↵ de la PI3K, et que la mutation conduisait à la perte de cette interaction. In vitro, la mutation de p110 cause la perte de l’activité PI3K, et les lymphocytes T du patient présentaient une diminution de phosphorylation d’AKT et de p70 S6K, deux cibles de la PI3K. Les lymphocytes T du patient présentaient également une production diminuée d’IFN- et de TNF↵, et un excès de prolifération et de flux calcium suite à la stimulation du TCR. Des lignées cellulaires Jurkat déficientes pour PIK3CD générées par CRISPR/Cas9 ont également présenté un défaut de phosphorylation d’ATK et un excès de prolifération et de flux calcium. L’expression de la version sauvage de p110 dans ces cellules, mais pas de la version mutée, a permis une restauration des réponses normales, prouvant l’implication de la mutation dans les phénotypes cellulaires identifiés. De manière intéressante, nous avons mis en évidence l’existence d’un équilibre entre l’activité PI3K et PLC- 1 lors de l’activation T, qui pourrait être expliqué par une compétition pour l’accès à leur substrat commun, le PIP2. Enfin, nous avons identifié chez notre patient une seconde mutation homozygote délétère dans le gène TNFRSF9. Cette mutation est également retrouvée chez la sœur saine du patient, qui présentait une réplication persistante de l’EBV dans le sang, suggérant que cette mutation pourrait agir comme facteur génétique secondaire. Dans l’ensemble, cette étude à permis l’identification de la première immunodéficience associée à des mutations perte-de-fonction de PIK3CD. / The pathway p110 catalytic subunit of phosphatidylinositol-4,5-biphosphate 3-kinase (PI3K) is selectively expressed in leukocytes and has a central role in lymphocytes biology. Gain-of-function dominant germline mutations of PIK3CD (encoding p110 ) have been recently described in patients presenting a heterogeneous combined immunodeficiency associated with respiratory tract infections, lymphadenopathy and high EBV and/or CMV viremia. The heterogeneous clinical presentation suggests the existence of genetic or environmental modifying factors. In my thesis I report two new patients with different genetic defects causing CID. The first patient, born from a consanguineous family, presented a known gain-of- function mutation of PIK3CD. His clinical presentation was partially compatible with the already described patients. By whole exome sequencing, we identified a homozygous nonsense mutation in SEC14L2, a known regulator of PI3K pathway. We further analyzed the phenotype and functions of patient’s lymphocytes, and performed a transcriptome analysis to better characterize the implication of SEC14L2 mutation in the pathology. The second patient was born from consanguineous family and presented recurrent severe respiratory tract infections and a fatal Chronic Active EBV disease (CAEBV). By Whole Exome Sequencing, we identified a homozygous rare missense mutation in PIK3CD. 3D structure modelization showed that the mutated amino acid is located in p110 catalytic domain, in an evolutionarily conserved loop that interacts with alpha-helix of PI3K regulatory subunit p85a, which interaction is lost in mutated p110 . Phenotyping of patient’s circulating lymphocytes showed increased CD8+ T cells and reduced NK and CD4+ T cells. The mutation in PI3KCD resulted in impaired PI3K activity in vitro and in vivo. Moreover, patient’s T cells exhibited reduced activation-induced phosphorylation of AKT and p70-S6K, two indirect targets of p110 , that we restored by expressing wild type p110 . Patient’s T cells also showed a decreased induction of IFN- and TNF-↵ and an increased proliferation and calcium flux after TCR stimulation. By CRISPR CAS9 technology, we generated Jurkat T- cell lines expressing wild type or mutated PI3K. Jurkat cells expressing mutant PI3K showed decreased AKT phosphorylation and increased calcium flux and proliferation after TCR stimulation, confirming the implication of PIK3CD mutation in patient’s cells phenotype. Interestingly, we highlighted the existence of a balance between PI3K and PLC- 1 activity during T cell activation, that may be due to a competition for access to their shared substrate, the PIP2. Finally, we identified in our patient a second deleterious mutation in TNFRSF9 that is shared by his healthy sister, who also presented persistent EBV replication in blood, suggesting that this additional mutation may act as a modifying genetic factor. Taken together, the results presented in this thesis identified the first loss-of-function mutation in PIK3CD causing CID.

Une réponse immunitaire efficace contre un large spectre de pathogènes ou contre des cellules tumorales nécessite l’activation des lymphocytes T CD4+. L’engagement du récepteur T par un peptide antigénique apprêté sur les produits de classe-II du complexe majeur d’histocompatibilité (peptide-CMH, pCMH) portés par une cellules présentatrices d’antigène (CPAg), conduit à de nombreux remaniements du lymphocyte T. Il s’établit notamment à l’interface entre le lymphocyte T et la CPAg, une structure spécialisée qui est la synapse immunologique (ou synapse immune). La synapse est le siège d’événements de signalisation intenses où diverses molécules de signalisation nécessaires l’amplification et la diversification du signal provenant du TCR sont recrutées. Ces évènements de signalisation sont régulés par l’adaptateur transmembranaire LAT (« Linker for Activation of T cells ») qui est présent à la membrane plasmique ainsi que dans des compartiments intracellulaires. La fraction intracellulaire de LAT est recrutée à la synapse immune et il est proposé que ces compartiments vésiculaires participent à la signalisation lymphocytaire T. L’objectif de ce travail de thèse a été de déterminer les voies de trafic intracellulaire nécessaires au recrutement de la fraction intracellulaire de LAT à la synapse immunologique et de comprendre le rôle de ce transport dans l’activation et la réponse lymphocytaire T. Par des approches d’extinction de l’expression de différentes molécules de transport intracellulaires dans les cellules T Jurkat ou des lymphocytes T CD4+ primaires humains ou par l’utilisation de souris Knock-Out (KO), nous avons mis en évidence plusieurs voies de transport impliquées dans le transport de LAT. Nous avons ainsi mis en évidence que le recrutement de LAT à la synapse immunologique nécessite une voie de sécrétion dépendante de la protéine SNARE vésiculaire VAMP7. L’analyse plus avant du transport de LAT a par ailleurs permis de montrer que LAT est présente dans des compartiments de recyclage alors que VAMP7 est principalement localisée dans l’appareil de Golgi. L’étude de la petite GTPase Rab6 et de la protéine t-SNARE syntaxine-16 qui sont impliquées dans des voies de transport rétrograde entre les endosomes de recyclage et l’appareil de Golgi, a permis de dévoiler que cette voie de transport est requise dans le recrutement de LAT à la synapse immunologique, ainsi qu’à la réponse lymphocytaire T in vitro, ex vivo et in vivo. Enfin, le rôle de la protéine de transport intraflagellaire IFT20, qui a déjà été mis en cause dans le transport du TCR, a été analysé chez la souris et a également montré des défauts de recrutement de LAT à la synapse et dans l’activation lymphocytaire T ex vivo et in vivo. Nos résultats mettent ainsi en évidence que la régulation du transport intracellulaire dans les lymphocytes T joue un rôle crucial dans l’activation lymphocytaire T. Nous proposons ainsi un modèle dans lequel LAT est constitutivement internalisé depuis la membrane plasmique et poursuit une voie de recyclage dépendante de l’appareil de Golgi qui contient la machinerie de sécrétion associé à VAMP7. Cette voie de transport intracellulaire, conditionnerait la resécrétion polarisée de LAT à la synapse immunologique dans les conditions d’activation et une réponse lymphocytaire T robuste. / The immune response against a broad spectrum of pathogens or against tumor cells requires the CD4 + T lymphocytes activation. The triggering of T Cell Receptor (TCR) by peptide-MHC through antigen-presenting cells (APC) leads to numerous T-cell remodeling and the establishment of a specialized structure at the interface between the T lymphocyte and the APC: the immunological synapse. The synapse is the site of intense signaling events where various signaling molecules are recruited in order to amplify and diversify the signal initiated by the TCR. These signaling events are regulated by the transmembrane adapter LAT ("Linker for activation of T cells") which is present at the plasma membrane as well as in intracellular compartments. The intracellular fraction of LAT is recruited at the immune synapse and it is proposed that these vesicular compartments participate in T lymphocyte signaling. The objective of this thesis work was to determine the intracellular trafficking pathways required for the recruitment of the intracellular pool of LAT to the immunological synapse and understand the role of this transport in T cell activation and response. By silencing the expression of different intracellular transport molecules in Jurkat T cells or primary human CD4 + T lymphocytes, or by using Knock-Out (KO) mice, we have highlighted several trafficking pathways involved in the transport of LAT to the immune synapse. We have demonstrated that the recruitment of LAT to TCR activation sites requires a secretion pathway dependent on the vesicular SNARE protein VAMP7. Further analysis of the transport of LAT showed that LAT is present in recycling compartments whereas VAMP7 is mainly located in the Golgi apparatus. The study of the small GTPase Rab6 and the t-SNARE syntaxin-16 protein that are involved in the retrograde transport pathways between the recycling endosomes and the Golgi apparatus, demonstrated that this route of transport is required for the recruitment of LAT to the immunological synapse and for T lymphocyte response in vitro, ex vivo and in vivo as well. Finally, the role of intraflagellar transport protein IFT20, which has already been implicated in the transport of TCR, was analyzed in mice and also showed defects in LAT recruitment to synapse and T lymphocyte activation ex vivo and in vivo. Our results thus show that the regulation of intracellular transport plays a crucial role in T lymphocyte activation. We thus propose a model in which LAT is constitutively internalized from the plasma membrane and pursues a Golgi-dependent recycling pathway that contains the secretion machinery associated with VAMP7. This intracellular transport pathway would thus allow the polarized LAT re-secretion to the immunological synapse under activation conditions and a robust T lymphocyte response.

Em 1993, a Organização Mundial da Saúde declarou a tuberculose (TB) como uma emergência global devido à sua relevância epidemiológica e a necessidade de seu controle. Atualmente, a TB ainda é considerada um problema de saúde pública e requer o desenvolvimento de vacinas e terapias que sejam mais eficazes na sua prevenção e tratamento. Nesse sentido, o Laboratório de Vacinas Gênicas da Faculdade de Medicina de Ribeirão Preto estuda há mais de dez anos a eficácia da vacina gênica DNAhsp65 na profilaxia e terapia da TB. Com o intuito de complementar o conhecimento existente sobre os mecanismos imunes desencadeados pela vacina DNAhsp65, assim como sua associação às drogas convencionais utilizadas no tratamento da TB, objetivou-se neste trabalho a caracterização de células natural killer (NK), T natural killer (NKT), e T ?? na imunoterapia da tuberculose experimental com a vacina DNAhsp65, no tratamento com as drogas rifampicina (RIF) e isoniazida (INH), e na associação DNAhsp65-drogas. Inicialmente, camundongos BALB/c foram infectados com Mycobacterium tuberculosis (Mtb) cepa H37Rv no dia 0 e nos dias 1, 7, 15, 30 e 70 após a infecção, foi promovida a eutanásia dos animais infectados ou não (controle) para análise das células T não convencionais no pulmão por citometria de fluxo. No dia 30 após a infecção, os animais infectados receberam os diferentes tratamentos: vacina DNAhsp65, vetor pVAX1, drogas RIF e INH, ou as drogas em associação à vacina. Dez dias após o fim dos tratamentos, foi promovida a eutanásia dos animais para análise das populações celulares no pulmão e linfonodo por citometria de fluxo, imunohistoquímica e PCR em tempo real. Os animais somente infectados com Mtb apresentaram aumento significativo no número das células NK (CD3-CD49b+), NKT (CD3+CD49b+) e T ?? (CD3+??+) logo na primeira semana após a infecção, e esta diferença em relação aos animais controle permaneceu em até 70 dias após a infecção. Entre as células NK presentes no pulmão, observou-se predominância da subpopulação CD11bhighCD27low em todos os animais. Nos animais infectados, verificou-se aumento significativo das subpopulações de NK: CD11bhighCD27high e CD11blowCD27high, nos dias 7 e 15 e somente no dia 15 após a infecção, respectivamente. Entre a população de células T ?? presentes no pulmão, houve predomínio do fenótipo CD27- em animais controles e infectados nos diferentes tempos experimentais. Quanto aos animais infectados com Mtb e tratados com DNAhsp65, verificou-se aumento significativo de células T ?? produtoras de IFN-? e IL-17 no pulmão, e apesar de não ter sido observada diferença na freqüência de células NK e NKT neste grupo, as células NK apresentavam maior expressão da molécula FasL relacionada à morte celular induzida por apoptose. Nos grupos drogas e DNAhsp65-drogas observou-se aumento da freqüência de células T ?? no pulmão, assim como aumento de células NK produtoras de IL-10 e que expressavam o marcador de ativação CD69. Os resultados deste trabalho mostram mais uma vez a eficácia da vacina DNAhsp65 e da associação DNAhsp65- drogas no tratamento de animais infectados com Mtb e sugerem que células T não convencionais como as células NK, NKT e T ?? podem participar na modulação da resposta immune na TB. Estes achados devem ser levados em consideração no desenho de novas estratégias terapêuticas e também profiláticas para a TB. / In 1993, the World Health Organization declared tuberculosis (TB) as a global emergence due to its epidemical relevance and the need to improve its control. Nowadays, TB still remains a public health problem and requires the development of more effective vaccines and therapies. In this sense, the Laboratory of Genetic Vaccines from the School of Medicine of Ribeirão Preto studies, for more than ten years, the efficiency of the genetic vaccine DNAhsp65 in TB prophylaxis and therapy. In order to complement the knowledge about the immune mechanisms triggered by DNAhsp65 vaccine and by its association with conventional drugs used in TB, our aim in this work was to characterize natural killer (NK), natural killer T (NKT) and gamma-delta (??) T cells in the immunotherapy of experimental tuberculosis with the DNAhsp65 vaccine, in the treatment with rifampicin and isoniazid drugs and in the association DNAhsp65-drugs. Initially, BALB/c mice were infected with Mtb strain H37Rv on day 0, and on days 1, 7, 15, 30 and 70 after infection, infected animals or not (control) were euthanized for lung cell analysis by flow cytometry. On day 30 after infection, infected animals received the following treatment: DNAhsp65 vaccine, pVAX1 vector, rifampicin and isoniazid drugs, or drugs in association with DNAhsp65. Ten days after the end of treatment, animals were euthanized for lung and lymph node cell analysis by flow cytometry, immunohistochemistry and real time PCR. Infected animals showed a significant increase of NK (CD3-CD49b+), NKT (CD3+CD49b+) and ?? (CD3+??+) T cells in the first week of infection and this difference compared to control animals remained until 70 days after infection. Within the lung NK cell population, we observed a predominance of CD11bhighCD27low phenotype in all animals. In infected animals, we verified a significant increase of the following NK cell subpopulations: CD11bhighCD27high and CD11blowCD27high on days 7 and 15, and only on day 15 after infection, respectively. Within the lung ?? T cell population, there was a predominance of CD27- ?? T cell in control and infected animals in the different experimental times. In infected animals and subsequently vaccinated with DNAhsp65, we verified a significant increase in ?? T cells producing IFN-? and IL-17 in the lungs. Although we have not seen any differences in NK and NKT cells in this group, NK cells showed higher expression of FasL molecule related to induced cell death by apoptosis. In DNAhsp65-drugs and drugs groups, we observed an increase in lung ?? T cells frequency, as well as increase in NK cells producing IL-10 and expressing CD69, an activation marker. Our results confirm the effectiveness of DNAhsp65 vaccine and its association with drugs in Mtb infected animals and suggest a modulation in the immune response through unconventional T cells such as NK, NKT and ?? T cells. These findings should be taken into consideration in the design of new therapeutic and prophylactic strategies for TB.

Kung, Wing Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 92-115). / Abstracts in English and Chinese. / Abstract --- p.ii / Acknowledgements --- p.vii / Table of contents --- p.ix / List of Tables --- p.xii / List of Figures --- p.xiii / List of Abbreviations --- p.xv / Chapter Chapter 1. --- General introduction --- p.1 / Chapter Chapter 2. --- Literature review --- p.4 / Chapter 2.1 --- Food allergy and its prevalence --- p.4 / Chapter 2.2 --- Mechanism and clinical symptoms of food allergy --- p.6 / Chapter 2.3 --- Tropomyosin as the major allergen in shellfish --- p.15 / Chapter 2.4 --- Cross reactivity and epitope mapping of tropomyosin --- p.21 / Chapter 2.5 --- Novel approaches for the treatment of food allergy --- p.29 / Chapter Chapter 3. --- Expression of shrimp recombinant tropomyosin and sensitization of mice --- p.36 / Chapter 3.1 --- Introduction --- p.36 / Chapter 3.2 --- Materials and Methods --- p.40 / Chapter 3.2.1 --- "Recovery of E, coli with tropomyosin-carrying plasmid" --- p.40 / Chapter 3.2.2 --- Preparation of tropomyosin-carrying plasmid --- p.41 / Chapter 3.2.3 --- Confirmation of DNA sequence of the tropomyosin --- p.41 / Chapter 3.2.4 --- Identification of the recombinant protein --- p.43 / Chapter 3.2.5 --- Purification of the recombinant protein --- p.43 / Chapter 3.2.6 --- Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.44 / Chapter 3.2.7 --- Concentration measurement of the recombinant tropomyosin --- p.45 / Chapter 3.2.8 --- Mice --- p.46 / Chapter 3.2.9 --- Mice sensitization and challenging --- p.46 / Chapter 3.2.10 --- Tropomyosin-specific IgE level in blood --- p.47 / Chapter 3.2.11 --- Statistical analysis --- p.49 / Chapter 3.3 --- Results --- p.52 / Chapter 3.3.1 --- DNA sequence of the cloned tropomyosin --- p.52 / Chapter 3.3.2 --- Expression and purification of tropomyosin --- p.52 / Chapter 3.3.3 --- Hypersensitivity symptoms after challenge --- p.53 / Chapter 3.3.4 --- Blood tropomyosin-specific IgE level --- p.53 / Chapter 3.4 --- Discussion --- p.62 / Chapter Chapter 4. --- Identification of T cell epitopes --- p.67 / Chapter 4.1 --- Introduction --- p.67 / Chapter 4.2 --- Materials and methods --- p.67 / Chapter 4.2.1 --- Soluble epitope peptide synthesis --- p.68 / Chapter 4.2.2 --- Isolation of spleen cells from mice --- p.69 / Chapter 4.2.3 --- T cell proliferation assay --- p.70 / Chapter 4.3 --- Results --- p.71 / Chapter 4.3.1 --- Splenocyte proliferation to synthetic peptide --- p.72 / Chapter 4.3.2 --- Splenocyte proliferation to synthetic peptides pool --- p.72 / Chapter 4.4 --- Discussion --- p.77 / Chapter Chapter5 --- General conclusion --- p.89 / References --- p.92

Human γδ T cells expressing the Vγ2Vδ2 T cell antigen receptor play important roles in immune responses to microbial pathogens by monitoring prenyl pyrophosphate isoprenoid metabolites. Most adult Vγ2Vδ2 cells are memory cytotoxic cells that produce interferon-γ (IFN-γ). Recently, murine γδ T cells were found to be major sources of interleukin (IL)-17A in anti-microbial and autoimmune responses. To determine if primate γδ T cells play similar roles, we characterized IL-17A and IL-22 production by Vγ2Vδ2 T cells. IL-17A-producing memory Vγ2Vδ2 T cells exist at low but significant frequencies in adult humans (1:2,762 T cells) and at even higher frequencies in adult rhesus macaques. Higher levels of Vγ2Vδ2 T cells produce IL-22 (1:1,864 T cells) although few produce both IL-17A and IL-22. Unlike adult humans where many IL-17A+ V#947;2Vδ2 T cells also produce IFN-#947; (T#947;δ1/17), the majority of adult macaques IL-17A+ Vδ2 T cells (T#947;δ17) do not produce IFN-#947;. To define the cytokine requirements for T#947;δ17 cells, we stimulated human neonatal V#947;2Vδ2 T cells with the bacterial antigen, (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate, and various cytokines and mAbs in vitro. We find that IL-6, IL-1β, and transforming growth factor-β (TGF-β) are required to generate T#947;δ17 cells in neonates whereas T#947;δ1/17 cells additionally required IL-23. In adults, memory T#947;δ1/17 and T#947;δ17 cells required IL-23, IL-1β, and TGF-β but not IL-6. IL-22-producing cells showed similar requirements. Both neonatal and adult IL-17A+ V#947;2Vδ2 T cells expressed elevated levels of retinoid-related orphan receptor-#947;t. Our data suggest that, like Th17 αβ T cells, V#947;2Vδ2 T cells can be polarized into T#947;δ17 and T#947;δ1/17 populations with distinct cytokine requirements for their initial polarization and later maintenance.

Thesis (Ph. D.)--University of Washington, 2000. / Vita. Includes bibliographical references (leaves 106-115).

The peripheral naive CD4 T cell pool is homeostatically regulated through a balance of thymic production, delivery of survival signals and homeostatic proliferation. CD4 recent thymic emigrants (RTEs) have a high T cell receptor excision circle (TREC) content and express high levels of CD31. We report premature thymic involution in RRMS, initiated by reduced numbers of naive CD4 T cells and various naive CD4 T cell subsets in peripheral blood. Further, CXCR4, a receptor involved in emigration from the thymus, and CD127 and Bcl-2 (survival signals) are upregulated in various naive CD4 T cell subsets in RRMS. As a compensatory process, naive CD4 T cells undergo homeostatic proliferation. This proliferation is a form of peripheral positive selection through self-MHC/self-antigen interaction and thus can contribute to the expansion of autoreactive T cells and predispose to development of RRMS.

Equine arteritis virus (EAV) is the causal agent of equine viral arteritis (EVA), a disease of equids. Following EAV infection, up to 70% of stallions may become carriers and continuously shed the virus in their semen for varying time periods. The long-term carrier stallion has an important role in the transmission and maintenance of EAV in horse populations. Recently, it has been demonstrated a correlation between in vitro susceptibility of CD3+ T lymphocytes to EAV infection and establishment of long-term persistent infection among stallions following natural infections. In this study, we investigated whether stallions with in vitro EAV susceptible CD3+ T lymphocytes are at higher risk of becoming long-term carriers compared to those with the resistant phenotype following experimental infection with the KY84 strain of EAV. Furthermore, we investigated whether there is a significant effect of EAV infection on semen quality during acute phase of the infection. The data suggested that the establishment of the long-term carrier state seems to be associated with the in vitro CD3+ T lymphocyte susceptible phenotypes and that reduced semen quality resulted from the combined effect of fever and scrotal edema observed following EAV infection rather than the direct effect of the virus.

Thesis (Ph.D. in Immunology) -- University of Colorado Denver, 2007. / Typescript. Includes bibliographical references (leaves 133-156). Free to UCD affiliates. Online version available via ProQuest Digital Dissertations;

The influence of interferon gamma and specific antibodies on the infection with E. cuniculi in vivo has been studied. Reconstruction of SCID mice with CD4+ T-lymphocytes from BALB/c mice and from mice with defect gene for interferon gamma was used. Effects of the treatment with mouse recombinant interferon gamma and anti-E. cuniculi sera on survival of E. cuniculi infected SCID mice were monitored. The influence of the immunization with E. cuniculi antigen on the survival of E. cuniculi infected mice with defect gene for interferon gamma was examined.