The Diplom's Thesis "Mariology of Louise M. Grignion de Montfort and Leo XIII" The Marian worshiper Louis Maria Grignion of Montfort worked in France as a Catholic priest and missionary at the turn of the 17th and 18th centuries. Grignion, the promoter of the rosary prayer and marian veneration, which Pope Leo XIII blessed in 1888, on the 50th anniversary of his ordination to the priesthood, left in his writings a spiritual message and taching, the elements of which we find in the works of Pope Leo XIII. This Pope is the author of Marian encyclicals promoting the rosary prayer. The tematic similarity in his view of the figure of the Virgin Mary and her influence in history appears in both, Grignion and Leo XIII, in the field of mariological expression and in the inseparable trinitological basis of mariology. The first chapter of this work contains the biography of the priest and missionary Louis M. Grignion of Montfort. It is followed by a biography of Gioacchino Pecci, the pope of the Roman Catholic Church since 1887. The second chapter clarifies the periodic conditionality of Grignion's works and outlines his partial mariology and Marian veneration. The third chapter maps the thinking of Louis M. Grignion implicitly present in the Marian encyclicals of Leo XIII on a general level and then in the...

This thesis was performed within the research project “TrophinOak”, which addresses the impact of multitrophic interactions on the pedunculate oak (Quercus robur) clone DF159. In this frame, the present work focuses on the genetic and physiological mechanisms ruling the interaction of the mycorrhiza helper bacterium (MHB) Streptomyces sp. AcH 505 with microcuttings of DF159 either alone or in presence of the ectomycorrhizal fungus Piloderma croceum or the fungal leaf pathogen oak powdery mildew Microsphaera alphitoides. The work consists of 3 chapters. Chapter 1 characterises the growth of AcH 505 and P. croceum in a soil-based culture system used within the TrophinOak project. Besides the establishment and evaluation of quantification methods of these microorganisms by quantitative real-time PCR, the impact of the soil microbial community and the oak on the bacterium-fungus interaction was investigated, and AcH 505 and P. croceum were visualized by scanning electron microscopy. It was observed that the presence of the soil microorganisms and the oak both affect the bacterium-fungus interaction, and that P. croceum enhances the growth of AcH 505. Chapter 2 presents a study with the oak, AcH 505 and the EM fungus P. croceum, enabling to disentangle the direct effect of the MHB on the oak from the indirect one via the EM symbiosis. The used approach was transcriptomic based on RNA sequencing. It was shown that i) differential gene expression occurred between root and the distant leaf tissues (local vs. systemic effects), different developmental stages and treatments, suggesting that oak specifically coordinates its gene expression patterns, and ii) that genes related to plant growth, defence and DNA modification were dominant among the differential expressed genes, suggesting that these processes play essential roles in both symbiotic interactions investigated. Chapter 3 represents a second transcriptome study, addressing how AcH 505 suppresses powdery mildew infection in oak by analysing RNA Sequencing data from singly- and coinoculated oaks. This study combined the systemic impact of the root associated bacterium with local effects of the leaf pathogen, thereby linking belowground and aboveground interactions. Systemic defence response is induced by the bacterium and further enhanced upon pathogen challenge, suggesting that on the leaf level, some bacterial effectors are recognized as harmful for the plant.

Tinea pedis is a chronic fungal infection of the feet, very often observed in patients who are immuno-suppressed or have diabetes mellitus. The practicing allergist may be called upon to treat this disease for various reasons. Sometimes tinea infection may be mistaken for atopic dermatitis or allergic eczema. In other patients, tinea pedis may complicate allergy and asthma and may contribute to refractory atopic disease. Patients with recurrent cellulitis may be referred to the allergist/immunologist for an immune evaluation and discovered to have tinea pedis as a predisposing factor. From a molecular standpoint, superficial fungal infections may induce a type2 T helper cell response (Th2) that can aggravate atopy. Th2 cytokines may induce eosinophil recruitment and immunoglobulin E (IgE) class switching by B cells, thereby leading to exacerbation of atopic conditions. Three groups of fungal pathogens, referred to as dermatophytes, have been shown to cause tinea pedis: Trychophyton sp, Epidermophyton sp, and Microsporum sp. The disease manifests as a pruritic, erythematous, scaly eruption on the foot and depending on its location, three variants have been described: interdigital type, moccasin type, and vesiculobullous type. Tinea pedis may be associated with recurrent cellulitis, as the fungal pathogens provide a portal for bacterial invasion of subcutaneous tissues. In some cases of refractory asthma, treatment of the associated tinea pedis infection may induce remission in airway disease. Very often, protracted topical and/or oral antifungal agents are required to treat this often frustrating and morbid disease. An evaluation for underlying immuno-suppression or diabetes may be indicated in patients with refractory disease.

Aujourd’hui, plusieurs niveaux de complexité ont émergé dans l’étude des phénotypes T CD4 auxiliaires. 1) le nombre important de cytokines différentes pouvant être secrétées par les lymphocytes T CD4. 2) la multiplicité de signaux pouvant agir durant la différenciation des T CD4 pour spécifier leur profile de sécrétion cytokinique. 3) l’association de ces différents profils de cytokines à des pathologies complexes. Au cours de mon doctorat je me suis concentré sur ces trois niveaux de complexité en étudiant la génération de la diversité cytokinique T CD4 et ses associations aux différents sous types de cancer du sein en utilisant des analyses multivariées et des modèles statistiques. Tout d’abord, j’ai pu construire le premier modèle multivarié de la différentiation T CD4 reliant 37 signaux venant de cellules dendritiques à 18 cytokines T CD4. Utilisant ce modèle pour dériver des prédictions, j’ai pu trouver un nouveau rôle à l’IL-12p70 en tant qu’inducteur de différenciation Th17, mais également comme inducteur spécifique d’IL-17F mais pas d’IL-17A lorsqu’il est combiné à l’IL-1. Ensuite, j’ai étudié l’association de ces cytokines T CD4 avec les différents sous types de cancer du sein connus. J’ai pu trouver que les cytokines Th17 étaient préférentiellement associées avec les cancers du sein dits triple négatifs (TNBC). J’ai pu mettre en évidence qu’une forte signature Th17 était associée à une meilleure survie. De plus, en combinant cette signature Th17 à des scores utilisés pour définir le pronostic clinique, tel que l’index pronostic de Nottingham, j’ai pu proposer une nouvelle et meilleure stratification de la survie de ces patients. / Today several levels of complexity have emerged in the field of T helper cytokines: 1) the important number of distinct cytokines that Th cell can secrete in various combinations; 2) The multiplicity of signals that can act during Th differentiation to define the Th cytokine secretion profiles 3) The associations of these T helper secretion profiles with complex diseases. During my PhD I focused on these three levels of complexity and study the generation of T helper cytokine diversity and its association to breast cancer subtypes using multivariate analysis and statistical modeling. First, I was able to build the first statistical model linking 37 dendritic cell derived signals to 18 T helper cytokines. Using this model to derive in silico predictions, I was able to found a new role for IL-12p70 as a promoter of Th17 differentiation and as a main differential inducer of IL-17F independently of Il-17A in presence of IL-1. Then, studying the associations of the Th cytokine diversity with the different subtypes of human breast cancers, I found that Th17 cytokines were preferentially associated to Triple Negative Breast Cancer (TNBC). I found that TNBC patients with a high Th17 signature had a better survival. In addition, I showed that Th17 can be combined to clinical prognosis assessment scores, such as the Nottingham Prognosis Index, to better stratify TNBC patients in relevant subgroups for survival prognosis assessment.

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<p>  </p> <p>IL-9-producing CD4+ T helper (Th9) cells contribute to inflammatory responses during infection, anti-cancer responses and autoimmune disease. Thus, elucidating the signals that regulate their differentiation is critical for understanding the roles of Th9 cells in protective immunity and disease. Th9 cells differentiate in response to IL-4, TGF-β and IL-2, where IL-2 signaling through STAT5 is crucial for transactivating <em>Il9</em> locus. While the roles of IL-4 and TGF- β-mediated signaling are relatively well understood, how IL-2 signaling contributes to Th9 cell differentiation outside of directly inducing the <em>Il9</em> locus remains less clear. I found that human allergen-induced Th9 cells exhibited a strong signature of STAT5-mediated gene repression that was associated with inhibition of a Th17-like transcriptional signature. Likewise, blockade of IL-2/STAT5 signaling increased IL-17 and RORγt expression in murine Th9 cells <em>in vitro</em>. Interestingly, development of this Th17-like phenotype was independent of STAT3. While STAT3 was not required for IL-17 expression, it was required for their long-term persistence. These results suggest that IL-2/STAT5 signaling controls the balance between Th9 and Th17-like cell differentiation in vitro and during allergy. Additionally, I found that murine Th9 cells cultured in a low IL-2 environment had reduced IL-9 production and a diminished NF-kB-associated transcriptional signature, suggesting that IL-2 signaling is associated with NF-kB activation in Th9 cells. Interestingly, NF-kB activation via IL-1β stimulation enhanced Th9 differentiation under IL-2 limiting conditions and promoted their inflammatory potential in a mouse model of Lung inflammation. Mechanistically, we found that IL-2- limiting conditions enhanced IL-1β receptor expression and that IL-1β/NF-kB signaling increased the sensitivity to IL-2 and silenced the expression of the anti-Th9 transcription factor BCL6. Together, these findings indicate that IL-1β /NF-kB signaling can promote Th9 cell differentiation in IL-2-limiting conditions and that this pathway may be targeted to enhance Th9 differentiation and their inflammatory function.  Collectively, these data revealed two novel roles for the IL-2/STAT5 axis in Th9 cells.</p> <p>The Thymocyte associated High Mobility Group (HMG) box, known as TOX has been previously described to have paramount functions in the development of all the lineages of CD4+ T cells during thymic selection, during CD8+ T cell exhaustion and in Tfh cell differentiation and function. However, the role of TOX in non-Tfh CD4+ T cells in the periphery has not been addressed. In these studies, I found that CD4+ T cells express TOX in the steady state in secondary lymphoid organs like spleen, lymph nodes, and Peyer’s patches. Specifically, TOX was expressed remarkably in Tfh, Th1, Treg cells, and other non-Tfh unidentified Th cells, as well as Th2 cells in the lungs. Transcriptomics analyses using bulk RNA-seq revealed that TOX minimally alters s gene expression, however it revealed for the first time, that TOX induced genes associated with cell migration i.e., <em>Xcl1</em> <em>Ccl3</em>, <em>Ccl4</em> and also the inhibitory cytokine <em>Il10</em>. The induction of IL-10 and CCL3 was validated at the protein levels, and mechanistic studies revealed that the induction of these molecules required the transcription factor BATF, indicating for the first time a mechanism of TOX-mediated functions. Together, these data shed light in novel roles of TOX in CD4+ T cell function and opens the door for future functional and mechanistic studies that may be relevant during health and disease.</p>

Helper T cells (Th) are a vital component of the immune system responsible for directing other immune cells to eliminate pathogens and cancer. Specifically, Th cells facilitate B cell and cytotoxic T cell (Tc) activation and recruitment and enhance their function against cancer and infectious diseases. Th cells are a valuable resource for improving Tc responses in cancer treatment and have become a focus of immunotherapeutic research. While it is increasingly clear that helper T cells serve an important role, the details about which entities produce an effective Th cell response remain unclear. CD5 is a T cell co-receptor that negatively regulates T cell activation and helps fine-tune the TCR repertoire by altering TCR signaling during the selection process in the thymus. This work discusses the role of the co-receptor CD5 in influencing Th cell metabolism, as well as the study of two T cells called LLO118 and LLO56 that have different CD5 expression levels, and their functional response to altered peptides. Antibodies have revolutionized the world of cancer research and accelerated the development of therapies that trigger the immune system to target disease. In recent years, many antibody-based immunotherapies have emerged as effective candidates for combating cancer due to their refined specificity and ability to target a variety of epitopes. However, many therapies, such as those that target CD19 on B cell cancers, are also present on healthy cells, destroying both cancerous and healthy cells alike. Thymidine kinase 1 (TK1) is an enzyme involved in the DNA salvage pathway that converts thymidine into the nucleotide thymine. Recently, TK1 has been shown to be overexpressed on the surface of many cancers such as acute lymphoblastic leukemia. Importantly, TK1 is not expressed on the surface of healthy cells, making it an ideal cancer-specific antigen that can be targeted for cancer treatment. This work discusses our efforts to characterize TK1-specific single-chain antibodies from a yeast display library. According to the World Health Organization, genomics is defined as the study of all genes and their related functions. In contrast to genetics, genomics analyzes the entire DNA makeup of an organism rather than a single gene. In the past 20 years, the cost of genomic sequencing has decreased dramatically, making it affordable and accessible. A key area that must be addressed with genomic testing involves education about their promise, challenges, potential consequences, and ethical considerations. Genomic testing provides a powerful opportunity to educate everyone on scientific and ethical issues to increase understanding on the subject. This work discusses the influence of personal genomics in society and focuses on the importance, benefits, and consequences of genomics education in the classroom, clinic, and the public.

According to the centers for disease control and prevention (CDC) and the world healthorganization (WHO), heart disease and immune related diseases such as diabetes and cancer areamong the leading causes of death around the world. Thus, the regulation of the function ofimmune cell plays a key role in health and disease. Calcium (Ca2+) ions play a critical role inimmune cell activation, function and in a robust immune response. Defects in Ca2+ signalinginfluences the development of cardiac disease, Alzheimer disease, immune cell metabolism,muscle dysfunction, and cancer. Each immune cell is unique in its activation and function,making it relevant to understand how activation of each type of immune cell is regulated. Herewe describe the role of the nBMP2 protein in macrophage activation and function and the role ofthe CD5 co-receptor in helper T cell activation and function.The nuclear bone morphogenetic protein 2 (nBMP2) is the nuclear variant of the bonemorphogenetic protein 2 (BMP2), a growth factor important in heart development, neurogenesis,bone, cartilage and muscle development. To better understand the function of nBMP2, transgenicnBMP2 mutant mice were generated. These mice have a slow muscle relaxation and cognitivedeficit caused in part by abnormal Ca2+ mobilization. Mutant nBMP2 mice also have an impairedsecondary immune response to systemic bacterial challenge. Here we have further characterizedmacrophage activation and function from mutant nBMP2 mice before and after bacterialinfection. We describe how nBMP2 influences the Ca2+ mobilization response and phagocytosisin macrophages, revealing a novel role of the nBMP2 protein in immune cell regulation.CD5 is a surface marker on T cells, thymocytes, and the B1 subset of B cells. CD5 isknown to play an important role during thymic development of T cells. CD5 functions as anegative regulator of T cell receptor (TCR) signaling and fine tunes the TCR signaling response.Here we describe our characterization of CD5 regulation of Ca2+ signaling in naïve helper Tcells. We also outline our findings examining how CD5-induced changes in helper T cellactivation influence other biological processes such as immune cell metabolism, the diversity ofthe gut microbiome, and cognitive function and behavior. Thus, this work elucidates theinfluence of the CD5 co-receptor on the functional outcomes in multiple systems when CD5 isaltered.

T Zellen können in zwei Hauptpopulationen mit unterschiedlichen Aufgaben unterschieden werden. CD4+ T Zellen exprimieren im Zuge ihrer Aktivierung CD40L, welches ein zentraler kostimulatorischer Rezeptor zur Induktion von B-Zell basierter humoraler Immunität, APC Aktivierung und einer effizienten Effektor CD8+ T Zell Entwicklung ist („Helfer-Funktion“). Im Gegensatz dazu sind die zytotoxischen CD8+ T Zellen dazu vorbestimmt, infizierte oder maligne Zellen direkt abzutöten. Jedoch wurde eine Fraktion von CD8+ T Zellen identifiziert, die nach Aktivierung CD40L exprimiert. Bisher ist nicht verstanden, wie in solchen CD8+ T Zellen a) die CD40L Expression reguliert ist, b) wann und wie die Fähigkeit CD40L zu exprimieren implementiert wird und c) was die Folgen für das Immunsystem sind. In dieser Arbeit konnten wir zeigen, dass sowohl in CD4+ als auch in CD8+ T Zellen die CD40L Expression durch DNA-Methylierung regulatorischer Regionen des CD40LG Lokus reguliert wird. Die Demethylierung zentraler Elemente wird im Thymus implementiert, manifestiert sich mit der T-Zell Reifung und geht mit einer zunehmenden Stabilität der CD40L Expression einher. Erhöhte Expression von CD5 und NUR77 in CD40L+ CD8+ SP Thymozyten weisen auf eine positive Selektion mit hoher Affinität gegen Selbst-peptide während der Reifung im Thymus hin, welche das weitere Schicksal der CD40L exprimierenden CD8+ T Zellen beeinflusst. Naive CD40L+ CD8+ T Zellen besitzen ein anderes TCR Repertoire als CD40L- CD8+ T Zellen und reifen im Zuge ihrer Aktivierung bevorzugt zu Gedächtniszellen mit Zytokin- und Chemokinrezeptorprofilen von Tc2, Tc17 und Tc22 Zellen heran. Mit ihrem nicht-zytotoxischen Phänotyp und ihrer Genexpressionsignatur ähneln diese Zellen stark Helfer-CD4+ T Zellen und können von den klassisch zytotoxischen Tc1 und Tc17+1 Zellen durch ihre IL-6R und fehlende SLAMF7 Expression sowie der Expression von Markern die auf eine Fähigkeit in die Haut zu wandern schließen lassen, unterschieden werden. Zusammenfassend zeigen wir hier, dass naive CD8+ T Zellen von den frühsten Entwicklungsstadien im Thymus an nicht homogen sind und die Fähigkeiten über CD40L Expression eine Helferfunktion auszuüben beziehungsweise über die Sekretion zytolytischer Moleküle Zielzellen abzutöten unabhängig vom CD4+ or CD8+ T-Zell Status sind. Zellen mit Zytokin- und Genexpressionsignaturen, die mit denen der CD8+ Helfer-T Zellen übereinstimmen, wurden von uns und anderen in Geweben (Haut, Lunge) identifiziert und tragen zu den verschiedensten autoinflammatorischen Erkrankungen bei. Diese Arbeit insinuiert daher die Notwendigkeit einer grundlegenen Neubewertung der CD8+ T Zell Fähigkeiten und Funktionen in Immunantworten. / The T cell compartment consists of two major subsets with diverse assignments. CD4+ T cells express CD40L upon activation, a central co-stimulatory receptor to induce B cell mediated humoral immunity, activate APCs and prime efficient effector CD8+ T cell development (“helper function”). In contrast, cytotoxic CD8+ T cells are predetermined to kill infected or malignant cells directly. However, a fraction of CD8+ T cells expressing CD40L upon activation was identified. So far, it is not understood in CD8+ T cells a) how CD40L expression is regulated, b) when and how the ability of CD40L expression is implemented and c) what are the implications for the immune system. In this thesis, we found that CD40L expression is regulated by DNA-methylation of regulatory regions of the CD40LG locus in CD4+ as well as CD8+ T cells. The de-methylation of central elements is implemented in the thymus and increases with T cell maturation reflected by enhanced stability of CD40L expression. Elevated CD5 and NUR77 expression of CD40L+ CD8+ SP thymocytes suggests that high affine detection of self-peptides during positive selection in the thymus implements CD40L expression ability and predetermines the fate of the CD40L imprinted CD8+ T cells. CD40L+ naïve CD8+ T cells differ in their TCR repertoire from their CD40L- counterparts and preferentially mature into memory cell subsets with cytokine and chemokine receptor profiles of Tc2, Tc17 and Tc22 cells. With their non-cytotoxic phenotype and gene expression signatures, the CD40L+ memory CD8+ T cell subsets Tc2, Tc17 and Tc22 widely resemble helper CD4+ T cells and can be distinguished from classical cytotoxic Tc1 and Tc17+1 cells by their IL-6R and absent SLAMF7 expression and their skin migratory phenotype. Altogether, we demonstrate that from the earliest developmental stages in thymus onwards naive CD8+ T cells are not homogenous and the abilites to provide “CD40L based help” or “cytotoxicity mediated killing” are independent of the CD4+ or CD8+ T cell status. Cells with helper-type CD8+ T cell cytokine and gene-expression signatures were found at barrier sites (skin, lung) by us and others where they contribute to multiple autoinflammatory diseases. Therefore, this work insinuates the need to revisite CD8+ T cell capablities and function in immune responses.

Interleukin 2 (IL-2) ist ein Zytokin, welches in menschlichen Gedächtnis-T-Helfer-Zellen (Teff Zellen) exprimiert und sekretiert wird und damit die Immunantwort formt. Im Gegensatz dazu wird IL-2 normalerweise nicht in regulatorischen T-Zellen (Treg Zellen) exprimiert, sondern von diesen nur aufgenommen. Durch die Aktivierung des T-Zellrezeptors werden Signalkaskaden induziert, welche zur Aktivierung der Transkriptionsfaktoren AP-1, NFAT, und NF-kB führen. Diese sind entscheidend für die Genexpression von IL-2. Im Rahmen meiner Dissertation habe ich die Regulation der IL-2 Genexpression untersucht. Dabei vergleiche ich die transkriptionelle Regulation in Teff Zellen mit der Regulation in Treg Zellen. Insbesondere konnte ich zeigen, dass die endogene Konzentration der Transkriptionsfaktoren sich auf die Anzahl der IL-2 Produzenten auswirkt, aber nicht auf die Konzentration von IL-2 innerhalb einer Zelle. Deshalb untersuche ich wie sich die Konzentration der Transkriptionsfaktoren auf die Häufigkeit von IL-2 Produzenten auswirkt. Ich nutze die vorhandenen endogenen Konzentrationen und kann damit vorhersagen, dass die Zahl der IL-2 Produzenten entscheidend von der c-fos Konzentration in Teff Zellen abhängt. Mit Hilfe des entwickelten Modells kann ich voraussagen, wie der spezifische Inhibitor U0126 die Häufigkeit von IL-2 Produzenten verringert. Diese Vorhersage wurde durch Experimente belegt. Meine Modelle zeigen weiterhin, dass c-fos und NFATc2 die Häufigkeit der IL-2 Produzenten in Teff Zellen kooperativ regulieren. In Treg-Zellen zeigt meine Analyse, dass alle Transkriptionsfaktoren eine ähnliche sigmoidale Wirkung auf die Häufigkeit der IL-2-Produzenten ausüben. Im Gegensatz zu den Teff Zellen haben alle Transkriptionsfaktor eine ähnliche maximale Wirkung auf Genexpression von IL-2. Mittels eines Inhibitionsmodelles konnte ich zeigen, dass der Treg zellspezifische Transkriptionsfaktor FoxP3 allen aktivierenden Transkriptionsfaktoren entgegenwirkt. / Interleukin 2 (IL-2) is a cytokine expressed in human memory T helper cells (Teff cells) and the secretion of IL-2 shapes the immune response. In contrast, human regulatory T-cells (Treg cells) commonly do not express IL-2. The gene expression of IL-2 is induced by the activation of the T-cell receptor signaling network activating the transcription factors AP-1, NFAT, and NF-kB. These transcription factors are crucial for initiating IL-2 gene expression. Within my thesis I compare the regulation of IL-2 gene expression in Teff cells and Treg cells using experiments and modeling. I demonstrate that the transcription factor concentrations correlate with number of IL-2 producers but do not affect IL-2 concentration per cell. Thus, I investigate how the transcription factor concentration of c-fos, NFATc2, p65, and p-c-jun affects the frequency of IL-2 producing cells as a proxy for the probability of a cell to produce IL-2. Using the endogenous heterogeneity of transcription factor concentrations, I predict that the number of IL-2 producers is critically dependent on the amount of c-fos in Teff cells. I use my model to predict how perturbations of c-fos by the specific inhibitor U0126 decrease the frequency of IL-2 producers in Teff cells. This prediction was than validated by experiments. My models furthermore indicate the cooperative behavior of c-fos and NFATc2 on the level of frequency of IL-2 producers in Teff cells. In Treg cells, I show that all transcription factors exert a similar sigmoidal effect on the frequency of IL-2 producers. In contrast to the effects seen in Teff cells, all transcription factor have a similar maximal effect on the IL-2 gene expression. With an inhibitory model I explore the relation between the Treg cell-specific transcription factor FoxP3 the transcription factors c-fos, NFATc2, p65, and p-c-jun on the frequency of IL-2 producers. This model indicates that FoxP3 counteracts the activating function of NFATc2, AP-1, and also NF-kB.