Thesis (Ph. D.)--University of Washington, 1999. / Vita. Includes bibliographical references (leaves 112-132).

Thesis (M.Sc.)--Memorial University of Newfoundland, Faculty of Medicine, 1998. / Typescript. Bibliography: leaves 193-209.

Thesis (Ph.D.)--Memorial University of Newfoundland, Faculty of Medicine, 1999. / Includes bibliographical references.

Thesis (Ph. D.)--University of Washington, 2000. / Vita. Includes bibliographical references (leaves 139-193).

T cells are a cornerstone of the mammalian adaptive immune system. A range of T-cell subsets exist that can orchestrate the overall immune response to pathogens or cancers, either by directly killing infected cells or licensing other cells to do so. Dysregulation of this important process can result in immunodeficiency or autoimmunity. Although T cells have been studied extensively over many decades, the detailed mechanisms underlying T-cell activation remain to be fully resolved. This thesis describes the development of new optogenetic approaches for the modulation of T-cell signalling dynamics and the interrogation of key events in T-cell activation to help investigate this question. Optogenetics is a rapidly emerging technique whereby light can be used to control the spatial and temporal activation, or inactivation of signalling pathways at unprecedented resolution. The methods described in this work utilise the blue light-responsive LOV2 photo-domain from the common oat A. Sativa, which is the foundation of the both the ‘LOVTRAP’ and ‘TULIPs’ optogenetic toolkits. T-cell antigen receptor (TCR) microclusters arise early during the interaction between T cells and antigen presenting cells (APCs). These TCR signalling platforms contain the proteins necessary for sustained T-cell activation, yet the processes underlying their formation and dissociation are still not fully characterised as they have been difficult to investigate with current chemical and genetic manipulations of T cells. Using two optogenetics systems combining either LOVTRAP or TULIPs and the microcluster- scaffolding protein LAT (Linker for the Activation of T cells), it was possible to modulate early T-cell signalling events and measure functional outputs in real-time. Unfortunately, the biological limitations of these LAT-based systems meant that they could not be used to quantitatively investigate microcluster formation. However, in an alternative approach, a drug-inducible, light-controllable chimeric antigen receptor was successfully developed that yielded important new insights into the rapid rate of signal decay within the TCR signalling pathway and the temporal dynamics of T-cell activation over several timescales. T cell-dependent bispecific antibodies (TDBs) are a new class of immuno-therapeutics that can specifically direct a T-cell response towards tumours, by crosslinking the TCR complex to a surface- expressed target on the cancerous cells. However, their mechanism of action has not been studied in detail. The close apposition of the T cell and target cells driven by the TDB interaction can result in the steric exclusion of phosphatases, such as CD45, away from the TCR at the TDB-generated cell-cell interface due to their large, rigid extracellular domains. Using the myeloma-expressed antigen, FcRH5, it was found that membrane-proximal epitopes of FcRH5 drive more robust TCR clustering and increased CD45 exclusion than membrane distal epitopes, which strongly correlated with effective killing of the target cell. These findings have important implications for therapeutic design and implementation of TDBs.

Submitted by Johnny Rodrigues (johnnyrodrigues@ufcg.edu.br) on 2018-07-24T16:42:24Z No. of bitstreams: 1 SABRINA ALVES DE FREITAS - DISSERTAÇÃO PPGMAT 2010..pdf: 457483 bytes, checksum: d828740083c1ccca9a0a0f8b45be01d0 (MD5) / Made available in DSpace on 2018-07-24T16:42:24Z (GMT). No. of bitstreams: 1 SABRINA ALVES DE FREITAS - DISSERTAÇÃO PPGMAT 2010..pdf: 457483 bytes, checksum: d828740083c1ccca9a0a0f8b45be01d0 (MD5) Previous issue date: 2010-04 / Capes / Neste trabalho apresentaremos um estudo sobre polinômios centrais ordinários, Z2-graduados e com involução para algumas importantes álgebras na PI-teoria sobre corpos infinitos. Mais precisamente, descreveremos os polinômios centrais Z2-graduados para as álgebras M2(K) (matrizes 2 × 2 sobre um corpo K), M1,1(E) (subálgebra de M2(E) que consite das matrizes cujas entradas da diagonal principal estão em E0 e os da diagonal secundária estão em E1,onde E é a álgebra de Grassmann com unidade de dimensão infinita e E0 e E1 suas componentes homogêneas de graus 0 e 1, respectivamente) e E ⊗ E. Além disso descreveremos os polinômios centrais para E sobre um corpo infinito K de característica diferente de 2 e finalmente os polinômios centrais com involução para M2(K), considerando as involuções transposta e simplética. / In this work we study ordinary, Z2-graded central polinomials and central polinomials with involution for some important algebras in the theory of algebras with polinomial identities, over infinite fields.Namely, we decribe Z2-graded central polinomials for the algebras M2(K) (2 × 2 matrices over a field K), M1,1(E) (subalgebra of M2(E) whose entries on the diagonal belong to E0 and the off-diagonal entries lie in E1, E is the infinite-dimensional unitary Grassmann algebra, E0 is the center of E and E1 is the anticommutative part of E) and E ⊗ E. Also, we describe the central polinomials for e over a field K, with charK ≠ 2 and finally the central polinomial with involution for M2 (K), considering the transpose and the sympletic involutions.

The immune system generates discrete lineages of cells that are designed to respond optimally to environmental cues and infectious agents. Two distinct lineages of T cells, distinguished by expression of either an αβ or γδ T cell receptor (TCR), arise from a common progenitor in the thymus. The type of pathogen and the cytokine milieu directs effector differentiation of αβ T cells in the periphery through the induction of specific transcriptional networks. γδ T cell development is distinct from that of αβ T cells in its ordered rearrangement of TCR genes and the pairing of Vγ and Vδ chains to generate γδ T cell subsets that home to specific tissues. Unlike conventional αβ T cells, γδ T cells express a preactivated or memory phenotype prior to pathogen encounter, and recent evidence indicates that effector functions may be programmed during thymic development. To better understand the development and function of γδ T cells, we analyzed the gene expression profiles of subsets of γδ T cells segregated by TCR repertoire and maturation state in the thymus. We also determined the impact of TCR signaling and trans-conditioning on γδ T cell subset-specific gene signatures by analysis of Itk-/- and Tcrb-/- γδ T cell subsets. Our analysis has defined three stages of γδ T cell subset-specific differentiation, and indicates that γδ T cells may consist of at least two separate lineages, distinguished by the expression of a Vγ2 or Vγ1.1 TCR, that arise from different precursors during thymic development. Key transcriptional networks are established in immature γδ T cells during the first phase of development, independent of TCR signaling and trans-conditioning, with Vγ2+ cells expressing modulators of WNT signaling, and Vγ1.1+ cells expressing high levels of inhibitor of DNA binding 3 (ID3), which regulates E2A/HEB proteins. The second stage involves the further specification of the Vγ2+ subset specific gene signature, which is dependent upon ITK-mediated signals. In the third stage, terminal maturation of γδ T cell subsets occurs, dependent on both TCR and trans-conditioning signals. The expression patterns of Vγ1.1+ subsets that differ in Vδ usage diverge, and all subsets further elaborate and reinforce their effector programming by the distinct expression of chemokine and cytokine receptors. Alteration of WNT signaling or E2A/HEB activity results in subset specific defects in effector programming, indicating that the transcriptional networks established at the immature stage are crucial for the functional maturation of γδ T cells. These data provide a new picture of γδ T cell development, regulated by multiple checkpoints that shape the acquisition of subset-specific molecular signatures and effector functions.

A key step in the development of an adaptive immune response is the activation of naive T cells by dendritic cells (DCs). DCs sample antigens in the periphery and migrate to the lymphoid organs were they provide different signals to T cells: they present antigenic peptides in the context of MHC molecules, express costimulatory or coinhibitory ligands and produce cytokines that influence T cell fate. The integration of these signals will either induce tolerance or lead to the activation and expansion of effector T cells which will mediate the immune response.<p>The costimulatory CD70/CD27 pathway plays important roles in the development of pro-inflammatory Th1 and CTL responses. CD70 expression on DCs has also been described as a molecular switch from tolerance to immunity. Accordingly, its activity is tightly regulated in vivo. The aim of this work was to investigate the mechanisms controlling the expression of CD70 on dendritic cells and CD27 on T lymphocytes.<p>First, we described a cell-extrinsic mechanism of inhibition exerted on DCs by regulatory T cells (Tregs). Indeed, Tregs controlled Th1 priming in vivo and in vitro by downregulating CD70 on DCs. This control involved a transfer of the CD27 receptor to DCs, possibly via the production of CD27-bearing microvesicles by T cells at the immunological synapse. Acquisition of CD27 by DCs induced the internalization of both CD27 and CD70 and probably their lysosomal degradation. As a consequence, DCs were impaired in their ability to efficiently prime Th1 cells. Second, we analyzed CD70 and CD27 expression in the periphery and provided evidence for a cell-intrinsic control of CD27 expression by ectodomain shedding in the gastrointestinal tract.<p>While they efficiently clear infections, inflammatory responses can also be deleterious to the organism. By restraining CD70 expression on DCs, Tregs would promote tolerance and limit inflammation. Interestingly, tolerance is particularly important in the intestines, which are in constant contact with dietary antigens and the commensal microbiota. Accordingly, we propose that a second layer of control of CD27-driven costimulation takes place in the gut :by shedding CD27, T cells would be desensitized for any potential CD70-dependent costimulation.<p>To further investigate the physiological significance of the mechanisms described above, the immune response will be monitored in animals specifically lacking CD27 expression in the Treg population or expressing a nonsheddable CD27 receptor.<p> / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

This dissertation examines a number of issues related to Static and Dynamic Student t Regression Models. The Static Student t Regression Model is derived and transformed to an operational form. The operational form is then examined in a series of Monte Carlo experiments. The model is judged based on its usefulness for estimation and testing and its ability to model the heteroskedastic conditional variance. It is also compared with the traditional Normal Linear Regression Model. Subsequently the analysis is broadened to a dynamic setup. The Student t Autoregressive Model is derived and a number of its operational forms are considered. Three forms are selected for a detailed examination in a series of Monte Carlo experiments. The models’ usefulness for estimation and testing is evaluated, as well as their ability to model the conditional variance. The models are also compared with the traditional Dynamic Linear Regression Model. / Ph. D.

Atlas-Based Segmentation of medical images is an image analysis task which involves labelling a desired anatomy or set of anatomy from images generated by medical imaging modalities. The overall goal of atlas-based segmentation is to assist radiologists in the detection and diagnosis of diseases. By extracting the relevant anatomy from medical images and presenting it in an appropriate view, their work-flow can be optimised. This portfolio-style thesis discusses the research projects carried out in order to evaluate the applicability of atlas-based methods to a variety of medical imaging problems. The thesis describes how atlas-based methods have been applied to heart segmentation, to extract the heart for further cardiac analysis from cardiac CT images, to kidney segmentation, to prepare the kidney for automated perfusion measurements, and to coronary vessel tracking, in order to improve on the quality of tracking algorithms. This thesis demonstrates how state of the art atlas-based segmentation techniques can be applied successfully to a range of clinical problems in different imaging modalities. Each application has been tested using not only standard experimentation principles, but also by clinically-trained personnel to evaluate its efficacy. The success of these methods is such that some of the described applications have since been deployed in commercial products. While exploring these applications, several techniques based on published literature were explored and tailored to suit each individual application. This thesis describes in detail the methods used for each application in turn, recognising the state of the art, and outlines the author's contribution in every application.