Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. institutet, 2006. / Härtill 5 uppsatser.

Thesis (Ph.D.)--University of Alabama at Birmingham, 2009. / Title from PDF title page (viewed on Sept., 2009). Includes bibliographical references.

Thesis (Ph.D.)--University of Alabama at Birmingham, 2007. / Title from PDF title page (viewed on Sept. 16, 2009). Includes bibliographical references.

Thesis (Ph.D.)--University of Alabama at Birmingham, 2007. / Title from PDF title page (viewed on Sept. 16, 2009). Includes bibliographical references.

Thesis (Ph. D.)--University of Washington, 2005. / Vita. Includes bibliographical references (leaves 92-100).

Bien que la description qualitative et phénoménologique de la migration cellulaire soient décrites de façon minutieuse dans son approche holistique, des mécanismes de base et connus depuis longtemps n’ont pas encore été explorés quantitativement avec une approche "bottom-up". Un de ces exemples est le comportement migratoire en deux dimensions des lym- phocytes T effecteurs humains. Alors que leur comportement in vivo est déjà connu depuis longtemps et décrit qualitativement, une approche quantitative in vitro offre de nombreuses perspectives. Les interactions des intégrines LFA-1 et VLA-4 avec leurs ligands respectifs ICAM-1 et VCAM-1 ont déjà été étudiées et sont les principales molécules impliquées dans la migration des lymphocytes T effecteurs. Du fait de leur importance dans l’organisation de la mobilité, ces deux protéines sont les principaux objets d’étude de cette thèse. La majorité des travaux précédents ont été réalisés en observant les lymphocytes T dans des tissus vi- vants dont la composition et la densité des molécules d’adhésion ne peuvent ni être déterminées ni contrôlées de façon précise. Nous avons developpés des substrats artifi- ciels permettant d’imiter et de contrôler ces caractéristiques adhésives afin d’examiner et de relier les propriétés physiques des cellules tels que la vitesse ou l’index de mi- gration orientée, avec une réponse cellulaire donnée ainsi que d’associer un type de mouvement avec des interactions intégrines-ligands spécifiques. Pour aller plus loin, nous avons à nouveau analysé la conformation des intégrines puis l’avons modulé pour altérer leur affinité et changer les propriétés précédemment décrites. / The ability of T-lymphocytes to migrate to sites of inflammation towards all different types of tissues based on the interplay between biochemical and mechanical signaling is unique among human cells and underlines the importance of their complex motility apparatus relying on multiple stimuli. A crucial part within the leukocyte adhesion cascade is the firm attachment of the immune cell to the inner wall of the blood vessel and the subsequent migration along its surface until crossing the endothelial cell barrier. These migrational steps are guided not only by the shear stress to which the cell is exposed to by the flow of blood, but also by expression of adhesion molecules, the most important among them are ICAM-1 and VCAM-1 and their integrin counterparts LFA-1 and VLA-4, expressed by the immune cell. These proteins are crucial not only in a mechanically anchoring sense, but they also play a part in an intracellular signaling process leading to a change in migrational direction, overall cell affinity and phenotype. Few is known about how all components shape the movement behaviour on a quantitative level, raising questions about hierarchy, affinity and density of the involved proteins. Besides enhancing the general knowledge of the mechanisms of T-cell migration, the role of ICAM-1 and VCAM-1 in various diseases makes this study a promising endeavour. The approach taken in this thesis is to dissect and recompose the important adhesion molecules on a laminal flow chamber to link the cell’s response to them to specific movement properties and answer the questions addressed above.

Masters of Science / Background: Infection with HIV results in gradual loss of immunologic functions, especially those mediated by CD4+T helper cells with consequent impairment of the immune response leading to severe manifestations of periodontal disease. The lower the CD4+T lymphocyte cell count or the higher the level of immunosuppression, the higher the incidence of periodontal disease in those patients will be. Putative periodontopathic bacteria namely Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia, commonly referred to as "red complex", and many other bacterial species have been implicated in the initiation and progression of periodontal disease. Objective: The present study tests the association between different CD4+T lymphocyte levels and "red complex" pathogens using BANA, in HIV-positive patients with chronic inflammatory periodontal disease (CIPD). Methods: 120 HIV-positive patients from the infectious disease clinic at Tygerberg hospital participated in the study with a mean age of 33.3 years. The CD4+T lymphocyte counts were obtained from patient's medical records. The six Ramjford teeth were used for evaluating periodontal clinical parameters such as plaque index, gingival index, periodontal probing depth and clinical attachment loss. Subgingival plaque samples were collected and analyzed by the enzymatic BANA test for the detection of the "red complex". Results: The CD4+T lymphocyte mean level was 293.43cells/mm3. Statistically significant associations were found between CD4+T cell counts and probing depth (p= 0.0434) and clinical attachment loss (p= 0.0268). Significant associations were found between BANA with all the clinical indices (p= <0.05). However no association was found between CD4+T cell counts and BANA. Conclusion: HIV-positive patients show a high prevalence of "red complex" pathogens subgingivally. Immunosuppression seems to favour the colonization of these species, resulting in periodontal disease manifestations.

Introdução: A doação de plaquetas por aférese é um método de coleta que vem aumentando em relevância. Sabe-se que esta técnica apresenta inúmeras vantagens em comparação à doação de sangue total. Observamos que há uma preocupação na qualidade dos hemocomponentes enviados ao paciente, no entanto, não se observam muitas pesquisas em busca do cuidado com o doador. Órgãos como o Food and Drug Administration (FDA) já publicaram normas mais restritivas em relação à doação de plaquetas por aférese, pois pesquisas apontaram uma diminuição de algumas células e proteínas do sistema imunológico em doadores de repetição. Objetivos: Analisar doadores de plaquetas de repetição quanto a parâmetros hematimétricos e quantificação de subpopulações linfocitárias comparando-os com um grupo controle composto por doadores de sangue total que não doam há no mínimo um ano ou doando pela primeira vez e, ainda avaliar se a frequência de doações, o tempo de procedimento e o número de plaquetas doadas influenciam na contagem de leucócitos totais e nas subpopulações de linfócitos. Metodologia: Foram analisados 88 indivíduos em um estudo caso-controle, sendo que o grupo controle (CO) incluído foi de doadores de sangue total que haviam doado pela primeira vez ou haviam doado sangue total há mais de um ano. Os casos (CA) incluídos foram os doadores de repetição de plaquetaférese (quatro ou mais doações no último ano). O pareamento foi feito por sexo e idade. As amostras de sangue periférico foram coletadas em tubos contendo EDTA e analisadas em até 6 horas por citometria de fluxo, através da utilização de anticorpos monoclonais anti-CD3, CD4, CD8, HLADR, CD19 e CD56. Resultados: Foram avaliados 44 pares de doadores (caso vs controle). Destes, 81,8% eram homens, a média de idade dos grupos foi de 46 ±13 anos nos casos e 47 ±11 nos controles. Comparando os dois grupos, observou-se diferença estatisticamente significativa (p<0,05) na média de quantificação de leucócitos absolutos CA= 6476,6/μL vs CO=7115,4/μL (p=0,017), na média de linfócitos absolutos CA= 1862,6/μL vs CO= 2239,2/μL (p=0,007) e nos marcadores: CD3+/CD8+ (absoluto) CA= 437/μL vs CO= 597/μL (p=0,01), CD3+/CD4+(%) CA= 47,3/μL vs CO= 42,77/μL (p=0,007). Conclusões: Neste estudo foi possível observar que há uma diminuição em algumas células linfoides dos doadores de repetição em relação aos doadores convencionais, no entanto essa diferença não tem relevância clínica, demonstrando que os intervalos de doações que estes doadores estão sendo submetidos é adequado. A contagem de plaquetas dos doadores de repetição se mantiveram no decorrer do ano, este dado nos auxilia para mantermos um banco de dados de doadores de repetição com uma quantificação de plaquetas adequada, podendo ser convocado sem risco de ser bloqueado por contagem inferior ao preconizado. / Introduction: The donation of platelets by apheresis as a collection method has lately grown in relevance. This technique presents several advantages when compared to total blood donation. We understand there is a concern about the quality of the hemocomponents that are administered to the patients; however, there are not many researches concerned with caring for the donor. Entities such as the Food and Drug Administration (FDA) have published more restricting regulations regarding the donation of platelets by apheresis, since researches indicate a decrease in some cells and proteins present in the immunological systems of repeat donors. Objectives: To analyze repeat donors of platelets with regards to hematimetric parameters and quantification of lymphocyte sub-populations by comparing them with a control group consisting of total blood donors that have not donated blood for the past year at least or that are donating for the first time. Additionally, to evaluate if the frequency of donations, the duration of the procedure, and the donated platelet counts influence in the total leukocyte counts and in the sub-populations of lymphocytes. Methodology: We analyzed 88 individuals in a control case study. The control group (CG) consisted of total blood donors in their first donation or that had donated for the last time more than a year before. The cases (CA) included were the repeat donors by platelet apheresis (four or more donations in the past year). We matched the individuals by gender and age. Peripheral blood samples were collected in tubes containing EDTA and analyzed up until 6 hours later by flow cytometry, through monoclonal antibodies anti-CD3, CD4, CD8, HLADR, CD19, and CD56. Results: 44 pairs of donor were evaluated (case vs control). Among them, 81.8% were men, the average age of the groups was 46 (±13) years in the cases and 47 (±11) in the controls. When comparing the two groups, we observed a statistically significant difference (p<0,05) in the average of the quantification of absolute leukocytes CA= 6476.6/μL vs CG=7115.4/μL (p=0.017), in the average of absolute lymphocytes CA= 1862.6/μL vs CG= 2239.2/μL (p=0.007), and in the markers: CD3+/CD8+ (absolute) CA= 437/μL vs CG= 597/μL (p=0,01), CD3+/CD4+(%) CA= 47.3/μL vs CG= 42.77/μL (p=0.007). Conclusions: We were able to note in this study that there is a significant decrease in some lymphoid cells of repeat donors when compared to conventional donors. This difference, however, is not clinically relevant, which demonstrates that the donation intervals to which the donors are subject are appropriate. Platelet numbers of repeat donors remained the same throughout the year. This piece of data helps us keep a database of repeat donors with an adequate platelet number. These donors can be called for without risking of their being blocked in the screening for a number lower than the recommended.

Introduction Multiple sclerosis (MS) is a chronic inflammatory demyelinating and neurodegenerative disease affecting the central nervous system. Although research on the diagnosis and treatment of this serious illness has made significant advances, the MS etiology remains unknown. Although there is still much to be found in the MS pathogenetic mechanisms, understanding the mechanisms of immune-mediated damage to the central nervous system (CNS) components of MS allows not only the introduction of new drugs that positively modulate inflammatory inflammation but also the understanding of immune- mediated diseases affecting the CNS in a broader sense. Methods The Ph.D. thesis is a general introduction followed by an annotated set of author's publications on proteomics, lymphocytic population in MS and disability. Study 1: Proteomic analysis of cerebrospinal fluid for relapsing-remitting multiple sclerosis and clinically isolated syndrome was conducted to find changes in the low molecular cerebrospinal fluid (CSF) segment and to determine what native peptides and how much they are in cerebrospinal fluid among patients with clinically isolated syndrome (CIS) and relapsing-MS (RR MS) compared to a healthy population. Study 2: Lymphocytes in the treatment with interferon beta-1b targeted individual...