Made available in DSpace on 2014-06-11T19:27:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2001-02-02Bitstream added on 2014-06-13T19:15:42Z : No. of bitstreams: 1 paes_rcs_me_jabo.pdf: 1518593 bytes, checksum: 4c347db640a4ee036b567916847c5278 (MD5) / A pesquisa sorológica usando Imunodifusão em Gel de Ágar (IOGA) e ELISA Bloqueio de Fase Líquida (BFL-EUSA) foram aplicadas nesse estudo no período de 1996 a 1998, para detectar ou titular, respectivamente, anticorpos contra o antígeno VIA (Virus Infection Associated) e as estirpes de referência 01 Campos, A24 Cruzeiro e C3 Indaial, em algumas espécies silvestres e domésticas susceptíveis e não vacinadas. Do total de 383 amostras sanguíneas colhidas, 78 amostras eram de bovinos de vida livre; 39 de búfalos indianos; 139 de ovinos; 63 de suínos domésticos; 49 de porcos do mato e 15 de cervídeos. Todas as espécies investigadas, exceto os cervídeos, apresentaram anticorpos contra o Vírus da Febre Aftosa (VFA). O teste de IDGA detectou 47 (12,3%) amostras positivas ao antígeno VIA, principalmente em bovinos baguás e búfalos. O BFL-ELISA identificou 175 (45,7%) soros com presença de anticorpos a uma das três estirpes virais; Foi encontrado um maior número de amostras positivas contra a estirpe C3 Indaial bem como maiores títulos, em bovinos baguás. Os bubalinos e ovinos apresentaram uma maior reatividade contra as estirpes A24 Cruzeiro e 01 Campos, respectivamente, demonstrando maior número de soros reagentes. No entanto, ambas as espécies revelaram maiores títulos para a estirpe C3 Indaial. Nos suínos domésticos e silvestres, predominou a reatividade a estirpe A24 Cruzeiro. Em bovinos, a maior ocorrência de soros positivos ao teste de IOGA foi encontrada em animais entre 12 a 36 meses de idade, e na prova BFL-ELISA, em animais acima de 36 meses. Nas demais espécies, ambos os testes laboratoriais utilizados demonstraram um maior número de soros reativos em indivíduos adultos... / A serologic survey, using Agar Gel Immunodifusion (AGID) and liquid Phase Blocking-ELISA (LPB-ELISA) were assessed in this study, in the period from to 1996 to 1998, to detect or titer, respectively, antibodies to Virus Infection Associated Antigen (VIAA) and the foot-and-mouth disease virus (FMDV) 01 Campos, A24 Cruzeiro e C3 Indaial reference strains, in non-vaccinated domestic and wild susceptible animal species. A total of 383 blood serum samples were collected from free-ranging cattle (78 samples), water buffalo (39 samples), sheep (139 samples), domestic (63 samples) and wild pigs (49 samples) and deers (15 samples), living in the Pantanal (Nhecolândia region) of Mato Grosso do Sul State, Brazil. All the investigated species, excepting deer, showed positive animais to FMDV antigens. The AGID detected a total of 47 (12,3%) positive animais for VIA antigen, mostly of them were free-living cattle and buffalo. The LPD-EUSA identified 175 (45,7%) serum samples as reactive to FMDV antigens, specially among cattle, buffalo and sheep. The predominant reactivity to FMDV antigens, characterized by the higher number of positives and the higher antibodies titers, were detected to C3 Indaial strain, particularly among the free-Iiving cattle. Buffaloes and pigs showed, in turn, a prominent reactivity to A24 Cruzeiro strain, while sheep reacted more to 01 Campos strain, though buffalo and sheep serum samples had higher antibody titers to C3 Indaial strain. The highest frequency of positive animais to VIA antigen were found in free-ranging cattle, whose age ranged from 12 to 36 months, contrasting to the results of LPB-EUSA that detected the highest rate of positives in the older bovines (~36 months). The older animais from other species demonstrated, in both serological tests, the highest frequency of positivity... (Complete abstract, access undermentioned eletronic address)

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Single-particle tracking (SPT) is a method used to study the diffusion of various molecules within the cell. SPT involves tagging proteins with optical labels and observing their individual two-dimensional trajectories with a microscope. The analysis of this data provides important information about protein movement and mechanism, and is used to create multistate biological models. One of the challenges in SPT analysis is the variety of complex environments that contribute to heterogeneity within movement paths. In this thesis, we explore the limitations of current methods used to analyze molecular movement, and adapt analytical methods used in animal movement analysis, such as correlated random walks and first-passage time variance, to SPT data of leukocyte function-associated antigen-1 (LFA-1) integral membrane proteins. We discuss the consequences of these methods in understanding different types of heterogeneity in protein movement behaviour, and provide support to results from current experimental work. / Applied Mathematics

Nectine-4 est une molécule d'adhérence membre de la superfamille des immunoglobulines. Elle est localisée au niveau des jonction adhérentes. Exprimée durant l'embryogenèse, nectine-4 n'est pas retrouvée dans les tissus adultes excepté la peau. Des mutations survenant au niveau du gène de la nectine-4 cause le syndrome EDSS affectant le développement de la peau. Nous avons participer à la démonstration que l'expression de nectine-4 est marqueur des carcinomes mammaires,pulmonaires et ovariens. Ainsi, nous avons étudier le role fonctionnel de l'expression de nectine-4 sur la progression tumorale. Les résultats obtenus suggèrent que nectine-4 représente une cible intéressante dans le cadre d'une stratégie d’immunothérapie anti-tumorale.Par l'utilisation d'une méthode multiplex combinant des essais biochimiques, cellulaires et algorithmiques, 5 peptides potentiellement antigéniques ont été identifiés. Toutefois,après génération de lymphocytes cytotoxiques HLA-A*0201 spécifique à partir de PBMC de donneurs sains et la mise en place" de tests de cytotoxicité dirigée; nous avons identifié un nouveau peptide antigénique produit et présenté de façon naturelle à la surface de cellules tumorales. Ce dernier est le peptide nectine-4 N°145 (VLVPPLPSL). En plus de l'identification d'un peptide antigénique, nous avons développé un anticorps monoclonal anti-nectine-4 qui a la capacité de réduire de façon spécifique et significative les capacités métastatiques des cellules tumorales nectine-4 positive.Ainsi, dans cette étude nous avons démontré que nectine-4 qui est un nouveau antigène associé aux tumeurs, affecte la progression tumorale, mais aussi il est possible de cibler cette molécule par des stratégies d'immunothérapie car nous avons pu identifiés un peptide antigénique et un anticorps monoclonal. L'efficacité d'une stratégie d'immunothérapie est en cours de réalisation actuellement. / Nectin-4 is a cell surface adhesion molecule, member of the Ig-superfamily, and is localized at adherens junctions. Nectin4 is expressed during embryogenesis but not in adult tissues, except in the skin. Mutations in nectin-4 gene in humans cause the EDSS syndrome that affects skin development (EctoDermal and Syndactly Syndrome). We and others have recently demonstrated that nectin-4 expression was a tumoral marker of breast, lung and ovarian carcinoma. We thus started to investigate the functional role of nectin-4 overexpression in breast cancer. Using in vitro and in vivo assays, the preliminary results demonstrate that nectin-4 increased the tumorigenicity of malignant cells.Altogether, these results suggest that nectin-4 might be a candidate target forimmunotherapy (vaccination and antibody based therapy). Using a multiplex approachbased on biochemical, cellular and algorithmic assays, five relevant nectin-4 epitopes were identified. Specific cytotoxic T lymphocyte (CTL) populations from healthy donors that recognized and lyzed peptide-pulsed HLA-A*0201 tumor cells were identified. HLAA*0201-restricted CTL that recognized the N4-145 (VLVPPLPSL) nectin-4 epitope was characterized extensively. This CTL kills breast tumor cells that express nectin4, strongly demonstrating that this peptide could be naturally processed and recognized by specific CTL. In parallel, we also tested a blocking monoclonal antibody against the extracellular region of nectin-4. We next demonstrated that this antibody reduced the metastatic capacity of tumors expressing nectin4. To summarize, in this study, we identified nectin-4 as a newcell surface Tumor Associated Antigen and demonstrated its likely implication in cancertumorigenesis. In parallel, we have developed the specific tools required to conduct an effective immunotherapy targeting nectin4 over-expressing cells, which are currently under investigation.