Thesis (M.S.)--University of Wisconsin--Madison, 1965. / eContent provider-neutral record in process. Description based on print version record. Bibliography: l. 58-62.

Thesis (M. Phil.)--University of Hong Kong, 1998. / Includes bibliographical references (leaves 100-128).

Thesis (M. Phil.)--University of Hong Kong, 1996. / Includes bibliographical references (leaves 162-185).

Thesis (Ph. D.)--University of Chicago, Dept. of Pathology, June 2003. / Includes bibliographical references. Also available on the Internet.

Thesis (Ph. D.)--University of Wisconsin, 1966. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (M.S.)--Northern Michigan University, 2009. / Bibliography: leaves 38-42.

Banca: Mario Tsunezi Shimizu / Banca: Olga Fichman / Banca: Esther Birman / Banca: Luiz Jacinto da Silva / Banca: Terezinha de Oliveira Nogueira / Resumo: Anticorpos naturais para antígenos próprios e exógenos foram analisados pela técnica de ELISA, em 60 soros de pacientes cronicamente infectados pelo T. cruzi e, em 40 soros de indivíduos sofrendo de uma micose profunda, freqüentemente encontrada na América Latina (Paracoccidioimicose). Cerca de 80% dos indivíduos do primeiro grupo tinham níveis significativamente elevados de anticorpos antilaminina (M=4,75, DP±1,88) quando comparados com os controles saudáveis e, diferentes especificidades de anticorpos estavam associadas com antilaminina nos soros chagásicos. Foram observados anticorpos reativos com as proteínas do citoesqueleto, especialmente com a banda 3 e seus peptídeos, como o de 62 kDa. Por meio de cromatografia, em Proteína-A - Sepharose, mostramos que uma parte significante do antiGal natural pode estar ligada por sua região Fab a outra molécula de imunoglobulina e/ou a sítios alternativos de ligação na Proteína A. O achado de lgG antiGal nos imunocomplexos reforçam a primeira hipótese. Todavia, é possível que algumas lgG aniiGal pertençam ao subgrupo VH111 de imunoglobulinas. Entre os 40 soros de PCM examinados, a maioria tinha níveis normais de anticorpos naturais. Todavia, 30% dos pacientes crônicos apresentavam aumentos de pelo menos uma especificidade de anticorpo natural, tal como antiactina, antimiosina e antiGal. Na forma juvenil os anticorpos antiactina estavam aumentados de 2,4 vezes (1,9 a 5,3 vezes), sugerindo uma associação com a doença disseminada. lmunocomplexos foram pesquisados nos soros de PCM utilizando a técnica de precipitação com polietileno glicol 6000. Nos imunocomplexos Isolados foram encontrados principalmente anticorpos específicos para antígenos solúveis de P. brasiliensis / Abstract: Using ELISA technique, natural antibodies against self and non self antigens were determined in 80 patients chronically intected by T. cruzi and 40 individuals suffering from a deep mycosis frequentely found in Latin Amarica (Paracoccidioidomycosis - PCM). Two forms of PCM were investigated: adult forms and juvenil type of disease. Eighty percent (80%) of the former group had significantly elevated anti-laminin antibody levels (M=4.7,SD±1.8) compared with healthy controls and different specificities of antibody were associated with anti-laminin in pathological sera. A notable binding to cytoskeletal proteins was observed, specially with band 3 and their peptides derivates, such as 62 kDa peptide. By means of Protein A chromatography we were able to show that natural anti-Gal antibodies may be bound by their Fab region to other immunoglobulins and/or to Protein A by alternative sites of binding. The finding of lgG anti-Gal antibodies in circulating immune complexes isolated from chagasic sera supported the first alternative. However, it is possible that some of lgG anti-Gal antibodies, belong to VH111 subgroup of immunoglobulins, that bind directly to Protein A. Among the 40 sera from PCM examined, the majority was considered as not exhibiting a signilicantly higher binding than normal sera to antigens tested. However thirty percent (30%) of the chronic patients had an increased levels of natural antibodies at least for one specificity such as actyn, myosin and Gala1,3Gal epitopes. ln juvenil type of PCM the mean value found for actyn was also increased 2,42 (range 1,0 to 5,3). Utilizing the polyethylene glicol precipitation the presence of circulating immune complexes was investigated in PCM sera. Specific antibodies for soluble antigens from P. brasiliensis and natural antibodies against myoglobin, myosin and Gala1,3 Gal epitopes were characterized

Bovine tuberculosis (BTB) is a chronic infectious disease caused by the bacterium Mycobacterium bovis. It infects animals and can be a source of zoonosis. In the last five years, Canada has faced two outbreaks of bovine tuberculosis in the years 2018 and 2016. BTB is mainly diagnosed using the Tuberculin skin test, a test that detects the cellular immune response to administered purified protein derivative (PPD). A drawback of this test is the high level of false-positive test results caused by the immune response to PPD proteins that are conserved in non-tuberculous mycobacteria. Current serodiagnostic tests can detect the disease especially in the advanced state with, low sensitivity ranging between 9-45%. It is hypothesized that the profiling of the humoral immune responses to selected proteins will lead to the discovery of novel immunogenic protein antigens for improved BTB diagnostic tests. Bioinformatic tools used in this research for the prediction of extracellular or outermembrane proteins from M. bovis genome sequences identified 96 protein candidates. Also, a proteomic study conducted to identify proteins secreted by M. bovis, together with the review of previous proteomic studies of the PPD, led to the identification of an additional 92 protein candidates. A high throughput system was developed to efficiently express and analyze these antigens involving the PCR amplification, in vivo cloning, and in vitro expression of open reading frames (ORFs) coding for selected protein candidates. This was followed by a high throughput recombinant protein purification and a microarray analysis of these purified recombinant proteins with sera from M. bovis-infected animals. The system successfully amplified, cloned, and expressed 159 recombinant proteins. From the microarray screening, 13 antigens exhibited immunological reactions and were able to differentiate the sera of the infected animals from the controls. Out of those 13 antigens, 4 novel antigens Mb2740c, Mb0598c, Mb3469c, and Mb3453c, in addition to two well-known antigens MPB70 and MPB83, gave the highest reactivity; they were selected for further evaluation for diagnostic applications using ELISA and dot blot assays. Two antigens Mb3469c and Mb3453c had a significant ability to differentiate between infected and control cattle as tested in ELISA and dot blot assays, respectively and demonstrated by ROC curve analysis. These two novel antigens could be added to the panel of serodiagnostic antigens for improving BTB serodiagnosis and could be beneficial in the detection of outbreaks caused by certain M. bovis strains.

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