Thesis (M.S.)--Virginia Polytechnic Institute and State University, 1992. / Vita. Abstract. Includes bibliographical references (leaves 33-36). Also available via the Internet.

Thesis (M.S.)--University of Wisconsin--Madison, 1966. / eContent provider-neutral record in process. Description based on print version record. Bibliography: l. 77-84.

For many years bovine brucellosis was a zoonosis endemic in large parts of the world. While it is still endemic in some parts, such as the Middle East or India, several countries such as Australia and Canada have successfully eradicated brucellosis in cattle by applying vaccines, improving the hygienic standards in cattle breeding, and slaughtering or quarantining infected animals. The large economical impact of bovine brucellosis and its virulence for humans, coming in direct contact to fluid discharges from infected animals, makes the eradication of bovine brucellosis important to achieve. To achieve this goal several vaccines have been developed in the past decades. Today the two most commonly used vaccines are Brucella abortus vaccine strain 19 and strain RB51. Both vaccines have been shown to be effective, but the mechanisms of immune responses following vaccination with either of the vaccines are not understood yet. In this thesis we analyze the immunological data obtained through vaccination with the two strains using mathematical modeling. We first design a measure that allows us to separate the subjects into good and bad responders. Then we investigate differences in the immune responses following vaccination with strain 19 or strain RB51 and boosting with strain RB51. We develop a mathematical model of immune responses that accounts for formation of antagonistic pro and anti-inflammatory and memory cells. We show that different characteristics of pro-inflammatory cell development and activity have an impact on the number of memory cells obtained after vaccination. / Master of Science

Brucellosis is a chronic, recurring disease caused by the bacterium Brucella abortus, along with other species of the genus Brucella, and is one of the most common bacterial zoonosis worldwide. The bacteria preferentially infect and reside within host macrophages, causing an undulant fever, joint pain, and other flu-like symptoms, in addition to more severe problems like hepatosplenomegaly and endocarditis. Brucella infection is most often acquired via inhalation through the respiratory route, or via consumption of unpasteurized dairy products. Although ingestion is a major route of infection, the transcriptional response of B. abortus during oral infection remains poorly characterized. In this project, RNA sequencing was used to discover genes with the greatest transcriptional changes in B. abortus subjected to deoxycholate, a host bile acid encountered by bacteria during oral infection. Gene deletion strains of B. abortus were then created and tested for susceptibility to pH and bile acid stress, along with their ability to invade and replicate within macrophages. If the genes of interest are important for the oral infection process, B. abortus strains lacking these genes will likely be more susceptible to pH and deoxycholate stress and may exhibit attenuation in the macrophage infection model. Determination of genes important for the oral infection process would further elucidate the molecular mechanisms by which B. abortus invades the host, and could help lead to future treatments and novel therapeutics. / Master of Science

The roles of genes responsible for the survival and persistence of Brucella in the host and the relationship between these genes and the disease were investigated via signature-tagged transposon mutagenesis. As much as 8% of the Brucella genome is important for survival of this organism in the host. This is an unusually high number and may help to explain the chronic or persistent nature of Brucella infections. Mutants attenuated in the mouse model were divided into two groups. The early mutants failed to establish infection or colonize the host. The late mutants colonized the host but failed to maintain infection. The vaccine potential of two mutants (virB10 and gcvH) that were unable to sustain infection was compared to that of a vaccine strain, S19. Survival of strain S19 in vivo was up to 12 weeks while virB10 and gcvH mutants were cleared from spleen at 8, and 24 weeks post-inoculation, respectively. Mice were vaccinated with individual mutants and then challenged with virulent S2308 at 8, 16, and 24 weeks postvaccination. As a result, protective immunity correlated with persistence of the mutant strain [gcvH>virB10]. These results suggest that survival is one of several factors that may influence protective immunity making it difficult to compare strains. For example, examination of host immune response revealed a similar pattern of host immune function (TH1 over TH2) in all mice except those vaccinated with virB10 mutant. Since gcvH mutant provided the best immunity, experiments were designed to explore its contribution of persistence to protection. In an effort to reduce non-specific activation induced by prolonged survival of gcvH mutant, protection was monitored after different periods of vaccination exposure followed with doxycycline treatment. In these studies, persistence of gcvH mutant enhanced protection against challenge. Overall, defined mutations in genes affecting survival may render mutants as vaccine candidates capable of stimulating protective immunity equal to or better than fortuitously isolated attenuated strains. Future studies should focus on characterization of these and other genes responsible for the persistence of Brucella to improve the safety and efficacy of live vaccines.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2002. / Härtill 5 uppsatser.

Thesis (M.S.)--University of Wyoming, 2008. / Title from PDF title page (viewed on August 4, 2009). Includes bibliographical references.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1960. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 71-75).

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, 2010. / Submitted by Luanna Maia (luanna@bce.unb.br) on 2011-06-06T15:36:47Z No. of bitstreams: 1 2010_AnaPaulaPaivadeFaria.pdf: 488721 bytes, checksum: 3bda6a7be195d840ac115ddaae3b5983 (MD5) / Approved for entry into archive by Luanna Maia(luanna@bce.unb.br) on 2011-06-06T15:37:31Z (GMT) No. of bitstreams: 1 2010_AnaPaulaPaivadeFaria.pdf: 488721 bytes, checksum: 3bda6a7be195d840ac115ddaae3b5983 (MD5) / Made available in DSpace on 2011-06-06T15:37:31Z (GMT). No. of bitstreams: 1 2010_AnaPaulaPaivadeFaria.pdf: 488721 bytes, checksum: 3bda6a7be195d840ac115ddaae3b5983 (MD5) / O atual estudo objetivou analisar a variabilidade genetica de linhagens vacinais de Brucella, avaliando ainda os efeitos de repiques sucessivos sobre as mesmas. Trata-se de doenca zoonotica de alcance mundial e que causa prejuizos economicos na producao animal e danos a saude publica. Visando controlar a doenca, foi instituido o Programa Nacional de Controle e Erradicacao da Brucelose e Tuberculose Animal em 2001, que inclui a vacinacao de femeas. Quanto a avaliacao genetica de linhagens vacinais, a tecnica de analise em multilocus das repeticoes em tandem de numero variavel - MLVA permite identificar a variabilidade genetica microbiana de forma acurada, identificando-se genogrupos de isolados. O atual estudo analisou 63 amostras de linhagens vacinais B19 (n=53) e RB51 (n=10), provenientes de sete laboratorios brasileiros. Tambem foi avaliado o efeito de sucessivos repiques sobre a o genoma microbiano. Desta forma, foram identificados quatro genogrupos distintos. No genogrupo I foram agrupadas as 10 amostras de B19 do laboratorio G e seus 10 repiques. Quarenta e tres amostras de B19 dos laboratorios A, B, D, E, F, H, bem como os 10 repiques das amostras E e da cepa de referencia S (USDA) foram agrupadas no genogrupo II. O genogrupo III foi composto pelas amostras vacinais de RB51 do laboratorio Y e pelos 10 repiques da linhagem Y1. Finalmente, o genogrupo IV incluiu oito amostras vacinais de RB51 e os 10 repiques das amostras X1 e da cepa de referencia Schurig. As linhagens vacinais B19 e RB51 derivaram de ramificacoes geneticas distintas que formaram, cada qual, dois genogrupos, com divergencia apenas nos loci Bruce07 e Bruce43. Concluindo, a tecnica de MLVA mostrou adequada capacidade discriminatoria, podendo ser utilizada para analises de qualidade na producao de vacinas comerciais em nosso Pais. Foi observado, ainda, que 10 passagens sucessivas nao afetam os loci avaliados. ______________________________________________________________________________ ABSTRACT / The present study was aimed to analyze the genetic variability among Brucella abortus lineages present in commercial vaccines from Brazilian laboratories. The brucellosis is a worldwide distributed zoonotic disease. The disease may affect the livestock production as well as the human health. To control the spread of Brucellosis in Brazil, our country established the National Program for Controlling and Eradication of Animal Brucellosis and Tuberculosis – PNCEBT which included the vaccination of female bovines. The multiple locus variable number of tandem repeat analysis (MLVA) is a rapid and accurate method used to access the genetic variability among microbial isolates and to discriminate groups of genetically-related isolates. We used a previously described 16 loci MLVA method to analyze a sample composed by B19 vaccines (n=53) and RB51 vaccines (n=10) from seven Brazilian laboratories. The genetic variability after 10 successive cultures was also accessed. We identified four distinct genogroups of isolates. The genogroup I contained ten B19 samples from the laboratory G and their respective successive cultures. Forty-three B19 samples of the laboratories A, B, D, E, F, and H as well as the successive cultures of the reference S (USDA) strain and the commercial strain E, were all clustered into the genogroup II. The genogroup III included the RB51 laboratory Y’s samples and the Y1’s successive cultures. Finally, eight RB51 samples, the successive X1’s cultures, and the reference Schurig strain, were clustered into the genogroup IV. Our results revealed that the B19 and RB51 lineages derived from two distinct genetic branches that diverged in the locus Bruce07 and Bruce43. In conclusion, the MLVA showed accurate discriminatory results and would be used in future evaluations of the commercial vaccines of Brucellosis. The results of our study also showed that ten successive cultures were not able to generate genetic variability into the loci studied.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-06-21T19:56:49Z No. of bitstreams: 1 marinacoelhodequeiroz.pdf: 1087028 bytes, checksum: 2f94da92600324bcb482571c6bb64360 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-08-07T19:19:33Z (GMT) No. of bitstreams: 1 marinacoelhodequeiroz.pdf: 1087028 bytes, checksum: 2f94da92600324bcb482571c6bb64360 (MD5) / Made available in DSpace on 2017-08-07T19:19:33Z (GMT). No. of bitstreams: 1 marinacoelhodequeiroz.pdf: 1087028 bytes, checksum: 2f94da92600324bcb482571c6bb64360 (MD5) Previous issue date: 2013-02-08 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A Brucelose é uma zoonose causada por bactérias do gênero Brucella que causa em seres humanos febre ondulante, endocardite, artrite e osteomielite e em animais causa aborto e infertilidade. Destaca-se na resposta imune contra a Brucella a produção de IFN-ꝩ e a atividade de linfócitos T CD8+, destruindo o patógeno ou deixando-o vulnerável aos mecanismos extracelulares da imunidade. A Talidomida (Td) é um fármaco capaz de induzir a formação de um perfil TH1, com elevada produção de IFN-ꝩ, além de co-estimular e induzir a proliferação e produção de citocinas por linfócitos T CD8+. Assim, o objetivo deste estudo foi avaliar a influência do tratamento com Talidomida na resposta imune contra a B. abortus, focando no futuro desenvolvimento de novos fármacos ou componentes vacinais que auxiliem no combate a esta importante patologia. Foram avaliadas a forma de tratamento e a concentração de Talidomida capaz de influenciar a carga bacteriana presente no baço de animais C57BL/6 infectados; O número de bactérias no baço de animais tratados, após 1,3 e 6 semanas de infecção; A produção de IFN-ꝩ e óxido nítrico em esplenócitos derivados destes animais; A produção de citocinas (IL-12, IFN-ꝩ e TNF-α) no tecido hepático; O percentual de células T CD4+ ou CD8+ presentes no baço de animais tratados e infectados e o potencial citotóxico de esplenócitos derivados destes animais. Foi verificado que o tratamento prévio com Td reduziu a carga bacteriana em todas as semanas avaliadas, conforme evidenciado pela cinética. A produção de IFN-ꝩ e óxido nítrico foram maiores em esplenócitos derivados de animais tratados com Td. A produção de IL-12, IFN-ꝩ e TNF-α foi elevada no tecido hepático após o tratamento. E nos esplenócitos foi observado maior percentual de células T CD8+ e maior citotoxicidade. Assim, os resultados encontrados até o momento nos levam a sugerir que o tratamento com Talidomida potencializam a eliminação do patógeno. / Brucellosis is a zoonosis caused by bacteria from the genus Brucella which causes undulant fever, endocarditis, arthritis and osteomyelitis in humans and, in animals, it causes abortion and infertility. The production of IFN-ꝩ and the activity of T CD8+ lymphocytes have an important role in the immune response against Brucella destroying the pathogen or leaving it vulnerable to extracellular immunity mechanisms. Thalidomide (Td) is a drug which is able to induce the formation of a TH1 profile, with high production of IFN-ꝩ, and it also coestimulates and induces proliferation and production of cytokines by T CD8+ lynphocytes. The purpose of this study was to evaluate the influence of the treatment with Thalidomide on the immune response against B. abortus, focusing in the development of new drugs or vaccine components in the future which will help in the fight against this important pathology. The form of treatment and Thalidomide concentration capable of influencing the bacterial load in the spleen of C57BL/6 infected animals were evaluated; the number of bacteria in the spleen of treated animals after 1, 3 and 6 weeks of infection; the production of IFN-ꝩ and nitric oxide in splenocytes from these animals; the (IL-12, IFN-ꝩ e TNF-α) cytokine production in the liver tissue; the percentage of T CD4+ or CD8+ cells in the spleen of treated and infected animals and the cytotoxic potential of splenocytes from these animals. In this study it was verified that previous treatment with Td reduced the bacterial load in all the evaluated weeks, as it was evidenced by kinetics. The production of IFN-ꝩ and nitric oxide was higher in splenocytes from animals treated with Td. The production of IL-12, IFN-ꝩ and TNF-α was high in the liver tissue after treatment. And a higher percentage of T CD8+ cells and higher citotoxicity were observed in splenocytes. The results so far suggest that treatment with Thalidomide potentializes the elimination of the pathogen.