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Master's Dissertation
DOI
10.11606/D.5.2013.tde-20022014-114721
Document
Author
Full name
Thor Andreas Silva Di Sessa
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Taborda, Carlos Pelleschi (President)
Pagliari, Carla
Rodrigues, Elaine Guadelupe
Title in Portuguese
Terapia gênica na paracoccidioidomicose experimental utilizando vetor de expressão de HSP60 E mIL-12
Keywords in Portuguese
Camundongos endogâmicos BALB/c
Chaperonina 60
Homeostase
Paracoccidioides
Terapia gênica
Vacinas/administração e dosagem
Abstract in Portuguese
A paracoccidioidomicose (PCM) é uma doença sistêmica de caráter granulomatoso, causada pelo fungo termodimórfico Paracoccidioides spp. A PCM é endêmica na America Latina e aproximadamente 80% do pacientes vivem no território brasileiro. O tratamento medicamentoso é eficiente, entretanto, é longo e vários pacientes acabam abandonando e recidivas são comuns neste grupo. A utilização de uma vacina terapêutica poderia resultar na redução do tempo de tratamento assim como, recuperar a resposta imune do hospedeiro frente ao fungo. As vacinas de DNA são uma abordagem promissora na imunoterapia e podem ser injetadas por via intramuscular, intradérmica ou via mucosa. As proteínas de choque térmico (HSPs) são proteínas que estão ligadas a homeostase celular e também possuem efeitos imunológicos em diversos casos como doenças infecciosas e autoimunes. No presente trabalho, analisamos o esquema vacinal terapêutico em camundongos BALB/c previamente infectados intratraquealmente com 3x105 leveduras de P. brasiliensis Pb18, 60 dias depois, submetidos a imunização com pcDNA3 contendo sequências codificadoras de PbHSP60 e/ou IL-12 murina e/ou vetor vazio. Foi observada redução significativa no número de unidades formadoras de colônia (UFCs) nos pulmões de camundongos imunizados com PbHSP60. Os grupos que receberam PbHSP60+pcDNA3 vazio ou PbHSP60x2 apresentaram os maiores índices de redução da cargas fúngicas. A inclusão do plasmídeo contendo o inserto de mIL-12, resultou em um efeito deletério. A análise dos cortes histológicos indicou que os animais vacinados apresentavam áreas bem preservadas e com poucos ou nenhum foco de granuloma. Detectamos um perfil de citocinas típico Th1/Th2. Nossos resultados sugerem que a imunização utilizando plasmídeo contendo o inserto HSP60, tem grande potencial vacinal
Title in English
Gene therapy in experimental paracoccidioidomycosis using HSP60 expression vector and mIL-12
Keywords in English
Chaperonin 60
Gene therapy
Homeostasis
Mice inbred BALB /c
Paracoccidioides
Vaccines/administration and dosage
Abstract in English
The paracoccidioidomycosis (PCM) is a systemic granulomatous disease of character, caused by the thermally dimorphic fungus Paracoccidioides spp. The PCM is endemic in Latin America and approximately 80% of patients are living in Brazil. The medical treatment is effective, however, is long and many patients end up abandoning and relapses are common in this group.The use of a therapeutic vaccine could result in the reducing time of treatment as well as recover the host immune response against the fungus. DNA vaccines are a promising approach for immunotherapy and can be injected by intramuscular, intradermal, or mucosal route. The heat shock proteins (HSPs) are proteins that are linked to cellular homeostasis and also have immunological effects in many cases as infectious and autoimmune diseases. In the present study, we analyzed the therapeutic vaccine schedule in BALB/c mice previously infected intratracheally with 3x105 yeast of P. brasiliensis strain 18, and 60 days after, undergoing immunization with pcDNA3 containing coding sequences PbHSP60 and / or murine IL-12 and / or empty vector. Significant reduction was observed in the number of colony forming units (CFU) in the lungs of mice immunized with PbHSP60. The groups that received empty pcDNA3 and PbHSP60 or PbHSP60x2 have higher rates of reduced fungal loads. The inclusion of the plasmid containing the insert mIL-12 resulted in a deleterious effect. The analysis of histological sections indicated that vaccinated animals had wellpreserved, with few or no focus of granuloma areas. It was detected a profile typical Th1/Th2 cytokines. Our results suggest that immunization using plasmid containing the insert HSP60 vaccine has great potential
 
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Publishing Date
2014-02-24
 
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