Open Access Theses and Dissertations

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Universidade do Rio Grande do Sul

1. Cibulski, Samuel Paulo. Saponinas de Quillaja brasiliensis: potencial imunoadjuvante e mecanismos celulares e moleculares de ação.

Degree: 2015, Universidade do Rio Grande do Sul

URL: http://hdl.handle.net/10183/127089 

A formulação de vacinas efetivas frequentemente requer a adição de adjuvantes capazes de otimizar as respostas imunes humoral e celular. Com o objetivo principal de contribuir para o desenvolvimento de novos adjuvantes, este trabalho foi desenvolvido buscando aprofundar o conhecimento do mecanismo de ação imunoadjuvante de preparações de saponinas de Quillaja brasiliensis e suas formulações em complexos imunoestimulantes do tipo ISCOM. Como a toxicidade das saponinas é um fator crítico para seu uso em preparações vacinais, inicialmente foram realizados ensaios visando comparar a toxicidade in vitro e in vivo de saponinas extraídas de Quillaja brasiliensis com saponinas de ação imunoestimulante reconhecidas, extraídas de Quillaja saponaria (Quil A). O potencial imunoadjuvante das saponinas solúveis de Q. brasiliensis foi avaliado utilizando preparações com dois antígenos: ovalbumina (OVA) e vírus da diarreia viral bovina (BVDV). Numa etapa seguinte, a atividade imunoadjuvante de ISCOMs preparados com saponinas de Q. brasiliensis foram avaliadas em duas vias de administração. O potencial imunomodulador dessas saponinas foi verificado em experimentos de recrutamento celular in vivo e expressão de genes relacionados ao sistema imune. Os resultados mostraram que saponinas de Q. brasiliensis são menos tóxicas que as de Quil A e apresentam atividade adjuvante similar, caracterizada por um perfil Th1/Th2 balanceado. Q. brasiliensis promoveu uma forte resposta imune celular do tipo Th1 caracterizada por uma robusta reação de hipersensibilidade celular tardia (DTH) e pela produção de IFN- e IL-2. A resposta imune induzida pelos ISCOMs produzidos a partir de saponinas de Q. brasiliensis foram superiores às respostas induzidas pelas saponinas solúveis. Os testes in vivo mostraram que as saponinas de Q. brasiliensis promovem um ambiente imunocompetente no local da inoculação e nos linfonodos drenantes. Esse ambiente foi caracterizado pelo intenso influxo celular (neutrófilos, células NK, células dendríticas, linfócitos T e B), além da expressão diferencial de genes relacionados à ativação do sistema imune. Em suma, os resultados mostraram que saponinas de Q. brasiliensis são seguras e seus potencial adjuvante foi equivalente a saponinas com ação imunoadjuvante conhecida de Q. saponaria.

Effective vaccine formulations frequently require addition of adjuvants able to optimize the cellular and humoral immune responses. With the goal to contribute to the development of new classes of adjuvants, this work was developed in order to achieve deep knowledge on the imunoadjuvant mode of action for Quillaja brasiliensis saponins incorporated into immunostimulant complex (ISCOM). The toxicity of saponins is a critical factor for its usage as vaccine preparations. At first, in vivo and in vivo citoxicity assays were carried out to compare to the effects between saponins extracted from Quillaja brasiliensis and the immunostimulant saponins already known from Quillaja saponaria (Quil A). Imunoadjuvant potential of soluble saponins from Q.…

Advisors/Committee Members: Roehe, Paulo Michel.

Subjects/Keywords: Saponins; Quillaja brasiliensis : Saponinas; Biologia celular; ISCOMS; Biologia molecular; Hemolysis; Adjuvants; Recrutamento celular; Ativação imune; Cell recruitment; Immune activation

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Cibulski, S. P. (2015). Saponinas de Quillaja brasiliensis: potencial imunoadjuvante e mecanismos celulares e moleculares de ação. (Thesis). Universidade do Rio Grande do Sul. Retrieved from http://hdl.handle.net/10183/127089

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Cibulski, Samuel Paulo. “Saponinas de Quillaja brasiliensis: potencial imunoadjuvante e mecanismos celulares e moleculares de ação.” 2015. Thesis, Universidade do Rio Grande do Sul. Accessed January 16, 2021. http://hdl.handle.net/10183/127089.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Cibulski, Samuel Paulo. “Saponinas de Quillaja brasiliensis: potencial imunoadjuvante e mecanismos celulares e moleculares de ação.” 2015. Web. 16 Jan 2021.

Vancouver:

Cibulski SP. Saponinas de Quillaja brasiliensis: potencial imunoadjuvante e mecanismos celulares e moleculares de ação. [Internet] [Thesis]. Universidade do Rio Grande do Sul; 2015. [cited 2021 Jan 16]. Available from: http://hdl.handle.net/10183/127089.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Cibulski SP. Saponinas de Quillaja brasiliensis: potencial imunoadjuvante e mecanismos celulares e moleculares de ação. [Thesis]. Universidade do Rio Grande do Sul; 2015. Available from: http://hdl.handle.net/10183/127089

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation


University of Gothenburg / Göteborgs Universitet

2. Grdic, Dubravka Kristina 1968-. Evidence for the differential regulation of mucosal IgA immunity and oral tolerance. Studies in gene knockout and transgenic mice using the cholera toxin adjuvant system.

Degree: 1999, University of Gothenburg / Göteborgs Universitet

URL: http://hdl.handle.net/2077/11228 

Despite the continuos exposure to a myriad of food and microbial antigens, inflammatory reactions in the intestinal mucosa are rare. Homeostasis in the intestinal mucosa is maintained by regulatory T cells, which can be tolerized or primed to provide efficient B cell help. Two types of responses can be elicited by mucosal antigen; active IgA immunity or oral tolerance. The latter being the dominant type of reaction to almost all soluble antigens, whereas active IgA immunity requires the use of effective adjuvants, such as cholera toxin (CT) or ISCOMS. This thesis explores the regulatory role of CD4+ and CD8+ T cells for oral tolerance induction and active IgA immunity. Moreover, we have addressed the question of whether oral tolerance and active IgA immunity are reciprocally regulated or can co-exist. The governing functions of IL-12 (Th1) or IL-4 (Th2) for induction of tolerance or active IgA immunity were also assessed using gene knockout mice and ISCOMS and CT-adjuvant. Finally, we experimentally addressed the hypothesis that the type of response elicited by oral antigen administration is determined at the antigen-presenting cell (APC) level. We found that mice lacking CD8+ T cells exhibited significantly stronger mucosal IgA responses and failed to develop local tolerance to fed antigens, suggesting a down-regulatory function of CD8+ T cells in the normal gut mucosa. By contrast, oral tolerance at the systemic level, was clearly independent of CD8+ T cells, but affected both Th1 and Th2 CD4+ T cell functions. Furthermore, already established tolerance could not be abrogated by CT, whereas this adjuvant and ISCOMS prevented the induction of tolerance in all mouse strains tested, including IL-4 and IL-12-deficient mice. Th2 cells /IL-4 was critical for CT-induced IgA immunity, while ISCOMS were dependent on Th1 cells/IL-12 activity, indicating differential regulatory requirements for induction of mucosal IgA immune responses. Oral tolerance was readily induced in both these mouse strains in the absence of adjuvant, suggesting also separate regulatory pathways for induction of oral tolerance and active IgA immunity. The reciprocal regulation of these response patterns was further indicated by the finding that antigen conjugated to CT, strongly promoted mucosal IgA responses even in IL-4 deficient mice, while oral tolerance was prevented by the conjugation. The ability to circumvent the requirement for IL-4 was dependent on the enzymatic activity of CT, acting on the APC, because conjugation to the enzymatically inactive, but cell-receptor binding CTB, failed to affect IgA immunity. Using OVA TCR transgenic mice we could demonstrate that clonal T cells can be either tolerized or induced to enhance active IgA immunity depending on the function of the APC in the gut and that fed antigen can simultaneously induce local IgA immunity and systemic CD4+ T cell tolerance, arguing for the existence of separate and distinct activation pathways of the two response patterns; oral tolerance and active IgA immunity, in the gut…

Subjects/Keywords: Oral tolerance; mucosal immunity; knockout mice; CT; ISCOMS. PAGE

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Grdic, D. K. 1. (1999). Evidence for the differential regulation of mucosal IgA immunity and oral tolerance. Studies in gene knockout and transgenic mice using the cholera toxin adjuvant system. (Thesis). University of Gothenburg / Göteborgs Universitet. Retrieved from http://hdl.handle.net/2077/11228

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Grdic, Dubravka Kristina 1968-. “Evidence for the differential regulation of mucosal IgA immunity and oral tolerance. Studies in gene knockout and transgenic mice using the cholera toxin adjuvant system.” 1999. Thesis, University of Gothenburg / Göteborgs Universitet. Accessed January 16, 2021. http://hdl.handle.net/2077/11228.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Grdic, Dubravka Kristina 1968-. “Evidence for the differential regulation of mucosal IgA immunity and oral tolerance. Studies in gene knockout and transgenic mice using the cholera toxin adjuvant system.” 1999. Web. 16 Jan 2021.

Vancouver:

Grdic DK1. Evidence for the differential regulation of mucosal IgA immunity and oral tolerance. Studies in gene knockout and transgenic mice using the cholera toxin adjuvant system. [Internet] [Thesis]. University of Gothenburg / Göteborgs Universitet; 1999. [cited 2021 Jan 16]. Available from: http://hdl.handle.net/2077/11228.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Grdic DK1. Evidence for the differential regulation of mucosal IgA immunity and oral tolerance. Studies in gene knockout and transgenic mice using the cholera toxin adjuvant system. [Thesis]. University of Gothenburg / Göteborgs Universitet; 1999. Available from: http://hdl.handle.net/2077/11228

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

.