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Previous issue date: 2016-03-16 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Hsp70 (Heat shock protein 70 kDa) is a chaperone protein which has intra- and extracellular functions. In the early studies, only intracellular functions were operated, as to aid in folding, prevent aggregation and protein refolding. In 80?s a study has shown that this protein can be exported into the extracellular environment exerting effects on other cells, such as the immune system. Our group are exploring the role of this protein in key immune response cells such as macrophages and dendritic cells. Present study noted that the Hsp70 of Mycobacterium tuberculosis can polarize macrophages to a M2 phenotype and this response is dependent on IL-10. Peritoneal and bone marrow derived macrophages were treated with DnaK (Hsp70 prokaryotic homolog) from M. tuberculosis for 24 hours. After this period there was an increase of M2 classic phenotype markers, such as FIZZ1, YM1, CD206, IL-10 and Arg1. On the other hand, there was a decrease in expression of IL-6, MCP-1, TNF-?, NO, MHC class II and CD86 (M1 markers), and these cells are able to promote tumor growth in allogeneic model. It was also observed with KO cells or blocking IL-10R that this modulation is dependent on IL-10. In addition, this cellular modulation is not restricted to macrophages; such protein acting on dendritic cells by modulating them with an immature phenotype, as has been shown in prior works of our group (Borges et al, 2010;. Motta et al., 2007 ). However, it is not yet known what specific region of the protein is essential for that purpose and whether it is restricted to that homologues, DnaK from M. tuberculosis. Therefore, another objective was to express and purify in the same system their counterparts Hspa1a (murine Hsp70) and HSPA1A (human Hsp70) and subsequently testing them in parallel in immunological experimental models. Work is currently in progress with regard to purification of Hspa1a and initial dendritic cells activation state tests were conducted. It is believed that whether there is an effect with DnaK homologues, this effect is less obvious, according to previous results. / A Hsp70 (Heat shock protein 70 kDa) ? uma prote?na de choque t?rmico de 70 kDa que possui fun??es intra e extracelulares. Nos estudos iniciais, somente as fun??es intracelulares eram exploradas, como o aux?lio no dobramento, impedir a agrega??o e o redobramento prot?ico H? d?cadas mostrou-se que essa prote?na pode ser exportada para o ambiente extracelular e exercer efeitos sobre outras c?lulas, como as do sistema imune. O nosso grupo v?m explorando o papel dessa prote?na em c?lulas chave da resposta imune, como macr?fagos e c?lulas dendr?ticas. O presente trabalho observou que a Hsp70 de Mycobacterium tuberculosis pode polarizar macr?fagos a um fen?tipo M2 e que essa resposta ? dependente de IL-10. Macr?fagos peritoneais e derivados de medula ?ssea foram tratados com DnaK (hom?logo procari?tico da Hsp70) de Mycobacterium tuberculosis por 24h. Ap?s esse per?odo observou-se o aumento de marcadores cl?ssicos de fen?tipo M2, como: FIZZ1, YM1, CD206, IL-10, Arg1. Por outro lado, houve uma diminui??o na express?o de IL-6, MCP-1, TNF-?, NO, MHC de classe II e CD86, al?m de essas c?lulas serem capazes de promover o crescimento tumoral em modelo alog?nico. Observou-se tamb?m, com c?lulas KO ou bloqueando o IL-10R que essa modula??o ? dependente de IL-10. Al?m disso, essa modula??o celular n?o ? restrita aos macr?fagos, podendo essa prote?na agir sobre c?lulas dendr?ticas, modulando-as a um fen?tipo imaturo, como j? mostrado em trabalho anteriores do grupo (1,2). Por?m, n?o se sabe ainda qual regi?o espec?fica da prote?na ? essencial para esse efeito e se ele ? restrito a esse hom?lo, DnaK de M. tuberculosis. Portanto, outro objetivo do trabalho foi expressar e purificar no mesmo sistema os seus hom?logos Hspa1a (Hsp70 murina) e HSPA1A (Hsp70 humana) e posteriormente, test?-las em paralelo em modelos experimentais imunol?gicos. O trabalho est? em andamento atualmente em rela??o a purifica??o da Hspa1a e os primeiros testes de estado de ativa??o de c?lulas dendr?ticas foram realizados. Acredita-se que se h? um efeito dos hom?logos de DnaK, esse efeito ? menos evidente, segundo resultados pr?vios.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2.pucrs.br:tede/6875 |
| Date | 16 March 2016 |
| Creators | Lopes, Rafael Lisboa |
| Contributors | Bonorino, Cristina Beatriz Cazabuena |
| Publisher | Pontif?cia Universidade Cat?lica do Rio Grande do Sul, Programa de P?s-Gradua??o em Biologia Celular e Molecular, PUCRS, Brasil, Faculdade de Bioci?ncias |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da PUC_RS, instname:Pontifícia Universidade Católica do Rio Grande do Sul, instacron:PUC_RS |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 8198246930096637360, 600, 600, 600, 600, 36528317262667714, -1634559385931244697, 2075167498588264571 |
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