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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2014.tde-24022015-091451
Document
Author
Full name
Mariana Pereira Pinho
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Barbuto, Jose Alexandre Marzagao (President)
Câmara, Niels Olsen Saraiva
Chammas, Roger
Title in Portuguese
Enriquecimento antigênico de linhagens tumorais: estratégia para abordagens imunoterapêuticas personalizadas.
Keywords in Portuguese
Câncer
Células dendríticas
Imunoterapia
RNA mensageiro
Vacinas
Abstract in Portuguese
Células dendríticas (DCs) são células apresentadoras de antígenos usadas em estratégias imunoterapêuticas, como as que utilizam híbridos de DCs e células tumorais. Este projeto avaliou a possibilidade de utilizar, como parceiro de fusão das DCs, células de linhagem tumoral previamente transfectadas com mRNA amplificado de células tumorais contra as quais se pretende induzir resposta. Os híbridos foram capazes de induzir uma resposta antígeno-específica e foi possível enriquecer uma célula com antígenos de outra, uma vez que células transfectadas passaram a apresentar mRNAs da célula doadora, e expressar GFP quando a célula doadora era positiva para GFP. Foi possível amplificar integralmente o mRNA para GFP e amplificar os mRNAs de uma célula, gerando um material contendo mRNAs do preparado inicial mas incapaz de aumentar a expressão das moléculas avaliadas nas células transfectadas, mostrando que o protocolo ainda precisa ser aperfeiçoado. Em conjunto, os resultados mostraram que a estratégia de imunoterapia aqui explorada é promissora e merece maior investigação.
Title in English
Antigenic enrichment of tumor cell lines: a strategy for personalized immunotherapeutic approaches.
Keywords in English
Cancer
Dendritic cells
Immunotherapy
Messenger RNA
Vaccines
Abstract in English
Dendritic cells (DCs) are antigen presenting cells widely used in immunotherapy strategies, as the ones that utilize dendritic cell tumor cell hybrids. The present project evaluated the possibility of using cells from tumor cell lines, which were previously transfected with mRNA amplified from tumor cells against which a response is aimed, to fuse with DCs. The hybrids were able to induce a specific immune response. Also, it was possible to enriched one cell with antigens from another one, since transfected cells increased the amount of mRNAs from the donator cell, and expressed GFP when the donator cells expressed this protein. It was possible to successfully amplify the GFP specific mRNA. The mRNAs amplified from RNA of different cell lines contained mRNAs that were present in the total extracted RNA, but were not able to increase the expression of the molecules we attempted to detect in the transfected cells, showing that the protocol still need to be improved. Together, these results show that this new strategy is promising and deserves further investigation.
 
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Publishing Date
2015-02-24
 
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