Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocols
Autor(a) principal: | |
---|---|
Data de Publicação: | 2019 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10773/29894 |
Resumo: | Dendritic cells (DCs) are cells specialized in antigen presentation that play an important role in the connection between innate and adaptive immunity. These cells have the unique ability to recognize, capture, process and present antigens to naïve T cells, leading to their polarization in different effector populations. Because of this ability, DCs have been used in recent decades to develop cancer immunotherapy approaches. In this type of cellular immunotherapy, the production of ex vivo DCs is necessary, and there are several protocols for this purpose. The aim of the present work was to compare two differentiation protocols, Fast (1 day) and Conventional (6 days), while four different maturation cocktails were also tested, where samples from 8 healthy donors were used. For this comparison, the immunophenotypic profile, uptake capacity, cytokine production and metabolic activity were assessed. The results showed that, despite some differences, the cells differentiated by the Fast protocol had a phenotypic profile and uptake capacity very similar to Conventional DCs. Regarding the response to the different maturation conditions, both Fast and Conventional CDs matured more effectively when stimulated with the Alpha cocktail or TLR ligands. At the metabolic level, the main differences between protocols were related to the higher glycolytic and/or glutaminolytic activity of the cells differentiated by the Conventional method. In terms of maturation, the Alpha cocktail appeared to stimulate glutaminolysis, while maturation with the Standard cocktail appeared to favor glycolysis. |
id |
RCAP_0e97bc2e3f0f003ce25e1140010f5dfe |
---|---|
oai_identifier_str |
oai:ria.ua.pt:10773/29894 |
network_acronym_str |
RCAP |
network_name_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository_id_str |
7160 |
spelling |
Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocolsMonocyte derived dendritic cellsEx vivo differentiation protocolFast vs. conventionalMaturation cocktailsMetabolism and metabolomicsDendritic cells (DCs) are cells specialized in antigen presentation that play an important role in the connection between innate and adaptive immunity. These cells have the unique ability to recognize, capture, process and present antigens to naïve T cells, leading to their polarization in different effector populations. Because of this ability, DCs have been used in recent decades to develop cancer immunotherapy approaches. In this type of cellular immunotherapy, the production of ex vivo DCs is necessary, and there are several protocols for this purpose. The aim of the present work was to compare two differentiation protocols, Fast (1 day) and Conventional (6 days), while four different maturation cocktails were also tested, where samples from 8 healthy donors were used. For this comparison, the immunophenotypic profile, uptake capacity, cytokine production and metabolic activity were assessed. The results showed that, despite some differences, the cells differentiated by the Fast protocol had a phenotypic profile and uptake capacity very similar to Conventional DCs. Regarding the response to the different maturation conditions, both Fast and Conventional CDs matured more effectively when stimulated with the Alpha cocktail or TLR ligands. At the metabolic level, the main differences between protocols were related to the higher glycolytic and/or glutaminolytic activity of the cells differentiated by the Conventional method. In terms of maturation, the Alpha cocktail appeared to stimulate glutaminolysis, while maturation with the Standard cocktail appeared to favor glycolysis.As células dendríticas (CDs) são células especializadas na apresentação antigénica que desempenham um importante papel na ligação entre a imunidade inata e a imunidade adaptativa. Estas células têm a capacidade única de reconhecer, capturar, processar e apresentar antigénios a células T naïve, levando à polarização destas nas suas diferentes populações efetoras. Devido a esta capacidade, as CDs têm sido usadas nas últimas décadas no desenvolvimento de abordagens de imunoterapia contra o cancro. Neste tipo de imunoterapia celular é necessária a produção das CDs ex vivo, existindo para tal vários protocolos. O objetivo do presente trabalho prendeu-se com a comparação de dois protocolos de diferenciação, Rápido (1 dia) e Convencional (6 dias), sendo também testados quatro cocktails de maturação diferentes, tendo sido utilizadas amostras de 8 dadores saudáveis. Para esta comparação, foram analisados o perfil imunofenotípico, a capacidade de uptake, a produção de citocinas e a atividade metabólica. Os resultados obtidos demonstraram que apesar de algumas diferenças, as células diferenciadas pelo protocolo Rápido apresentaram um perfil fenotípico e capacidade de uptake bastante semelhante às CDs Convencionais. Relativamente à resposta às diferentes condições de maturação, tanto as CDs Rápidas como as Convencionais maturaram de forma mais efetiva quando estimuladas com o cocktail Alpha ou ligandos de TLR. A nível metabólico, as principais diferenças entre os protocolos prenderam-se com a maior atividade glicolítica e/ou glutaminolítica das células diferenciadas pelo método Convencional. Em termos de maturação, o cocktail Alpha pareceu estimular a glutaminólise, enquanto que a maturação com o cocktail Standard pareceu favorecer a glicólise.2021-12-21T00:00:00Z2019-12-20T00:00:00Z2019-12-20info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10773/29894engMendes, Cátia da Silvainfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-22T11:57:51Zoai:ria.ua.pt:10773/29894Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:02:09.405797Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocols |
title |
Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocols |
spellingShingle |
Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocols Mendes, Cátia da Silva Monocyte derived dendritic cells Ex vivo differentiation protocol Fast vs. conventional Maturation cocktails Metabolism and metabolomics |
title_short |
Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocols |
title_full |
Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocols |
title_fullStr |
Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocols |
title_full_unstemmed |
Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocols |
title_sort |
Analysis of monocyte-derived dendritic cells obtained through conventional vs. fast differentiation/maturation protocols |
author |
Mendes, Cátia da Silva |
author_facet |
Mendes, Cátia da Silva |
author_role |
author |
dc.contributor.author.fl_str_mv |
Mendes, Cátia da Silva |
dc.subject.por.fl_str_mv |
Monocyte derived dendritic cells Ex vivo differentiation protocol Fast vs. conventional Maturation cocktails Metabolism and metabolomics |
topic |
Monocyte derived dendritic cells Ex vivo differentiation protocol Fast vs. conventional Maturation cocktails Metabolism and metabolomics |
description |
Dendritic cells (DCs) are cells specialized in antigen presentation that play an important role in the connection between innate and adaptive immunity. These cells have the unique ability to recognize, capture, process and present antigens to naïve T cells, leading to their polarization in different effector populations. Because of this ability, DCs have been used in recent decades to develop cancer immunotherapy approaches. In this type of cellular immunotherapy, the production of ex vivo DCs is necessary, and there are several protocols for this purpose. The aim of the present work was to compare two differentiation protocols, Fast (1 day) and Conventional (6 days), while four different maturation cocktails were also tested, where samples from 8 healthy donors were used. For this comparison, the immunophenotypic profile, uptake capacity, cytokine production and metabolic activity were assessed. The results showed that, despite some differences, the cells differentiated by the Fast protocol had a phenotypic profile and uptake capacity very similar to Conventional DCs. Regarding the response to the different maturation conditions, both Fast and Conventional CDs matured more effectively when stimulated with the Alpha cocktail or TLR ligands. At the metabolic level, the main differences between protocols were related to the higher glycolytic and/or glutaminolytic activity of the cells differentiated by the Conventional method. In terms of maturation, the Alpha cocktail appeared to stimulate glutaminolysis, while maturation with the Standard cocktail appeared to favor glycolysis. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-12-20T00:00:00Z 2019-12-20 2021-12-21T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10773/29894 |
url |
http://hdl.handle.net/10773/29894 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
_version_ |
1799137676969902080 |