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Doctoral Thesis
DOI
10.11606/T.42.2008.tde-18112008-164720
Document
Author
Full name
João Paulo Martins do Carmo
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Barbuto, Jose Alexandre Marzagao (President)
Câmara, Niels Olsen Saraiva
Kallas, Esper Georges
Negro, Sonia Jancar
Postól, Edilberto
Title in Portuguese
Modulação fenotípica e funcional de células dendríticas derivadas in vitro de monócitos por contato com linfócitos preé-aquecidos e/ou irradiados.
Keywords in Portuguese
Apoptose
Células dendríticas
Citocinas
Estresse por aquecimento
Radiação ionizante
Tolerância imunológica
Abstract in Portuguese
DCs são células especializadas na apresentação de antígenos (Ags) para linfócitos T virgens e indução de respostas imunes primárias. Deficiências no processo de eliminação de células apoptóticas relacionam-se com desenvolvimento de doenças auto-imunes. Para avaliar o efeito de células apoptóticas sobre os processos de maturação e atividade funcional de DCs derivadas in vitro de monócitos aderentes de doadores saudáveis, células não aderentes obtidas após uma 2ª etapa de aderência por 3 dias foram submetidas a aquecimento e/ou irradiação. 48h após, células somente irradiadas (37i) apresentaram maior porcentagem de apoptose que células aquecidas e irradiadas (43i), sugerindo que o calor protege da apoptose induzida. Na cocultura com iDCs ou mDCs (iDC+TNF), iDC+37i apresentaram aumento de CD1a, correlacionado com altos níveis de IL-10 e inibição de autoestimulação linfocitária. mDC+43i apresentaram níveis de CD1a semelhantes a iDC, baixos níveis de IL-10, altos níveis de IL-12p70 e altos índices de auto e aloestimulação linfocitária. Concluímos que o fenótipo e função de DCs é modulado diferencialmente na presença de 43i, que induzem ativação in vitro, enquanto 37i induzem DCs com características moduladoras dependente de lipídios.
Title in English
Phenotypic and functional modulation of human monocyte-derived dendritic cells after in vitro interaction with pre-heated and/or irradiated lymphocytes.
Keywords in English
Apoptosis
Cellular stress
Cytokines
Dentric cells
Immunological tolerance
Ionizing radiation
Abstract in English
DCs are specialized in presenting Ags to naïve lymphocytes, inducing primary immune responses or immunological peripheral tolerance, through tissue turnover by scavenging dying cells. Works with human cells are scarcer and controversial about the latter. Then, the aim of this work was to investigate the effect of apoptotic cells (Nadhs) on maturation and function of human monocyte-derived DCs in vitro. The results suggest that heating Nadhs before irradiation (43i) seems to protect them from apoptosis induced by irradiation (37i). DCs were cocultured with 37i or 43i, simultaneously to TNF-a addition in the 5th day of culture. In the 7th day, there was an association between high levels of CD1a, IL-10, low levels of IL-12p70 and decreased allostimulation induced by iDC+37i. mDC+43i had high levels of IL-12p70, CD86 and proliferation index, associated with low levels in CD1a expression. We conclude that DCs phenotype and function are differentially modulated in the presence of 43i, which induce DCs activation, or 37i, which induce regulatory phenotype and function in DCs. We suggest that these protocols for DCs activation with 43i or 37i could be used, respectively, as models of in vitro auto-reactivity or homeostatic immunomodulation of DCs in vitro.
 
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Publishing Date
2009-02-18
 
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