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Master's Dissertation
Full name
Márcio José Ferreira
Knowledge Area
Date of Defense
São Paulo, 2008
Bevilacqua, Estela Maris Andrade Forell (President)
Daher, Silvia
Oliveira, Sergio Ferreira de
Title in Portuguese
Expressão de IFN-gama e interleucina (IL)-10 e seus receptores pelas células trofoblásticas de camundongos.
Keywords in Portuguese
Interferon tipo II
Interleucina 10
Abstract in Portuguese
Analisamos a expressão de IL-10, IFN-g e, seus receptores pelas células trofoblásticas de camundongos, citocinas anti e pró-inflamatórias. Cones ectoplacentários aos 7,5 dias de gestação foram cultivados por 48 h e em seguida tratados com 100 U/mL IFN-g ou 10 hg/mL IL-10. Após 6 h e 14 h, as amostras foram processadas para análise da expressão gênica por RT-PCR e protéica por imunohistoquímica, respectivamente. Grupos controle não receberam tratamento. IFN-g aumentou a expressão de IL-10R1 mas não a de IL-10 nas células trofoblásticas. IL-10, ao contrário, aumentou a expressão de IFN-g, mas diminuiu IFN-gRa e não alterou IFN-gRb. Reações imunohistoquímicas confirmaram os resultados de expressão gênica. Isto sugere que o trofoblasto pode participar da imunidade da interface materno-placentária aumentando a expressão de IFN-g em situações em que no meio há aumento de citocinas anti-inflamatórias, o que deve ser o reflexo da necessidade e importância desta citocina para o sucesso da gestação.
Title in English
Expression of IFN-gamma and interleukin (IL)-10 and its receptores in the mouse trophoblast cells.
Keywords in English
Interferon II
Interleukin 10
Abstract in English
Key cytokines such as IL-10 and IFN-g, essential for immune response regulation, have also been found locally at maternal-placental interface during mouse pregnancy. Particularly, levels of IL-10 characterize an anti-inflammatory environment associated to the inhibition of T helper-1 lymphocytes (Th1) development and the proliferative stimulation of the B lymphocytes (humoral response). On the other hand, increases in IFN-g profile prevent T helper-2 lymphocytes (Th2) activation leading to an inflammatory response that favors a Th1 response. The local production of these cytokines by NK uterine cells and T gd lymphocytes are relevant, but not exclusive. Thus, this study analyzed the potential contribution of the trophoblast in the maintenance of Th1/Th2 balance in the maternal-placental interface, represented, respectively by the expression of IL-10 and IFN-g cytokines. The expression of the anti-inflammatory cytokine IL-10 and its receptor was evaluated in the presence of an inflammatory environment mimetized by IFN-g addition to the culture medium. On contrary, the expression of the IFN-g (and its receptor) was evaluated in the presence of IL-10 characterizing an anti-inflammatory condition. Mouse trophoblast cells were isolated from implantation sites at gestational day 7.5 and cultured in standard conditions. Gene and protein expression were determined by immunohistochemistry and RT-PCR. IL-10 and IFN-g and their receptors were expressed in cultured trophoblast cells in the absence or presence of IFN-g and IL-10, respectively. IFN-g treatment increased IL-10R1 expression but do not alter IL-10 expression. On contrary, in the presence of IL-10 the IFN-g expression increased significantly while the expression of its receptor decreased. These results suggest that a proinflammatory environment increases trophoblast responsiveness to IL-10 whereas an anti-inflammatory condition seems to reinforce the importance of IFN-g expression at the maternal-placental interface, on the initial periods of gestation.
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