Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2008.tde-03062008-110226
Document
Author
Full name
Márcio José Ferreira
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
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
Camundongos
Citocinas
Interferon tipo II
Interleucina 10
Receptores
Trofoblasto
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
Citokines
Interferon II
Interleukin 10
Mice
Receptores
Trophoblast
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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Publishing Date
2008-06-10