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| Author(s): |
Maria Carolina Ferreira
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Faculdade de Ciências Médicas |
| Defense date: | 2009-02-02 |
| Examining board members: |
Ronei Luciano Mamoni;
Niels Olsen Saraiva Camara;
Wirla Maria da Silva Cunha Tamashiro
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| Advisor: | Ronei Luciano Mamoni |
| Abstract | |
Patients with paracoccidioidomycosis (PCM) present suppression of cellular immune response characterized by negative DTH to P. brasiliensis antigens, elevated apoptosis of lymphocytes, high expression of CTLA-4, and production of IL-10 and TGF-ß. Together these data point towards the involvement of regulatory T cells (CD4+CD25+ - Tregs) in PCM. The aims of this study were to verify whether and how Tregs are involved in this immunosuppression, through the analysis of the number and the phenotype, as well as, the functional activity of Tregs cells from peripheral blood of patients before and after antifungal treatment. The number and phenotype of Tregs cells were evaluated by flow cytometry, and the results showed that PCM patients with active disease (AD) present higher number of Treg cells than patients after treatment (TD) or healthy controls (C). Furthermore, we observed higher expression of CD95L, CTLA-4, LAP-1 and GITR on Tregs from AD group, than in cells from TD and C groups. mRNA expression (quantified by qRT-PCR) for FoxP3, IL-10 and TGF-ß in ex vivo PBMC, were also higher in AD group than in controls and in TD group. In order to compare the functional activity of Tregs, we analyzed the effect of Treg cells on the proliferative response of PBMC stimulated with ConA. Tregs from AD group exhibited stronger regulatory activity than cells from TD and C groups, in a dose dependent manner. To verify the possible mechanism through which Tregs suppress the cell proliferation we utilized a transwell system, which showed that the contact is mandatory for regulatory activity of Treg cells from C group, but had only a partial involvement in cells from AD patients. The addition of IL-10 and TGF-ß and anti cytokines in the co-cultures showed that the production of immunoregulatory cytokines may be other mechanism used by Tregs. In conclusion, the increased number of Treg cells in peripheral blood of patients with active disease, expressing high levels of regulatory markers and suppressive activity, besides the high expression of Foxp3, IL-10 and TGF-ß mRNAs suggest the potential participation of this population in the immunosuppression observed during the disease. Moreover, our results indicate that Tregs act mainly by contact to exert their inhibitory effects, but the production of immunoregulatory cytokines are also involved. (AU) | |