CONICET | Buscador de Institutos y Recursos Humanos

The target of rapamycin (mTOR), represents a key biologicalswitch in the modulation of cell metabolism and environmentalsignals. It has recently been recognized as a regulator in the immunesystem. In the present work, we show the modulation exertedby Echinococcus on signaling of the mTOR pathway and its relationto the functionality of dendritic cells. Methods: BMDCs were culturedin complete RPMI supplemented with 100 ng/ml Flt3-L. Hydatidcysts were collected from the liver of infected cattle slaughtered.Hydatid fluid (HF) was punctured from the cysts. The germinal layerwas removed from purified laminar layers (pLL) by washing with2M NaCl. Flow cytometry: FITC or PE-conjugated mAbs directed toCD11c, CD40, CD80, CD86, MHC I and MHC II were from eBioscience.Immunobloting and confocal microscopy: mouse anti-puromycinand rabbit anti-Phospho-mTOR antibodies were used. Results:Protein synthesis was monitored using puromycin labeling followedby confocal microscopy. Cells not treated with puromycin, cycloheximideor rapamycin were used as controls. Both stimulus, pLL(MFI:_979) and HF (MFI:_835), showed an increase in translationlevels compared to its counterpart without stimulation (MFI:_686,n=3). LPS was used as a positive control. To confirm if the mTORpathway was involved in the recognition of parasitic antigens, wemonitored the phosphorylation of MTORC1 by WB and confocalmicroscopy. We have observed that DCs cultured without SFB orFLT3L, and stimulated with pLL or HF induce an increase in mTORphosphorylation levels compared to untreated CDs (n=3, p=0,035Anova test). Finally, we evaluated the phenotype of DCs. The pLLof Echinococcus induce an up-regulation of MHCII and CD86 (n=3,p

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