Purified laminar layer and hydatid fluid of Echinococcus granulosus modulates dendritic cells improving global translationby activating mTOR pathways and promotes cell maturation and T cell proliferation.

NICOLAO, MARÍA CELESTE; CUMINO, ANDREA C.; RODRIGUEZ RODRIGUES, CHRISTIAN

Lima

Congreso; XXVIII World Congress on Echinococcosis; 2019

The World Association of Echinococcosis (WAE), the Peruvian Association of Hydatidology (PAH), Universidad Nacional Mayor de San Marcos and Universidad Peruana Cayetano Heredia

Introduction: The target of rapamycin (mTOR), represents a biological switch in themodulation of cell metabolism and environmental signals. It has recently been recognized as aregulator in the immune system. The aim of this workis to analyze if Echinococcus antigenscould trigger an immune response by activating mTOR pathway in dendritic cells. Methods:BMDCs were culturedin complete RPMI. Hydatid fluid (HF) was punctured from the hydatidcysts collected of infected cattle slaughtered. The germinal layer was removed from purifiedlaminar layers (pLL) by washing with 2M NaCl. Cytotoxicity, phagocytosis, maturation ofBMDCs and T cell proliferation were analyzed by FACS (mAbs directed to CD11c, CD40,CD80, CD86, MHC I and MHC II were used). Translation and phospho-mTOR were evaluatedby Immunoblotting and confocal microscopy. Results: First, we evaluate cytotoxicity. Neitherof the two antigens induce loss of cell viability. Protein synthesis was monitored usingpuromycin labeling. Cells not treated with puromycin, cycloheximide or rapamycin were usedas controls. Both stimulus, pLL (MFI: 979) and HF (MFI:835), showed an increase intranslation levels compared to its counterpart without stimulation (MFI:686, n=3). Furthermore,we have observed that DCs cultured without SFB or FLT3L, and stimulated with pLL or HFinduce an increase in mTOR phosphorylation levels compared to untreated CDs (n=3, p=0,035Anova test). Next, we evaluated the phenotype of DCs, both antigens downregulatephagocytosis levels (CTRL:80% vs PLL:75% or HF:66% n=3) and pLL of induce an upregulation of MHCII and CD86 (n=3, p