Echinococcus granulosus promotes autophagosome nucleation and LC3-attachment to the autophagic membrane in dendritic cells.

CHOP, MAIA; LOOS JULIA A; NICOLAO MA. CELESTE; CUMINO ANDREA C.; RODRIGUEZ R CHRISTIAN

Bristol

Workshop; Autophagy UK Meeting; 2021

Lane & Carroll UK Meeting

Background: Autophagy is a cellular catabolic process and plays a key role in the presentation of endogenous and exogenous proteins on MHC I and II molecules, promoting activation of CD8+ and CD4+ T cells respectively. The aim of this work is to analyze if hydatid fluid (HF) from Echinococcus, constituted by a wide range of parasite proteins could trigger autophagy in dendritic cells. Methods: BMDCs were cultured in complete RPMI. Hydatid fluid (HF) was punctured from the hydatid cysts collected of infected cattle slaughtered. Autophagy induction was evaluated by Transmission Electron Microscopy (TEM) and immunofluorescence confocal microscopy with Rabbit anti LC3-antibody clone H50. Autophagy-related genes were analyzed by quantitative PCR and autophagy flux was analyzed by the use of Chloroquine (100 M 4 h), an inhibitor of acidification of lysosomes. Results: We have evaluated changes in the basal level of autophagy on BMDCs upon HF stimulation. HF-stimulated BMDCs significantly enhanced the mean fluorescence intensity of MHC II and LC3 and showed a trend in the increment of number and the average size of LC3-positive structures in comparison with unstimulated cells (*p