Identification and quantification of proteins involved in cell differentiation processes in germinal cells of Echinococcus granulosus.

ALBANI CLARA MARÍA; REINDERS JOERG; FABBRI JULIA; PENSEL, PATRICIA EUGENIA; ELISSONDO MARIA CELINA

Lima

Congreso; 28th World Congress on Echinococcosis; 2019

Cystic echinococcosis is a severe neglected zoonotic infection caused by the larval stage (metacestode) of the parasite Echinococcus granulosus (Eg), which has a worldwide distribution and important medical and economic impact. The inner layer of the hydatid cyst or germinal layer contains a group of cells called germinative cells which are responsible for cell proliferation and from which differentiated cell types develop. Using a proteomic strategy, we identify and quantify proteins involved in processes of cellular differentiation in Eg germinative cells grown in different conditions.Eg primary cell culture was obtained from hydatid cysts and maintained with weekly splitting during 1-4 month. Cells were culture in normal media and conditioned media (resembling host environment) for different time periods. Then, 50 µl of cells were homogenized in lysis buffer and the peptide content was estimated. Aliquots of 100 µg protein per sample were used for filter-aided sample preparation and the resulting peptides were subjected to nano-LC-MS/MS-analysis. A combined library was set-up by combining the different runs using the Protein Pilot-software and Uniprot database. We identified a total of 455 proteins in all the conditions studied. We identified and quantified 9 proteins associated with cell differentiation which were differentially represented in the conditions studied. For example HSP 90 alfa (Accession W6UM50) and Neurogenic notch (Accession W6UG53) were found overrepresented in cells cultured in normal media with a fold change 3,2 and 4,8 respectively (p < 0,05) in comparison with cells culture in conditioned media (resembling host environment).Through the methodology used, it was possible to determine several proteins associated with cell differentiation present in the germinative cells of Eg and to hypothesize its possible function. Moreover many of the identified proteins could be useful as cestode developmental markers.