EGPE cells, analytical laboratory source for echinococcus granulosus antigens.

ANDREA MAGLIOCO; AGUERO F; ALEJANDRA JUÁREZ VALDEZ; JORGE GENTILE; CLAUDIA HERNANDEZ; OSCAR JENSEN; GERTISER, MARÍA LAURA; SÁNCHEZ ROMANÍ; CANZIANI, GABRIELA; FUCHS A

Exposici�n; Reunión anual de sociedades de biociencia; 2019

Cystic echinococcosis (CE) is a zoonoses worldwide distributed produced by Echinococcus granulosus sensu lato. In Argentina, CE is an endemic disease with active dissemination reporting more than 450 human cases per year. Disease diagnosis is performed by ultrasound. Serology tests for diagnosis, population screening and patients follow-up have poor or variable sensitivity and specificity. A cell line from E. granulosus G1 (EGPE cells) obtained in our laboratory (Echeverría et al, 2010) expresses antigen B and protein extracts from EGPE in different culture stages were recognized by sera from CE patients with high sensitivity by Western blot. The aim of this study was to identify, by proteomics, the CE antigens present in EGPE from 20-day-old culture. We tested sera from 34 CE patients (Tandil, Chubut and Perú), 21 healthy donors (Tandil), 5 cysticercosis (Perú) and 3 fascioliasis (Perú). Protocols were approved by the UAI Ethical Committee. Protein extracts were obtained with ice-cold lysis buffer, after 20 days of cell culture. Sera reactivity was detected with AP- goat anti-human IgG and BCIP/NBT, bands were analyzed with GelAnalyzer software. Protein extracts were separated through Sephacryl S-200 and further by affinity column performed with a pool of antibodies from: CE patients sera (CE column) or sera from patients with other parasitoses (OP column). Eluted proteins from affinity columns were identified by proteomics (CEQUIBIEM - FCEyN, UBA). CE patients sera recognized bands from 12 to 94 kDa, few bands were also recognized by sera from healthy donors or from patients with other parasitoses. Elution profile of the gel filtration column showed three peaks, all of them recognized by CE patients´ sera. Proteins obtained from CE column allowed the identification of 15 proteins from E. granulosus. No detectable proteins were identified from OP column. EGPE cells can be used as a laboratory tool for identification of epitopes involved in the immune response.