Bovine - Echinococcus granulosus cell line (EGPE) as antigenic support for hydatidosis diagnosis and follow up

MAGLIOCO A; MELISA S BARBERY VENTURI; JORGE GENTILE; CLAUDIA HERNANDEZ; OSCAR JENSEN; MARÍA LAURA GERTISER; ALICIA G FUCHS

Mar del Plata

Congreso; LXIV Reunión Anual de SAI; 2016

 Introduction: Echinococcus granulosus is thecausative agent of hydatid disease (HD), a widely distributed zoonosis in theworld. The usual source of antigens used for immunodiagnosis is the hydatid cystfluid. Differences were observed in the specificity and sensitivity betweendifferent studies, perhaps due to the use of a source of non-standardizedantigens. In our laboratory, a cell line from bovine protoscolices (EGPE cells)has been established (patent first instance approved INPI P-090102320). Inthese cells, the antigen B was detected by immunohistochemistry and DCO1 wasdetected by PCR (Echeverria et al 2010). Aim:to validate the use of antigens from EGPE cells, as a standardized sourceof antigens, to diagnose HD and monitoring its follow up.  Studydesign: Presence of relevantantigens to HD were evaluated in EGPE, by western blotting. Three differentprotein mixtures were assayed: proteic extract from short or long culture ofEGPE and supernatant. Serum from 13 patients with HD or non HD patients from anendemic zone (Tandil) were used in a case-control study. Six of these HDpatients were evaluated 4 years later. Bands detected exclusively by HDpatients were taking into account as positive band. Results: The combination of different antigens from the 3 sourcesallow the diagnosis of 13/13 patients. The three sources have different proteincomposition. The bands detected by HD patients in short culture antigens were(KDa): 76-78, 70-73, 64-65, 58-60, 52-55, 44-47 and 40-41; in long cultureantigens detected were (KDa): 93-94, 78-81, 73-77, 69-72, 63-66, 56-57, 51-55,46-47 and 40-42; and in supernatant antigens detected were (KDa): 86-90, 79-80,75-76 and 58-60. These bands were lost in 5/6 HD patients 4 years later(p<0.05, Chi-square test). Conclusion:More than one antigen is necessary to HD diagnosis and EGPE proteins would be avery useful source of antigens for HD diagnosis and follow up. This study was approved by UAI ethics committee.