Immunochemical characterization of alkaline phosphatase from the fluid of sterile and fertile Echinococcus granulosus cysts

Abstract

Alkaline phosphatase (ALP) from hydatid cyst fluid (HCF) was purified and characterized for comparison between fertile and sterile HCF. Samples were obtained from slaughtered sheep and then sterile and fertile cysts were separated. ALP was purified from aspirated cyst fluid and biochemical parameters were determined. Sera from patients with hydatid disease (15 samples) and patients with other parasitic diseases including fascioliasis (2 samples), taeniasis ( Taenia saginata, 5 samples) and also sera from uninfected controls (15 samples), were collected and used in immunoblotting experiments with ALP from sterile and fertile HCF as antigen. Our results showed that ALP activity in fertile HCF [10.75+/-3.78 (SD) U/ml) was significantly more than in sterile HCF (6.25+/-2.43 U/ml). There were also some differences between the kinetic parameters and biochemical characteristics of ALP in fertile and sterile HCF. Immunoreactive bands were clearly observed when sera from hydatid infected patients were tested with ALP from fertile HCF as the antigen. However, this method revealed no cross-reaction between purified ALP from sterile HCF and sheep liver tissue. These findings suggest that there is some variation in the immunochemical characteristics of ALP from fertile and sterile HCF.

Apoptosis as a possible mechanism of infertility in Echinococcus granulosus hydatid cysts

“…Two types of hydatid cysts can be observed in intermediate hosts: fertile cysts, on which protoscoleces are both joined to the germinal layer and free in the hydatid fluid filling the cyst cavity, and infertile cysts, which do not produce protoscoleces and are therefore unable to continue with the life cycle of the parasite [Kamenetzky et al, 2000; Lahmar et al, 2004]. The biological and molecular background underlying the generation of both types of cysts have not yet been elucidated [Vatankhah et al, 2003].…”

Characterization of cellular and humoral immune responses to alkaline phosphatase from fertile hydatid cysts in the human peripheral blood

“…Zhang et al, 2017). In our previous study, we also showed significant differences between fertile and sterile cysts according to the kinetics and immunochemical characteristics of E.ALP (Vatankhah, Assmar, Vatankhah, & Shokrgozar, 2003).…”

“…The enzyme was also proposed to be a marker for analyzing the effect of chemotherapeutical and interventional treatments of hydatid infection (Adnani Sadati, Farahnak, Molaei Rad, Golestani, & Eshraghiyan, ; Feng et al, ; R. Zhang et al, ). In our previous study, we also showed significant differences between fertile and sterile cysts according to the kinetics and immunochemical characteristics of E.ALP (Vatankhah, Assmar, Vatankhah, & Shokrgozar, ). Besides, detectable levels of antibodies against this enzyme in the sera of human patients confirmed that E.ALP can activate specific immune reactions in the host (Mahmoud & Abou Gamra, ).…”

“…E.ALP was also purified from the extracts of 30 fertile cysts, according to the method described elsewhere (Vatankhah et al, 2003).…”

“…Protein extracts were fractionated by SDS-PAGE; the procedure was followed by blotting the electrophoretic bands on nitrocellulose membranes that had been incubated overnight at 4°C in 1:64 dilution of each serum sample from patients with CE and normal donors, as it was previously explained (Vatankhah et al, 2003).…”