Proteomic Identification of IPSE/alpha-1 as a Major Hepatotoxin Secreted by Schistosoma mansoni Eggs
Figure 3
Omega (ω)-1 exhibits dose-dependent hepatotoxicity that is neutralized by a specific monoclonal antibody.
(A) Hepatocyte cultures (0.5 ml in Dulbecco's modified Eagle's medium were co-incubated with various amounts of purified native omega-1, and egg-derived material (10 µg/ml). After 72 h, ALT, a biomarker for hepatotoxicity, was measured. Polymyxin B (80 µg/ml) was included in co-incubations with omega-1 to neutralize any potential LPS. Control cultures contained polymyxin B alone. Aspergillus oryzae T2 RNAse (25 U/ml) was also used as a comparison in light of omega-1's described T2 RNase activity. A combination of 5 mM D-galactosamine hydrochloride (D-gal) and 1 µg/ml rTNF-α (D-gal/TNF-α) was employed as a known hepatotoxic control and negative control cultures used PBS. (B) Pre-incubation of a specific monoclonal antibody (5 µg/ml) with omega-1 abolished the latter's toxicity and respectively decreased cytotoxicity of ESP and SEA by 47 and 33%. Data are presented as the means ± SD from two independent experiments each performed in duplicate. *P<0.02, **P<0.001 and ***P<0.0001 using a one-sided paired Student's t-test.