4.4 Article

Evaluation of cholesterol content and impact on antigen exposure in the outer lipid bilayer of adult schistosomes

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PARASITOLOGY
卷 134, 期 -, 页码 1775-1783

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CAMBRIDGE UNIV PRESS
DOI: 10.1017/S0031182007003174

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Schistosoma mansoni; Schistosoma haematobium; cholesterol content; methyl-beta-cyclodextrin; surface membrane antigens

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Developing and adult Schistosoma mansoni and S. haematobium intact worms do not bind specific antibodies, likely because of structural and boichemical modifications of the order lipid bilayer. We have estimated the amount of cholesterol in the apical membrane of adult schistosomes via extraction with the membrane-imperable, cholesterol-binding drug methly-beta-cyclodextrin (MBCD), followed by filipin staining of the worms, and evolution of the amount of cholesterol released in the medium by a commercially available, enzymatic colorimetric assay. Positive correlations between amount of released cholesterol, MBCD concentrations, and worm number and age provided evidence for the sensitivity and valilidity of the newly developed method. Treatment with 40 mM MBCD for 2h at 37 degrees C led to total loss of cholesterol from the worm outer membrane, as assessed by filipin staining, and the released cholesterol values were used to estimate the amount of cholesterol per worm and per an approximate surface area unit. Additionally, total depletion of outer membrane cholesterol was associated with exposure of surface membrane antigens to specific antibody binding in 50% and 70% of S. haematobium and S. mansoni worms, respectively. These findings together suggest that cholesterol is an essential, but not the sole, factor in sequestion of surface membrane antigens in schistosomes.

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