Vaccination against hepatic amebiasis in hamsters.
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Biomedical subjects
Publications and source records attributed to E Meerovitch.
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The lost pathogenicity of two strains of Entamoeba histolytica, one isolated in 1924 and the other in 1967, grown in axenic culture for the past five and six years respectively, was restored by supplementing the culture medium with cholesterol through a number of transfers. The number of passages in the cholesterol supplemented medium, necessary to restore a certain degree of pathogenicity of the two strains in hamsters, was proportional to the total time of in vitro cultivation of the strain, and not just the time of cultivation under axenic conditions. Pathogenicity, once restored, persisted for a long time after cholesterol treatment was stopped.
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A cell fractionation scheme was developed for studying the distribution of certain hydrolases, especially phosphohydrolases in a Crithidia sp. (Trypanosomatidae). Whilst between 26-56% of the total cellular hydrolase activities were soluble (probably of flagellar pocket origin), a certain percentage, 5-40%, was sedimentable. A particulate fraction obtained after isopycnic density gradient centrifugation (p = 1.187-1.241), designated fraction FA/FB, was enriched in various acid hydrolases (relative specific activities 1.33-6.24) and displayed latent phosphohydrolase activities. The density gradient distributions of this hydrolytic enzymes were compared with reference to one another and malate dehydrogenase (mitochondrial marker). From the results obtained it appears that the sedimentable acid hydrolases of Crithidia are associated with a heterogeneous population of subcellular particles. Cytochemical observations on the FA/FB fraction supported this finding and revealed the association of acid phosphatase reaction product with subcellular elements resembling multivesicular bodies.
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The antigenic properties of cell fractions prepared from axenically grown Entamoeba invadens were investigated using various immunoelectrophoretic methods. All membrane fractions displayed varying degrees of antigenicity, the most predominant being a cytoplasmic light-vesicle fraction. The surface membrane showed the least diversified range of antigenic components and was also the least immunogenic fraction as judged by the reactivity of the antisera produced. Using tandem-crossed immunoelectrophoresis the antigenic profiles of the membrane fractions were compared. No evidence was obtained for the presence of either lipid or carbohydrate in the antigenic moieties of any of the membrane fractions. Using a series of sequential solvent extractions it was concluded that both divalent metallochelate linkages and interiorly located hydrophobic associations were principally involved in binding the major antigenic components within the light-vesicle membrane. An enzymic function was assigned to certain of the membrane antigens, the immunoprecipitates obtained showing both acid phosphohydrolase and non-specific esterase acitivity toward a variety of substrates.
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