Isolation & immunochemical characterization of diagnostically relevant antigens of Echinococcus granulosus.
Two hydatid specific polypeptides with molecular masses of 8 kDa and 116 kDa have been successfully isolated from E. granulosus hydatid cyst fluid using affinity chromatography. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and western immunoblot analysis under reducing and denaturing conditions indicated the 116 kDa purified antigen to be a hetero-tetramer consisting of 45 kDa, 66 kDa, 75 kDa and 116 kDa subunits linked by disulphide bonds while the 8 kDa purified antigen was found to be a monomer polypeptide. Affinity purified 116 kDa molecule was heat-labile, sensitive to treatment with pronase, trypsin and pepsin and its immunoreactivity as assessed in enzyme linked immunosorbent assay remained unaltered on treatment with sodium metaperiodate. The affinity purified 8 kDa molecule was heat-stable, sensitive to proteolytic enzymes and also sodium metaperiodate oxidation. Lectin binding studies revealed that the 8 kDa molecule specifically bound Concanavalin A and Triticum vulgaris, and thus had varies; is directly proportional to-D-glucose and N-acetyl D-glucosamine sugar moieties. The immunoreactivity of both the antigens remained unaltered on treatment with lipase. However, biochemical estimation of total lipid content revealed the affinity purified 116 kDa antigen to contain 6.25 per cent total lipids suggesting it to be lipoproteinic in nature. The 8 kDa antigen had no detectable total lipids biochemically. All sera from patients confirmed to have hydatidosis recognised the 8 kDa and 116 kDa polypeptides. However, sera from seven subjects with other parasitic infections also recognised the 116 kDa antigen though not the 8 kDa antigen. The data suggested that the recognition of 8 kDa antigen of E. granulosus has potential for specific immunodiagnosis of hydatidosis.
Department of Experimental Medicine, Postgraduate Institute of Medical Education & Research, Chandigarh.
J R
JR
Kanwar
V K
VK
Vinayak
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D004443:Echinococcosis / Q000175:diagnosis* / Q000276:immunology; D004446:Echinococcus / Q000276:immunology*; D006801:Humans; D008970:Molecular Weight; D011817:Rabbits; D012756:Sheep
D000909:Antibodies, Helminth; D000947:Antigens, Helminth