Antigenic polypeptides of Echinococcus granulosus oncospheres and definition of protective molecules.
Immunoblotting and in vitro oncosphere-killing were used to identify a putative protective molecule in Echinococcus granulosus mature oncospheres. A range of sera from sheep that had been shown to be protected against E. granulosus, and from those that were not, were tested. The sera used were obtained from sheep hyperimmunized with E. granulosus oncospheres, or immunized with oncosphere non-denatured extract, with immature oncosphere extract or with denatured extracts of oncospheres. Results indicated the involvement of native antigens of 23, 25, 30, 34 and 40 kDa in the protective response to E. granulosus infection. The rapid appearance of antibodies to the 23, 25 kDa antigens, their association with early onset of protection and in vitro oncosphere lysis by affinity-purified antibodies obtained from these fractions, indicated that these antigens contained protective epitopes. Final confirmation was provided by immunization of sheep with fractions prepared by preparative SDS/PAGE, and challenge infection. Only the fraction containing the 23 and 25 kDa molecules was able to stimulate protection. Antisera against this pair of molecules should provide a useful probe for screening an E. granulosus oncosphere cDNA library to identify clones expressing protective molecules.
Heath
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DD
AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand.
Lawrence
S B
SB
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000737:chemistry / Q000302:isolation & purification*; D003602:Cytotoxicity, Immunologic; D004443:Echinococcosis / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary; D004446:Echinococcus / Q000276:immunology*; D015801:Helminth Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D007114:Immunization; D015151:Immunoblotting; D066298:In Vitro Techniques; D008970:Molecular Weight; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology / Q000517:prevention & control
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins