Identification of a laminated layer-associated protein in Echinococcus multilocularis metacestodes.
Echinococcus multilocularis is a cestode parasite that predominantly infects red and arctic foxes as definitive hosts. Ingestion of E. multilocularis eggs and subsequent post-oncospheral infection with the larval stage (metacestode) of the parasite results in alveolar echinococcosis (AE), a life-threatening hepatic disease concerning humans and other intermediate hosts such as small rodents. The primary fluid-filled vesicles of the asexually proliferating metacestode are comprised of an inner germinal layer, a syncytial tegument, and an outer, acellular, so-called laminated layer. This laminated layer may play an important role in protecting the developing E. multilocularis metacestode from host immune reactions, and laminated layer-associated components represent potential targets for intervention during the course of AE. We have used an in vitro cultivation technique for the long-term maintenance and proliferation of E. multilocularis metacestodes in order to generate premature (protoscolex-free) parasite vesicles. A polyclonal antiserum was raised against this host-free parasite tissue. Subsequent immunoblot analysis of parasite fractions obtained by Triton X-114 extraction lead to the identification of a 116 kDa protein (named EmP2) within the Triton-insoluble fraction. The characterization of EmP2 by SDS-PAGE, Western blotting, and by immunofluorescence revealed that EmP2 is a laminated layer-associated protein.
Ingold
K
K
Institute of Parasitology, University of Berne, Switzerland.
Gottstein
B
B
Hemphill
A
A
D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology / Q000302:isolation & purification; D000915:Antibody Affinity; D000947:Antigens, Helminth / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D004443:Echinococcosis / Q000469:parasitology; D004446:Echinococcus / Q000254:growth & development / Q000276:immunology*; D019084:Fluorescent Antibody Technique, Indirect; D015801:Helminth Proteins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D006790:Host-Parasite Interactions; D007150:Immunohistochemistry; D007814:Larva / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D017830:Octoxynol; D011092:Polyethylene Glycols
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40