Demonstration of anti-cuticular antibodies by immuno-electron microscopy in sera of mice immunized with cuticular extracts and isolated cuticles of adult Dipetalonema viteae (Filarioidea).
The immunogold technique was used for the ultrastructural localization of antibody-binding sites on thin sections of Lowicryl K4M embedded adult females, infective larvae and pieces of adult cuticles of Dipetalonema viteae insoluble in SDS-2-ME. The antisera used were either produced against SDS-2-ME extracts of cuticles or the insoluble pellet after SDS-2-ME extraction. With both types of antisera a labelling of epitopes on fibers was achieved in intact cuticles. In isolated cuticles the corresponding structures were absent. The same sera crossreacted with the larval and microfilarial cuticle as well as with somatic structures of all three stages. The only serum against isolated cuticle, which did not recognize cuticle fibers also did not crossreact with somatic structures. The recognition of the electron dense cortical layer insoluble in SDS-2-ME depended on the number of immunizations. A labelling of the filarial surface was never achieved. A dense labelling of the apical membrane enfoldings of the hypodermis pointed to an involvement in the synthesis of the nematode cuticle. The rough endoplasmic reticulum in the apex of the epithelial cells of the uterus, the content of the nutrient channels, and the substance between eggshell and microfilariae crossreacted with most of the antisera. This led to the conclusion that these substances are partly produced by the uterus epithelium.
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D003429:Cross Reactions; D004153:Dipetalonema / Q000276:immunology* / Q000648:ultrastructure; D000939:Epitopes / Q000276:immunology; D005260:Female; D005455:Fluorescent Antibody Technique; D006046:Gold; D006651:Histocytochemistry; D007114:Immunization; D007158:Immunologic Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron / Q000379:methods
D000906:Antibodies; D000939:Epitopes; D006046:Gold