Immunohistochemical localization of Gnathostoma spinigerum larval antigens by monoclonal antibodies: II. Electron microscopy

Abstract

Immunocytochemical localization of antigens in advanced third-stage larvae of Gnathostoma spinigerum (GsAL3) was studied by immunogold labeling method using seven G. spinigerum specific monoclonal antibodies (MAbs), FS-3D11, SS-5H5, SK-6C4, SK-4E1, SK-7G6, SK-8D4 and SA-9B5. All these MAbs belong to the IgG1subclass and only FS-3D11 and SS-5H5 recognize carbohydrate epitopes. The paraformaldehyde-fixed GsAL3 were embedded in Lowicryl K4M medium, and the gold colloidal particles used were 15 nm in size. When the worm sections were probed with FS-3D11, the gold particles appeared to concentrate specifically on the intestinal brush border. When SS-5H5 was applied, the particles were scattered densely over the brush border and in the cytoplasm of epithelial cells. The rest of the MAbs which recognize protein determinants exhibited a lack of labeling. The results suggested that the carbohydrate antigenic determinants recognized by the two MAbs are the most stable and most abundant particularly in the intestine of GsAL3. These results also confirmed the previous finding that the most antigenic site of GsAL3 is the intestine.