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Antigenic localities in the tissues of Paragonimus westermani by developmental stages using immunogold labeling method
H J Rim,S J Kim,*I J Sun and J S Lee
Department of Parasitology and Institute for Tropical Endemic Disease, College of Medicine, Korea University, Seoul 136-701, Korea.
*Department of Biology, Hallym University, Chunchon 200-702, Korea.
Abstract
In order to observe the antigenic localization in the tissues of Paragonimus westermani of developmental stages, immunogold labeling method was applied using serum of the cats which were infected with isolated metacercariae from Cambaroides similis. The sectioned worm tissues from each developmental stage were embedded in Lowicryl HM 20 medium, stained with infected serum IgG and protein A gold complex (particle size: 12 nm) and observed by electron microscopy. In the young adult worm tissue of 4 weeks after infection with metacercariae, the gold particles were specifically concentrated on the tegumental syncytium and cytoplasm of the tegumental cells as well as the secretory granules in the parenchymal tissue. The antigenic materials in the adult worm tissue were specifically concentrated on the secretory granules in the parenchymal tissue, the cytoplasm between granules in the vitelline gland and the epithelial lamella in the lumen of the caecum.
Figures
Figs. 1-5 Electron micrographs of the tegument of the Paragonimus westermani.
Fig. 1. The tegument of all developmental stages of the worm, which was reacted with cat sera from noninfected control, showed the tegumental syncytium (TS), basal layer (BL), circular muscle (CM) layer, interstitial matrix (IM) and tegumental cell cytoplasm (TCC). The gold particles were not labeled on the tegument or other portions of the tissue. Bar=1 µg (×13,000)
Fig. 2. The tegument of the worm reacted with IgG from cats at 3 weeks after infection. Gold particles were not labeled on the tegumental syncytium. Bar=1 µg (×25,000)
Fig. 3. The tegument of the worm at 4 weeks after infection reacted with IgG from cats at 12 weeks after infection. Gold particles were specifically labeled in the tegumental syncytium (TS). Bar=1 µg (×25,000)
Fig. 4. The tegument of the worm at 8 weeks of development reacted with IgG from cats at 12 weeks after infection. Gold particles were predominantly labeled in the tegumental syncytium (TS). Bar=1 µg (×25,000)
Fig. 5. The tegument of the worm at 12 weeks of development reacted with IgG from cats at 12 weeks after infection. Gold particles were slightly labeled in the tegumental syncytium (TS). Bar=1 µg (×25,000)
Figs. 6-9 Electron micrographs of the secretory granules in the parenchymal cell at all developmental stages of P. westermani.
Figs. 6&9. The parenchymal cells of the worm which reacted with IgG from cats at 3 weeks after infection showed well developed rough endoplasmic reticulum (RER) in the cytoplasm. Gold particles were slightly labeled on the secretory granules (SG). Bar=1 µg (×25,000)
Fig. 7. The parenchymal cell of the worm which reacted with IgG from cats at 8 weeks after infection. Gold particles were specifically labeled on the secretory granules. Bar=1 µg (×25,000)
Fig. 8. The parenchymal cell of the worm reacted which sera from cats at 12 weeks after infection. Gold particles were very specifically labeled on the secretory granules. Bar=1 µg ( ×25,000)
Figs. 10-13 Electron micrographs of the granules in the vitelline gland of the worm.
Fig. 10. The vitelline gland of the worm at 12 weeks of development reacted with IgG from cats at 3 weeks after infection. Gold particles were slightly labeled in the cytoplasm between granules (G). The parenchymal cell of the worm which reacted with IgG from cats at 8 weeks after infection. Gold particles were specifically labeled on the secretory granules. Bar=1 µg (×25,000)
Fig. 11. The vitelline gland of the worm at 12 weeks of development reacted with sera from cats at 12 weeks after infection. Gold particles were very specifically labeled in the cytoplasm between granules. Bar=1 µg (×25,000)
Fig. 12. The vitelline gland of the worm at 20 weeks of development reacted with IgG from cats at 12 weeks after infection. Gold particles were very specifically labeled in the cytoplasm between granules. Bar=1 µg (×25,000)
Fig. 13. The vitelline gland of the worm at 20 weeks of development reacted with IgG from cats at 4 weeks after infection. Gold particles were predominantly labeled in the cytoplasm between granules. Bar=1 µg (×25,000)
Figs. 14-18 Electron micrographs of the epithelial lamellae and lumen area in the caeca of the worm.
Fig. 14. The caeca of the worm which reacted with IgG from noninfected control showed well developed lamellae and lumen. Gold particles were not labeled on the lamellae and lumen matrix. Bar=1 µg (×25,000)
Fig. 15. The caeca of the worm which reacted with IgG from cats at 3 weeks after infection. Gold particles were slightly labeled on the lamellae and lumen matrix. Bar=1 µg (×25,000)
Fig. 16. The caeca of the worm which reacted with IgG from cats at 4 weeks after infection. Gold particles were predominantly labeled on the lamellae and lumen matrix. Bar=1 µg (×25,000)
Fig. 17. The caeca of the worm which reacted with IgG from cats at 8 weeks after infection. Gold particles were predominantly labeled on the lamellae and lumen matrix. Bar=1 µg (×25,000)
Fig. 18. The caeca of the worm which reacted with IgG from cats at 12 weeks after infection. Gold particles were specifically labeled on the lumen matrix. Bar=1 µg (×25,000)
Figs. 19-23 Electron micrographs of the excretory bladder of the worm at all developmental stages.
Fig. 19. The excretory bladder of the worm which reacted with IgG from noninfected control showed well developed lamellae in the excretory bladder (ELEB). Gold particles were not labeled on the epithelial lamellae area in the excretory bladder. Bar=1 µg (×25,000)
Fig. 20. The excretory bladder of the worm which reacted with IgG from cats at 3 weeks after infection. Gold particles were rarely labeled on the epithelial lamellae area in the excretory bladder. Bar=1 µg (×25,000)
Fig. 21. The excretory bladder of the worm which reacted with IgG from cats at 4 weeks after infection. Gold particles were slightly labeled on the epithelial lamellae area in the excretory bladder. Bar=1 µg (×25,000)
Fig. 22. The excretory bladder of the worm which reacted with IgG from cats at 12 weeks after infection. Gold particles were predominantly labeled on the epithelial lamellae area in the excretory bladder. Bar=1 µg (×25,000)
Fig. 23. The excretory bladder of the worm which reacted with IgG from cats at 20 weeks after infection. Gold particles were rarely labeled on the epithelial lamellae area in the excretory bladder. Bar=1 µg (×25,000)
Tables
Table 1 Quantitative density of the labeled gold particles in the tegumental syncytium of P. westermani reacted with antibodies (serum)* obtained from cats infected with P. westermani
Table 2 Quantitative density of the labeled gold particles in the secretory granules of P. westermani reacted with antibodies (serum)* obtained from cats infected with P. westermani
Table 3 Quantitative density of the labeled gold particles in the vitelline glands of P. westermani reacted with antibodies (serum)* obtained from cats infected with P. westermani
Table 4 Quantitative density of the labeled gold particles in the epithelial lamella and lumen areas in the caeca of P. westermani reacted with antibodies(serum)* obtained from cats infected with P. westermani
Table 5 Quantitative density of the labeled gold particles in the epithelial lamella areas in excretory bladder of P. westermani reacted with antibodies(serum)* obtained from cats infected with P. westermani
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