Immunohistochemical study on the antigenicity of each organ structure of <i>Clonorchis sinensis</i>

, Kim, J; , Chai, J Y; , Kho, W G; , Cho, K H; , Lee, S H

doi:1991.29.1.21

Korean J Parasito > Volume 29(1):1991 > Article

Original Article


Korean J Parasitol. 1991 Mar;29(1):21-29. English. Published online Mar 20, 1994. http://dx.doi.org/10.3347/kjp.1991.29.1.21
Copyright © 1991 by The Korean Society for Parasitology


Immunohistochemical study on the antigenicity of each organ structure of Clonorchis sinensis
J Kim,^J Y Chai,W G Kho,^#K H Cho,^ and S H Lee
Department of Parasitology and Institute of Endemic Diseases, University College of Medicine, Seoul 110-460, Korea.


Abstract
An immunohistochemical study was performed to demonstrate comparative antigenicity of each body structure of the liver fluke, Clonorchis sinensis, such as the digestive tract, reproductive organs, excretory system, tegument and suckers. Indirect immunoperoxidase technique was applied, using formalin-fixed and paraffin-embedded sections of C. sinensis as the antigen. Pooled cat sera obtained 10 weeks after an experimental infection with C. sinensis and peroxidase-conjugated goat anti-cat IgG were used as the primary and secondary antibodies, respectively. The intensity of immunohistochemical stain was very sensitive upon the titers of the primary and secondary antibodies, and their optimum dilutions were found to be 1:1,000-1:2,000 and 1:1,000, respectively. The intestinal epithelial cells, intestinal content and excretory bladder showed strong positive coloring reactions even at lower titer (1:2,000) of the primary antibody, whereas the uterine wall and eggs, vitelline glands, and male reproductive organs showed only weak positive reactions despite an increase in the antibody titer (1:1,000). On the other hand, the suckers, tegument, subtegumental cells and other parenchyma portions did not reveal any positive immunoperoxidase reaction at the same antibody titers. From the above results, it is highly suggested that the most potent antigenicity of C. sinensis occur from their excretory-secretory substances originated from the digestive and excretory organs.

Figures


	Fig. 1 Immunoperoxidase staining of sectioned C. Sinensis. Note positive reaction in the in testine(In) (arrow heads), and excretory bladder(EB) (arrow heads). Primary antibody dilution 1:1,000. ×100.


	Fig. 2 Another figure showing a strong positive reaction in the intestine (In) of the worm. Primary antibody dilution 1:500. ×100.


	Fig. 3 Negative control for the immunoperoxidase staining of sectioned C. Sinensis. The section was treated with all steps of immunoperoxidase procedure except using uninfected animal serum instead of infected one (primary antibody). Normal serum dilution 1:1,000. ×100.

Figs. 4-9
Fig. 4. Positive staining is observed in the intestinal epithelium(In) and intestinal content. Primary antibody dilution 1:1,000. ×400.

Fig. 5. Negative control of the same section of C. Sinensis. Normal serum control group, dilution 1:1,000. X400.

Fig. 6. Positive immunoperoxidase reaction is observed on the wall of the excretory bladder(EB). Primary antibody dilution 1:1,000. ×400.

Fig. 7. Negative control at the same sectional level. Normal serum control, dilution 1:1,000. ×400.

Fig. 8. Peroxidase reaction is observed on the uterine wall(UW) and internal substance of the eggs. Primary antibody dilution 1:500. ×400.

Fig. 9. Negative control of the same section. Normal serum control, dilution 1:500. ×400.

Tables


	Table 1 Relative intensity of immunoperoxidase stain at various parts of sectioned C. Sinensis

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