| Home | E-Submission | Sitemap | Contact us |  
Korean J Parasito Search


Parasit Host Dis > Volume 27(2):1989 > Article

Original Article
Korean J Parasitol. 1989 Jun;27(2):109-117. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1989.27.2.109
Copyright © 1989 by The Korean Society for Parasitology
Immunohistochemical study on the antigenicity of body compartments of Paragonimus westermani
S H Lee,S H Sung,* and J Y Chai
Department of Parasitology and Institute of Endemic Diseases, Department of Thoracic Surgery, College of Medicine, Seoul National University, Seoul 110-460, Korea.

Production of circulating specific antibodies to the lung fluke (Paragonimus westermani) by its host is well known and used in various kinds of immunodiagnostic methods. However, it has not been well documented which compartments (or structures) of the lung fluke are most responsible for the production of specific antibodies. The present immunohistochemical study was undertaken to demonstrate the antigenicity of each body compartment of P. westermani such as suckers, tegument, spines, vitelline glands, intestine, reproductive organs (male and female), and eggs. Indirect immunoperoxidase (IP) stain technique was applied, using formalin-fixed, paraffin-embedded lung tissues of P. westermani-infected cats sectioned in 4 µm thickness as the antigen and cat antisera (11-20 weeks of infection) as the primary antibody. Peroxidase-conjugated goat anti-cat IgG was used as the secondary antibody and diaminobenzidine (DAB) as the coloring agent. Strong yellow or yellowish brown staining was regarded positive. The primary and secondary antibody dilutions were made at 1:500-1:2,000 and 1:200-1:500 respectively, and IP stain was repeated 10 times for each dilution. A consistent result obtained was that the intestinal epithelial border, intestinal content, vitelline glands, and eggs scattered around the worm capsule showed strong positive staining, while uterine eggs and some parenchymal portions showed weak positive reaction. On the other hand, the suckers, tegument, spines, subtegumental cells, cytoplasm of intestinal epithelial cells, male reproductive organs, and ovary revealed negative staining. The body compartments showing higher antigenicity were, in the decreasing order, the intestinal epithelial border, intestinal content, eggs in the worm capsule, vitelline glands, uterine eggs, and parenchymatous portions.


Figs. 1-6
Fig. 1. Negative control immunoperoxidase (IP) staining of P. westermani (×100). The peroxidase-conjugated goat anti-cat IgG (secondary antibody) step was omitted for this slide. The eggs (arrows) outside of the worm consist of yellowish egg shell and blue-stained yolk or germ cell.

Fig. 2. Strong positive(⧺) IP stain of intestinal luminal border of P. westermani (×100). Infected cat serum (11 wk infection) diluted to 1: 1,000 and peroxidase-conjugated goat anti-cat IgG to 1:200, 30 minutes incubation each. Vitelline follicles (arrows) also show positive stain.

Fig. 3. Another strong positive(⧺) stain (×100). Infected cat serum (11 wk) diluted to 1:1,000 and peroxidase-conjugated goat anti-cat IgG to 1:200, 30 minutes incubation each. Note positive stain of intestinal content, lining epithelial border, and vitelline glands(arrows). Ovary (OV) shows negative stain.

Fig. 4. An oblique section of P. westermani showing positive(⧺) IP staining of the intestine (×100). The same group as in Fig. 3. Positive and negative stains, however, are not so sharply demarcated at the epithelial lining portion of the intestine.

Fig. 5. Positive IP staining of P. westermani eggs (arrows) in a worm capsule of a cat lung (×100). Infected cat serum (12 wk) diluted to 1:1,000 and peroxidase-conjugated goat anti-cat IgG to 1:200, 30 minutes incubation each.

Fig. 6. Positive IP staining of P. westermani eggs (arrows) in another section (×100). Infected cat serum (11 wk) diluted to 1:500 and peroxidase-conjugated goat anti-cat IgG to 1:200, 30 minutes incubation each.


Table 1
Checkerboard titration of primary and secondary antibodies for indirect immunoperoxidase staining of P. westermani

Table 2
Relative intensity of peroxidase staining at various compartments of sectioned P. westermani

2. Cho KM, et al. Yonsei Rep Trop Med 1976;7:26–39.
3. Cho SY, Hong ST, Rho YH, Choi SY, Han YC. Application of micro-ELISA in serodiagnosis of Human paragonimiasis. Korean J Parasitol 1981;19(2):151–156.
4. Cho SY, Kim SI, Kang SY, Kong Y, Han SK, Shim YS, Han YC. Antibody changes in paragonimiasis patients after praziquantel treatment as observed by ELISA and immunoblot. Korean J Parasitol 1989;27(1):15–21.
5. Cho SY, Lee DK, Kang SY, Kim SI. [An Epidemiological Study Of Human Paragonimiasis By Means Of Micro-ELISA]. Korean J Parasitol 1983;21(2):246–256.
6. Choi WY, Lee OR. [Agar-Gel Precipitin Reactions In Experimental Paragonimiasis]. Korean J Parasitol 1981;19(2):101–108.
7. Choi WY, Yoo JE, Nam HW, Choi HR. [Purification of antigenic proteins of Paragonimus westermani and their applicability to experimental cat paragonimiasis]. Korean J Parasitol 1986;24(2):177–186.
9. Damonneville M, Pierce RJ, Verwaerde C, Capron A. Allergens of Schistosoma mansoni. II. Fractionation and characterization of S. mansoni egg allergens. Int Arch Allergy Appl Immunol 1984;73(3):248–255.
11. Hillyer GV, Serrano AE. The antigens of Paragonimus westermani, Schistosoma mansoni, and Fasciola hepatica adult worms. Evidence for the presence of cross-reactive antigens and for cross-protection to Schistosoma mansoni infection using antigens of Paragonimus westermani. Am J Trop Med Hyg 1983;32(2):350–358.
12. Hunter GW 3rd, Ritchie LS, Pan C, Lin S, Sugiura S, Nagano K, Yokogawa M. Immunological studies, II. Intradermal tests and their application in the field for the detection of schistosomiasis japonica, paragonimiasis and clonorchiasis. Mil Med 1958;122(2):85–96.
13. Imai J. Trop Med 1979;21(2):45–55.
14. Joo KH, Ahn H, Chung MS, Rim HJ. Demonstration of species-specific and cross reactive components of Paragonimus westermani crude worm antigen by EITB. Korean J Parasitol 1989;27(1):9–14.
15. Joo KH, Hong SC, Chung MS, Rim HJ. [Analysis of antigenic specificities of Paragonimus westermani developmental stages using immunoblot technique]. Korean J Parasitol 1989;27(1):1–7.
17. Kim SH, Kong Y, Kim SI, Kang SY, Cho SY. Immunoblot observation of antigenic protein fractions in Paragonimus westermani reacting with human patients sera. Korean J Parasitol 1988;26(4):239–243.
18. Kim SI, Kang SY, Cho SY. [On The Applicability Of Partially Purified Antigenic Preparations Of Paragonimus Westermani]. Korean J Parasitol 1983;21(2):257–264.
19. Kim SI, Ko EK, Kang SY, Cho SY. [Antigenicity of the soluble egg antigen of Paragonimus westermani]. Korean J Parasitol 1986;24(1):49–54.
20. Kobayashi F, et al. Jpn J Parasitol 1985;34(4):253–260.
21. Lee OR, Chang JK. [ELISA of paragonimiasis in cat by crude and purified antigens of Paragonimus westermani]. Korean J Parasitol 1986;24(2):187–193.
22. Lee OR, Choi WY. [Comparison Of Agar-Gel Diffusion Tests, Counterimmunoelectrophoresis And Enzyme-Linked Immunosorbent Assay In The Sera Of Skin Test Positives For Paragonimiasis]. Korean J Parasitol 1983;21(2):270–280.
23. von Lichtenberg F, Bawden MP, Shealey SH. Origin of circulating antigen from the schistosome gut. An immunofluorescent study. Am J Trop Med Hyg 1974;23(6):1088–1091.
24. Matsuda H, et al. Jpn J Parasitol 1984;33(3):163–170.
25. Nash TE. Localization of the circulating antigen within the gut of Schistosoma mansoni. Am J Trop Med Hyg 1974;23(6):1085–1087.
26. Ohara H, et al. Jpn J Parasitol 1985;34(4):245–252.
27. Pelley RP, Pelley RJ, Hamburger J, Peters PA, Warren KS. Schistosoma mansoni soluble egg antigens. I. Identification and purification of three major antigens, and the employment of radioimmunoassay for their further characterization. J Immunol 1976;117(5 Pt 1):1553–1560.
28. Sadun EH, Buck AA, Walton BC. The diagnosis of paragonimiasis westermani using purified antigens in intradermal and complement fixation tests. Mil Med 1959;124(3):187–195.
29. Sawada T, Takei K, Voneyama K. Studies On The Immunodiagnosis Of Paragonimiasis. I. The Precipitin Reaction With Crude And Fractionated Antigens. J Infect Dis 1964;114:311–314.
30. Soh CT, et al. Yonsei Rep Trop Med 1985;16(1):1–10.
31. Sugiyama H, Sugimoto M, Akasaka K, Horiuchi T, Tomimura T, Kozaki S. Characterization and localization of Paragonimus westermani antigen stimulating antibody formation in both the infected cat and rat. J Parasitol 1987;73(2):363–367.
32. Sun T, Gibson JB. Antigens of Clonorchis sinensis in experimental and human infections. Am J Trop Med Hyg 1969;18(2):241–252.
33. Yogore MG, et al. Am J Trop Med Hyg 1965;14(4):586–591.
35. Yokogawa M, et al. Jpn J Parasitol 1962;11(2):117–122.
Editorial Office
Department of Molecular Parasitology, Samsung Medical Center, School of Medicine, Sungkyunkwan University,
2066 Seobu-ro, Jangan-gu, Suwon 16419, Gyeonggi-do, Korea.
Tel: +82-31-299-6251   FAX: +82-1-299-6269   E-mail: kjp.editor@gmail.com
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Copyright © 2023 by The Korean Society for Parasitology and Tropical Medicine.     Developed in M2PI