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Korean J Parasitol > 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.
Abstract

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.

Figures


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.


Tables


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

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