• Title/Summary/Keyword: indirect immunoperoxidase stain

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Immunohistochemical study on the antigenicity of each organ structure of Clonorchis sinensis (간흡충 충체의 부위별 항원성에 대한 면역조직화학적 연구)

  • Jin Kim;Jong-Yil Chai;Weon-Gyu Kho;Kyu-Hyuk Cho;Soon-Hyung Lee
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.21-30
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    • 1991
  • 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, rising 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 parenchyme 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.

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Immunohistochemieal study on the antigenicity of body compartments of Payugonimus westermani (폐흡충 충체 부위별 항원성에 대한 면역 조직화학적 연구)

  • Lee, Sun-Hyeong;Seong, Suk-Hwan;Chae, Jong-Il
    • Parasites, Hosts and Diseases
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    • v.27 no.2
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    • pp.109-118
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    • 1989
  • Production of circulating specific antibodies to the lung fluke (Paragenimus 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. Indiret immunoperoxidase(IP) stain technique was applied, using formalin-fked, paraffin- embedded lung tissues of P westermani-infected cats sectioned in 4 Um 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 diaminobensidine(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. The intestinal epithelial border and luminal contents revealed positive staining even at a few concentration of 1 : 4, 000 primary antibody(secondary ab., 1 : 200) whereas the parenchymatous portion showed positive reaction only at higher concentrations than 1'500 (secondary ab., 1 : 200). The results suggest that the specific antibody responses of the host to p. westermani occur most strongly upon the excretes from the intestinal epithelium of the worm and e99s Produced around the worm capsule,

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