The sensitivity and specificity of crude and affinity-purified antigens of Clonorchis sinensis obtained from the infected rabbits were studied. Stage-specific antigenic proteins from the eggs, metacercariae and adult worms were characterized by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunosorbent assay (ELISA) The results were as follows: The antibody-binding antigen (ABA) purified from whole worm crude antigen (WWA) by CNBr-activated Sepharose 4B affinity chromatography made 4 specific bands against rabbit anti-sera on Ouchterlony gel diffusion plate, while WWA made 7 bands. Major WWA protein bands by SDS-PAGE were found at 16,300-18,500 and 28,000-29,000 daltons, while major ABA protein bands were at 18,000-21,000 and 29,000-31,000 daltons. The reactivity of ABA with rabbit anti-sera in ELISA was remarkably less sensitive than that of WWA.
Molecular weights of egg antigen (EGA), metacercarial antigen (MEA) and adult worm antigen (WWA) of C. sinensis ranged from 15,000-200,000 daltons, 15,000-100,000 daltons and 11,000-80,000 daltons, respectively. Major WWA proteins consisted mainly of polypeptide bands of low molecular weight, less than 31,000 daltons, while those of EGA and MEA consisted of higher molecular weights than 30,000 daltons.
The ELISA reactivities of WWA to rabbit anti-sera were remarkably greater than those of MEA. EGA showed negative reaction throughout the experiments. WWA showed higher optical density (O.D.) than 1.0, when reacted with rabbit anti-sera obtained at 4-6 weeks after the infection. In the rabbit anti-sera later than 12 weeks after the infection, the O.D. reacting with WWA showed a plateau without variation. MEA showed relatively low O.D. values (<0.6), when reacted with anti-sera from lightly infected groups throughout the experiments, although there were some weak positive cases (O.D.>0.6) in heavily infected groups. MEA reacted with rabbit anti-sera showed negative results on Ouchterlony gel diffusion plates. Summarizing the above results, it is suggested that the whole worm antigen prepared from the adult worms of C. sinensis is most highly antigenic. However, this antigen might reveal cross reactions with other trematodes such as Paragonimus westermani, therefore, purification of antigenic proteins from the crude antigen is essential to increase the sensitivity and specificity for the immunodiagnosis of clonorchiasis.
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Molecular cloning and immunological characterization of phosphoglycerate kinase from Clonorchis sinensis Sung-Jong Hong, Kee-Young Seong, Woon-Mok Sohn, Kye-Yong Song Molecular and Biochemical Parasitology.2000; 108(2): 207. CrossRef
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Immune reactions between excretory-secretory antigens and specific antibodies of Clonorchis sinensis before and after praziquantel treatment in experimentally infected rabbits S I Kim The Korean Journal of Parasitology.1994; 32(1): 35. CrossRef
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Immunohistochemical study on the antigenicity of each organ structure of Clonorchis sinensis J Kim, J Y Chai, W G Kho, K H Cho, S H Lee The Korean Journal of Parasitology.1991; 29(1): 21. CrossRef
Analysis of Clonorchis sinensis antigens and diagnosis of clonorchiasis using monoclonal antibodies T S Yong, K Im, P R Chung The Korean Journal of Parasitology.1991; 29(3): 293. CrossRef
ELISA of rat sera infected with Paragonimus iloktsuenensis B K Lim, O R Lee, H S Nam The Korean Journal of Parasitology.1990; 28(4): 207. CrossRef
By affinity chromatography using a monoclonal antibody as ligand, Kim et al. (1986) purified a protein fraction in cystic fluid of Taenia solium metacestodes (CF). In this study, the biochemical properties of the purified protein were characterized. Discontinuous-polyacrylamide gel electrophoresis (disc-PAGE) of the protein at 4.5~10% separating gel concentration showed its molecular weight (MW) to be 150 kilodalton(kDa) in non-denatured state, while denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that it was composed of 3 different subunits with respective MW of 15, 10 and 7 kDa.
Subunit of 7 kDa was shown to be linked to other subunits by disulfide bonds. Isoelectric point of the protein was pH 6.8. The protein was relatively heat-stable for immunologic analysis. These properties indicated that the protein, comprising about 70% of total content in CF, had similar biochemical characters with antigen B of Oriol et al.(1971) in hydatid cyst fluid (HF)
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Component proteins in cystic fluid of Taenia solium metacestodes collected surgically from neurocysticercosis patients Y Kong, S Y Kang, S Y Cho The Korean Journal of Parasitology.1990; 28(2): 101. CrossRef
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Specific or non-specific cytolytic processes of free-living amoebae causing meningoencephalitis have been emphasized and the cytolytic ability related to hydrolases in Entamoeba sp.
and Naegleria sp. has also been reported since the latter half of 1970's. However, no information on hydrolase activities in Acanthamoeba sp. is available. Hydrolases in Acanthamoeba culbertsoni, a pathogenic species of free-living amoebae, were assayed and compared with those in a non-pathogenic species, A. royreba. Pathogenicity of these two species was confirmed through experimental infection to BALB/c mice. Hydrolase activities and cytotoxic effects between pathogenic and non-pathogenic species were compared in the trophozoites cultured in CGV media and in CHO cell line, respectively. The results are summarized as follows: The mice infected with A. culbertsoni were all dead 15 days after nasal inoculation, and the mean survival time was 8.5 days. Also the mice infected with this pathogenic species mani fested typical meningoencephalitis, whereas the mice infected with A. royreba did not. Hydrolases detected both in the cell extracts and culture media were acid phosphatase, beta- N-acetyl galactosaminidase, beta-N-acetyl glucosaminidase, alpha-mannosidase, neutral proteinase and acid proteinase, all of which were detected with remarkably higher rate in A.culbertsoni than in A. royreba. A.
culbertsoni revealed strong cytotoxicity for the target CHO cells, whereas A. royreba did not show any specific cytotoxicity. About 80 % of the target cells mixed with A.
culbertsoni were dead 48 hours after cultivation, and more than 95% of the target cells were dead 72 hours after cultivation. Hydrolase activities in A. culbertsoni cultured with the target cell line were assayed according to the culture time. The activities of acid phosphatase, beta-N-acetyl glucosaminidase, beta-N-acetyl glucosaminidase, alpha-mannosidase and acid proteinase in this pathogenic amoeba were detected higher in amoeba extracts than in culture media up to 120 hours after cultivation, but after 120 hours of cultivation those activities were detected higher in culture media than in the amoeba lysates. Neutral proteinase activity in A.
culbertsoni increased more in EBSS medium than in the lysate specimens although the activity in the extracts was generally steady according to the cultivation time.
Summarizing the above results, it is concluded that there were differences in hydrolase activities between pathogenic A. culbertsoni and non-pathogenic A. royreba, and that some hydrolase activities were detected remarkably higher in A.
culbertsoni which revealed strong cytotoxicity to the target CHO cell line.
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As a series of systematic classification of paramphistomes, the worms in the rumen and reticulum of 310 Korean cattle slaughtered at Chonju abattoir were collected from February 1986 to June 1987 and were classified by morphology of the worms. Afterwards, the karyotype of Fischoederius cobboldi (Poirier, 1883), which is a very rare species in Korean cattle, was studied with germ cells of the worm by means of modified air-drying method. The chromosome numbers in the haploid and diploid cells of 315 F. cobboldi were n=9 and 2n=18, respectively. The meiotic divisions were observed frequently; 1,904 haploid and 49 diploid cells were recognized. Nine pairs of mitotic chromosomes were homologous in the metaphase stage and the chromosomes were composed of seven medium-sized metacentrics (m) or submetacentrics (sm) and two small-sized submetacentrics (sm). While, meiotic metaphases were composed of seven medium and two small-sized chromosomes. The 3rd, 4th, 2nd and 5th pairs of chromosomes was metacentric having centromere indices of 40.4 per cent, 40.0 per cent, 39.7 per cent and 38.9 per cent, respectively, and the remaining ones were submetacentric with centromere indices from 32.4 per cent to 36.2 per cent. As a series of C-banding method, C-band was shown in centromeric region from all of the haploid germ cells, except chromosome No. l which included heterochromatin at the tip region. Chromosomes No. 4, 6 and 8 showed remarkable C-band distinguished from others.
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On October 26, 1987, the authors encountered a case of human thelaziasis involving both eyes. The case in Chollapuk-do, Korea, a farmer aged 23, complained of mild lacrimation for 3 weeks followed by 1 week duration of foreign body sensation in both eyes prior to examination of eyes. The conjunctivae of both eyes appeared slightly reddened. No other abnormalities were noticed. Total 6 worms, one male and two females from each eye, were removed, parasitologically studied, and identified as Thelazia callipaeda Railliet and Henry, 1910.
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