Hostname: page-component-78c5997874-lj6df Total loading time: 0 Render date: 2024-11-03T00:35:51.766Z Has data issue: false hasContentIssue false

Analysis of Opisthorchis viverrini antigens by immunoprecipitation and polyacrylamide gel electrophoresis

Published online by Cambridge University Press:  06 April 2009

S. Wongratanacheewin
Affiliation:
Department of Microbiology, Faculty of Science, and Hospital for Tropical Diseases
R. Chawengkirttikul
Affiliation:
Department of Microbiology, Faculty of Science, and Hospital for Tropical Diseases
D. Bunnag
Affiliation:
Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand
S. Sirisinha
Affiliation:
Department of Microbiology, Faculty of Science, and Hospital for Tropical Diseases

Summary

Antigenicity of adult Opisthorchis viverrini, metacercariae and eggs was investigated by radio-immunoprecipitation and polyacrylamide gel electrophoresis using sera from patients with opisthorchiasis and from hamsters experimentally infected with 0. viverrini metacercariae. 125I-labelled aqueous somatic extract and in vitro metabolic products of adult worms, and the extracts of metacercariae and eggs were immunoprecipitated, electrophoresed and autoradiographed and major antigens were characterized. Although components with Mr of > 116, 89, 78 and 20 kD appeared to be specifically associated with the somatic extract of adult fluke, the 89 kD protein was the most prominent component found in the in vitro culture fluid of adult worms and the metacercarial extract. Hamsters, experimentally infected with 0. viverrini, also showed the presence of antibody reactive with the 89 kD component within 15 days of infection. The potential of this antigen in immunodiagnosis is discussed.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1988

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Boonpucknavig, S., Kurathong, S. & Thamavit, W. (1986). Detection of antibodies in sera from patients with opisthorchiasis. Journal of Clinical and Laboratory Immunology 19, 135–7.Google ScholarPubMed
Feldheim, W. & Knobloch, J. (1982). Serodiagnosis of opisthorchis viverrini infestation by an enzyme immunoassay. Tropenmedizin und Parasitologie 33, 810Google Scholar
Flavell, D. J. (1981). The in vitro effects of serum on the adults, metacercariae and eggs of Opisthorchis viverrini. Transactions of the Royal Society for Tropical Medicine and Hygiene 75, 825–31.CrossRefGoogle ScholarPubMed
Flavell, D. J. & Flavell, S. U. (1986). Opisthorchis viverrini: pathogenesis of infection in immunodeprived hamsters. Parasite Immunology 8, 455–66.CrossRefGoogle ScholarPubMed
Greenwood, F. C., Hunter, W. M. & Glover, J. S. (1963). The preparation of 131I-labelled human growth hormones of high specific radioactivity. The Biochemical Journal 89, 114–23.CrossRefGoogle Scholar
Ivarie, R. D. & Jones, P. P. (1979). A rapid sensitive assay for specific protein synthesis in cells and in cell-free translations: use of Staphylococcus aureus as an adsorbent for immune complexes. Analytical Biochemistry 97, 2435.CrossRefGoogle ScholarPubMed
Janechaiwat, J., Tharavanij, S., Vajrasthira, S. & Chaicumpa, W. (1980). The immunological diagnosis of human opisthorchiasis and humoral immune response to opisthorchis infection in the hamster. Journal of the Medical Association of Thailand 63, 439–47.Google ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. S. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265–75.CrossRefGoogle ScholarPubMed
Puengtomwatanakul, S. & Sirisinha, S. (1986). Impaired biliary secretion of immunoglobulin A in vitamin A-deficient rats. Proceedings of the Society for Experimental Biology and Medicine 182, 437–42.CrossRefGoogle ScholarPubMed
Ruppel, A., Boonpucknavig, S. & Hempelmann, E. (1985). Unusual protein pattern of Opisthorchiasis viverrini. Journal of Helminthology 59, 347–9.CrossRefGoogle Scholar
Seqrest, J. P. & Jackson, R. L. (1972). Molecular weight determination of glycoproteins by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Methods in Enzymology 28, 5463.CrossRefGoogle Scholar
Sirisinha, S. (1986). Immunodiagnosis of human liver fluke infections. Asian Pacific Journal of Allergy and Immunology 4, 81–8.Google ScholarPubMed
Sirisinha, S., Tuti, S., Tawatsin, A. & Rattanasiriwilai, W. (1982). Lack of evidence for protective immunity in hamsters infected with Opisthorchis viverrini. In Proceedings of the 25th Seameo-Tropmed Seminar, 19–25 October, 1982 (ed. Mak, J. W. and Yong, H. S.), pp. 91100. Malaysia.Google Scholar
Sirisinha, S., Tuti, S., Vichasri, S. & Tawatsin, A. (1983). Humoral immune responses in hamsters infected with Opisthorchis viverrini. Southeast Asian Journal of Tropical Medicine and Public Health 14, 243–51.Google ScholarPubMed
Sirisinha, S., Puengtomwatanakul, S., Sobhon, P., Saitongdee, P., Wongpayabal, P., Mitranonde, V., Radomyos, P., Bunnag, D. & Harinasuta, T. (1984). Alterations of the surface tegument of Opisthorchis viverrini exposed to praziquantel in vitro and in vivo. Southeast Asian Journal of Tropical Medicine and Public Health 15, 95103.Google ScholarPubMed
Srivatanakul, P., Viyanant, V., Kurathong, S. & Tiwawech, D. (1985). Enzyme-linked immunosorbent assay for detection of Opisthorchis viverrini infection. Southeast Asian Journal of Tropical Medicine and Public Health 16, 234–9.Google ScholarPubMed
Tuti, S., Vichasri, S. & Sirisinha, S. (1982). Effect of culture media on production of excretory-secretory and egg output of Opisthorchis viverrini in vitro. Journal of Parasitology 68, 892–7.CrossRefGoogle ScholarPubMed
Weber, K. & Osborn, M. (1969). The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244, 4406–11.CrossRefGoogle ScholarPubMed