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Characterization of excretory-secretory antigens of Fasciola hepatica

Published online by Cambridge University Press:  06 April 2009

D. O. Irving  and
M. J. Howell
D. O. Irving
Affiliation:
Department of Zoology, Faculty of Science, Australian National University, P.O. Box 4, Canberra ACT 2600, Australia
M. J. Howell
Affiliation:
Department of Zoology, Faculty of Science, Australian National University, P.O. Box 4, Canberra ACT 2600, Australia
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Summary

Twenty-one day old Fasciola hepatica were recovered from the livers of infected mice and cultured for 5–7 days in a serum-free medium containing either [C]leucine, [C]isoleucine or [S]methionine, or in a medium containing [C]leucine and serum from a sheep vaccinated with excretory– secretory (ES) antigens of juvenile F. hepatica. All three labelled amino acids were incorporated into fluke proteins. Labelled proteins also appeared in the culture medium. Three major polypeptides detected in the culture media had apparent molecular weights of 26000, 24000 and 23000. All were immunoprecipitated from [C]leucine-labelled culture medium using antisera against fluke somatic antigens raised in rabbits or from sheep vaccinated with ES antigens of juvenile F. hepatica. A polypeptide of molecular weight 27 000 was also prominent in the culture medium when [C]isoleucine was used. This polypeptide was present as a minor component when [C]leucine and [S]methionine were included in the culture media; it did not appear to be immunoprecipitated by the above antisera from [C]leucine-labelled culture medium. In the presence of serum from vaccinated sheep, the ES antigens formed immune complexes which contained the polypeptides mentioned above, together with several higher molecular weight polypeptides. Additionally, a number of minor bands of varying molecular weight were present. After micro-Ouchterlony gel immunodiffusion, 2 precipitin lines formed between the labelled ES antigens and antisera. Electrophoresis of these indicated that the 23000, 24000 and 26000 Dalton labelled polypeptides were present in each. The higher molecular weight and the 27000 Dalton labelled polypeptides were also present in one of the lines.

Type
Research Article
Information
Parasitology , Volume 85 , Issue 1 , August 1982 , pp. 179 - 188
Copyright
Copyright © Cambridge University Press 1982

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References

Chapman, C. B. & Mitchell, G. F. (1982). Fasciola hepaiica: proteolytic cleavage of immunoglobulin. Research in Veterinary Science (in the Press).Google Scholar
Clegg, J. A. & Smith, M. A. (1978). Prospects for the development of dead vaccines against helminths. In Advances in Parasitology, vol. 16 (ed. Lumsden, W. H. R., Muller, R. and Baker, J. R.), pp. 165–218. London, New York and San Francisco: Academic Press.Google Scholar
Cuperlovic, K. (1972). Metabolic antigen of F. hepatica in immunodiagnosis of fascioliasis. Acta Veterinaria (Beograd) 22, 219–22.Google Scholar
Cuperlovic, K. (1975). The incorporation of labelled amino acids into a protein antigen of Fasciola hepatica. Acta Veterinaria (Beograd) 25, 219–27.Google Scholar
Cuperlovic, K. & Lalic, R. (1972). Studies of metabolic antigen activity of adult Fasciola hepatica by complement fixation test. Acta Veterinaria (Beograd) 22, 195–9.Google Scholar
Hajdú, É., Matskási, I. & Juhász, S. (1979). Fasciola hepatica (L., 1758): studies on protease and protease inhibitor activity. Parasitologia Hungarica 12, 21–30.Google Scholar
Howell, M.J. (1979). Vaccination of rats against Fasciola hepatica. Journal of Parasitology 65, 817–19.CrossRefGoogle ScholarPubMed
Irving, D. O. & Howell, M. J. (1982). Preparation and in vitro translation of mRNA from Fasciola hepatica. Molecular and Biochemical Parasitology (in the Press).Google Scholar
Kessler, S. W. (1975). Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody–antigen complexes with protein A. Journal of Immunology 115, 1617–24.CrossRefGoogle ScholarPubMed
Knopf, P. M., Brown, G. V., Howard, R. J. & Mitchell, G. F. (1979). Immunoprecipitation of biosynthetically-labelled products in the identification of antigens of murine red cells infected with the protozoan parasite, Plasmodium berghei. Australian Journal of Experimental Biology and Medical Science 57, 603–15.CrossRefGoogle ScholarPubMed
Kusel, J. R. & MacKenzie, P. E. (1975). The measurement of the relative turnover rates of proteins of the surface membranes and other fractions of Schistosoma mansoni in culture. Parasitology 71, 261–73.CrossRefGoogle ScholarPubMed
Kusel, J. R., MacKenzie, P. E. & McLaren, D. J. (1975). The release of membrane antigens into culture by adult Schistosoma mansoni. Parasitology 71, 247–59.CrossRefGoogle ScholarPubMed
Kusel, J. R., Sher, F. A., Perez, H., Clegg, J. A. & Smithers, S. R. (1975). The use of radioactive isotopes in the study of specific schistosome membrane antigens. In Nuclear Techniques in Helminthology Research, pp. 127–43. Vienna: International Atomic Energy Agency.Google Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Lehner, R. P. & Sewell, M. M. H. (1980). A study of antigens produced by adult Fasciola hepatica maintained in vitro. Parasite Immunology 2, 99–109.CrossRefGoogle Scholar
Locatelli, A. & Beretta, C. (1969). Detection of amino acid N in saline or serum media incubated with Fasciola hepatica and proteolytic activity exerted by liver fluke in vitro. Archivio Veterinario Italiano 20, 385–91.Google ScholarPubMed
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265–75.CrossRefGoogle ScholarPubMed
Movsesijan, M. & Borojević, D. (1973). Antigenic analysis of Fasciola hepatica: extraction and fractionation. In Isotopes and Radiation in Parasitology III, pp. 11–22. Vienna: International Atomic Energy Agency.Google Scholar
Pantelouris, E. M. (1964). Sulfur uptake by Fasciola hepatica L. Life Sciences 3, 1–5.CrossRefGoogle ScholarPubMed
Rajasekariah, G. R., Mitchell, G. F., Chapman, C. B. & Montague, P. E. (1979). Fasciola hepatica: attempts to induce protection against infection in rats and mice by injection of excretory/secretory products of immature worms. Parasitology 79, 393–400.CrossRefGoogle ScholarPubMed
Sandeman, R. M. & Howell, M. J. (1980). In vitro studies of the response of sheep to infection with Fasciola hepatica. Veterinary Parasitology 6, 347–57.CrossRefGoogle Scholar
Sandeman, R. M. & Howell, M. J. (1981). Precipitating antibodies against excretory/secretory antigens of Fasciola hepatica in sheep serum. Veterinary Parasitology 9, 35–46.CrossRefGoogle ScholarPubMed
Sandeman, R. M., Howell, M. J. & Campbell, N. J. (1980). An attempt to vaccinate sheep against Fasciola hepatica using a juvenile fluke antigen sheep antibody complex. Research in Veterinary Science 29, 255–9.CrossRefGoogle ScholarPubMed
Simpkin, K. G., Chapman, C. R. & Coles, G. C. (1980). Fasciola hepatica: a proteolytic digestive enzyme. Experimental Parasitology 49, 281–7.CrossRefGoogle ScholarPubMed
Thorsell, W. & Björkman, N. (1965). Morphological and biochemical studies on absorption and secretion in the alimentary tract of Fasciola hepatica L. Journal of Parasitology 51, 217–23.CrossRefGoogle ScholarPubMed
Wilson, R. J. M. & Ling, I. (1979). Fractionation and characterization of Plasmodium falciparum antigens. Bulletin of the World Health Organization 57, (Suppl. 1), 123–33.Google ScholarPubMed