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Translation products of mRNA from infective larvae of Trichinella spiralis include an antigenic polypeptide

Published online by Cambridge University Press:  05 June 2009

Kazuo Sugane ,
Tadashi Matsuura  and
Hideaki Maekawa
Kazuo Sugane
Affiliation:
Department of Parasitology, School of Medicine, Shinshu University, Asahi 3-1-1, Matsumoto City, Japan
Tadashi Matsuura
Affiliation:
Department of Biology, Faculty of Science, Tokyo Metropolitan University, Fukazawa 2-1-1, Setagaya-ku, Tokyo, Japan
Hideaki Maekawa
Affiliation:
National Institute of Health, Kamioosaki 2-10-35, Shinagawa-ku, Tokyo, Japan
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Abstract

Total RNA was extracted from packed infective larvae of Trichinella spiralis by centrifugation through a 5·7 M caesium chloride cushion. Polyadenylated messenger RNA was separated from total RNA in an oligothymidylic acid-cellulose gel column. The in vitro translation of the mRNA, isolated from infective larvae of T. spiralis, was carried out using the rabbit reticulocyte cell-free translation system. Incorporation of 35S-methionine into trichloracetic acid precipitates in the lysate containing mRNA was 5 times greater than that in control. The translation products were analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by autoradiography. Many polypeptides with molecular weights of less than 100 000 were synthesized in the lysate. A T. spiralis positive mouse serum was mixed with translation products to form antigen-antibody complexes, which were then absorbed by Staphylococcus aureus Cowan 1 strain and analysed by autoradiography of SDS-PAGE. An antigenic polypeptide with a molecular weight of 48 000 was demonstrated to react specifically with IgG antibody in T. spiralis positive mouse serum. T. spiralis larvae were cultured in methionine-free medium containing 35S-methionine, and antigenic polypeptides in somatic extracts and ES products were compared with those in translation products by autoradiography of SDS-PAGE. Several polypeptides in ES products and somatic extracts reacted specifically with IgG anitbodies in positive serum. Especially the polypeptide with a molecular weight of 48 000 in ES products strongly reacted with IgG antibody in positive serum.

Keywords

Trichinella spiralismRNA translationantigens
Type
Research Papers
Information
Journal of Helminthology , Volume 61 , Issue 1 , March 1987 , pp. 1 - 8
Copyright
Copyright © Cambridge University Press 1987

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References

REFERENCES

Aviv, H. & Leder, P. (1972) Purification of biologically active messenger RNA by chromatography of oligothymidylic acid-cellulose. Proceedings of the National Academy of Sciences, 69, 1408–1412.Google Scholar
Desavigny, D.H. (1975) In vitro maintenance of Toxocara canis larvae and a simple method for the production of Toxocara ES antigen for use in serodiagnostic tests for visceral larva nugrans. Journal of Parasitology, 61, 781–782.CrossRefGoogle Scholar
Glisin, V., Crkvenjakov, R. & Byus, C. (1974) Ribonucleic acid isolated by cesium chloide centrifugation. Biochemistry, 13, 2633–2637.CrossRefGoogle Scholar
Gould, S.E. (1970) In: Trichinosis in man and animals, editor Gould, S. E., pp. 190–221. Charles C. Thomas Publisher: Springfield, USA.Google Scholar
Hillyer, G.V. & De Rios, I.G. (1979) The enzyme-linked immunosorbent assay (ELISA) for the immunodiagnosis of schistosomiasis. American Journal of Tropical Medicine and Hygiene, 28, 237–241.CrossRefGoogle ScholarPubMed
Irving, D.O. & Howell, M.J. (1981) Preparation and in vitro translation of mRNA from Fasciola hepatica. Molecular and Biochemical Parasitology, 4, 337–348.CrossRefGoogle ScholarPubMed
Irving, D.O. & Howell, M.J. (1982) Characterization of excretory-secretory antigens of Fasciola hepatica. Parasitology, 85, 179–188.CrossRefGoogle ScholarPubMed
Kessler, S.W. (1975) Rapid isolation of antigens from cells with a staphylococcal protein A-antibody absorbent: Parameters of the interaction of antibody-antigen complexes with protein A. Journal of Immunology, 115, 1617–1624.CrossRefGoogle Scholar
Mills, C.K. & Kent, N.H. (1965) Excertions and secretions of Trichinella spiralis and their role in immunity. Experimental Parasitology, 16, 300–310.Google Scholar
O’Beirne, A.J. & Cooper, H.R. (1979) Heterogeneous enzyme immunoassay. Journal of Histochemistry and Cytochemistry, 27, 1148–1162.CrossRefGoogle ScholarPubMed
Parkhouse, R.M.E. & Clark, N.W.T. (1983) Stage specific secreted and somatic antigens of Trichinella spiralis. Molecular and Biochemical Parasitoloty, 9, 319–327.Google ScholarPubMed
Pelham, H.R.B. & Jackson, R.J. (1976) An efficient mRNA dependent translation system from reticulocyte lysate. European Journal of Biochemistry, 67, 247–256.Google Scholar
Silberstein, D.S. & Despommier, D.D. (1984) Antigens from Trichinella spiralis that induce a protective response in the mouse. Journal of Immunology, 132, 898–904.Google Scholar
Sugane, K. & Oshima, T. (1982) Eosinophilia, granuloma formation and migratory behaviour of larvae in the congenitally athymic mouse infected with Toxocara canis. Parasite Immunology, 4, 307–318.Google Scholar
Suzuki, Y. & Kobayashi, A. (1985) Detection of circulating antigens by latex particles coated with anti-Toxoplasma antibodies during acute infactions with Toxoplasma gondii in mice. Japanese Journal of Parasitology, 34, 149–154.Google Scholar
Weber, K. & Osborn, M. (1969) The reliability of molecular weight determainations by dodecyl sulfate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry, 244, 4406–4412.CrossRefGoogle ScholarPubMed