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Article contents

Comparative analysis of full-length antigen II/3 from Echinococcus multilocularis and E. granulosus

Published online by Cambridge University Press:  06 April 2009

R. Felleisen
Affiliation:
Institute of Parasitology, University of Berne, Länggass-Strasse 122, P.O. Box 8466, CH-3012 Berne, Switzerland
B. Gottstein
Affiliation:
Institute of Parasitology, University of Berne, Länggass-Strasse 122, P.O. Box 8466, CH-3012 Berne, Switzerland

Summary

The recombinant Echinococcus multilocularis antigen ll/3–10 is one of the most promising tools for immunodiagnosis of alveolar echinococcosis in human patients. Its nucleic acid sequence represents a part of the E. multilocularis gene encoding the metacestode antigen II/3, the former being basically present and expressed in both E. multilocularis and E. granulosus. Most (94%) patients with alveolar echinococcosis respond to infection with a marked anti-II/3–10 IgG synthesis; in contrast, most of the cystic echinococcosis patients do not, for some reason, recognize the recombinant antigen. We tackled this problem by generating cDNA derived from both E. granulosus and E. multilocularis full length II/3 genes, performed by reverse transcription and PCR amplification. Sequence analysis revealed a very high degree of conservation of the primary sequence of the antigen II/3 in both Echinococcus species. cDNA fragments were subcloned and expressed in E. coli as fusion proteins with Schistosoma japonicum glutathione S-transferase. Recombinant proteins were affinity purified and comparatively assessed by ELISA with respect to antibody-binding characteristics. Sera from patients suffering from cystic echinococcosis showed no significant differences in reactivity with the antigens derived from either E. multilocularis or E. granulosus. Therefore, parameters other than some minor differences in the primary sequence seem to be responsible for the lack of antigen II/3 recognition in cystic echinococcosis.

Note

Nucleotide sequence data reported in this paper have been submitted to the GenBank® data base with the accession numbers U05573 and U05574.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1994

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References

Ammann, R. W., Hirsbrunner, R., Cotting, J., Steiger, U., Jacquier, P. & Eckert, J. (1990). Recurrence rate after discontinuation of long-term mebendazole therapy in alveolar echinococcosis (preliminary results). American Journal of Tropical Medicine and Hygiene 43, 506–15.CrossRefGoogle Scholar
Devereux, J. R., Haeberli, P. & Smithies, O. (1984). A comprehensive set of sequence analysis programs for the Vax. Nucleic Acids Research 12, 387–95.CrossRefGoogle ScholarPubMed
Felleisen, H. & Gottstein, B. (1993). Echinococcus multilocularis: molecular and immunochemical characterization of diagnostic antigen II/3–10. Parasitology 107, 335–42.CrossRefGoogle ScholarPubMed
Frohman, M. A. (1990). Rapid amplification of cDNA ends (RACE): user friendly cDNA cloning. Amplifications 5, 1115.Google Scholar
Frosch, P. M., Frosch, M., Pfister, T., Schaad, V. & Bitter-Suermann, D. (1991). Cloning and characterization of an immunodominant major surface antigen of Echinococcus multilocularis. Molecular and Biochemical Parasitology 48, 121–30.CrossRefGoogle Scholar
Frosch, P. M., Geier, C., Kaup, F.-J., Müller, A. & Frosch, M. (1993). Molecular cloning of an echinococcal microtrichal antigen immunoreactive in Echinococcus multilocularis disease. Molecular and Biochemical Parasitology 58, 301–10.CrossRefGoogle ScholarPubMed
Glisin, V., Crkvenjakov, R. & Byus, C. (1974). Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry 13, 2633.CrossRefGoogle ScholarPubMed
Gottstein, B. (1992). Molecular and immunological diagnosis of echinococcosis. Clinical Microbiological Reviews 5, 248–61.CrossRefGoogle ScholarPubMed
Gottstein, B., Deplazes, P. & Aubert, M. (1992). Echinococcus multilocularis: immunological study on the ‘Em2-positive’ laminated layer during in vitro and in vivo post-oncospheral and larval development. Parasitology Research 78, 291–7.CrossRefGoogle ScholarPubMed
Gottstein, B., Jacquier, P., Bresson-Hadni, S. & Eckert, J. (1993). Improved primary immunodiagnosis of alveolar echinococcosis (Echinococcus multilocularis infection of humans) by Em2plus-ELISA. Journal of Clinical Microbiology 31, 373–6.Google Scholar
Hemmings, L. & McManus, D. P. (1991). The diagnostic value and molecular characterisation of an Echinococcus multilocularis antigen gene clone. Molecular and Biochemical Parasitology 44, 5362.CrossRefGoogle ScholarPubMed
Heussler, V. T. & Dobbelaere, D. A. E. (1994). Cloning of a protease gene family of Fasciola hepatica by polymerase chain reaction. Molecular and Biochemical Parasitology 64, 1123.CrossRefGoogle ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Marston, A. O. (1986). The purification of eukaryotic polypeptides synthesized in Escherichica coli. The Biochemical Journal 240, 112.CrossRefGoogle Scholar
Mehlhorn, H., Eckert, J. & Thompson, R. C. A. (1983). Proliferation and metastases formation of larval Echinococcus multilocularis. II. Ultrastructural investigations. Zeitschrift fôr Parasitenkunde 69, 749–63.CrossRefGoogle ScholarPubMed
Môller, N., Gottstein, B., Vogel, M., Flury, K. & Seebeck, T. (1989). Application of a recombinant Echinococcus multilocularis antigen in an enzyme-linked immunosorbent assay for immunodiagnosis of human alveolar echinococcosis. Molecular and Biochemical Parasitology 36, 151–60.CrossRefGoogle Scholar
Olmsted, J. B. (1981). Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. Journal of Biological Chemistry 256, 11955–7.Google ScholarPubMed
Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning. A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.Google Scholar
Smith, D. B. & Johnson, K. S. (1988). Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67, 3140.CrossRefGoogle ScholarPubMed
Towbin, H., Staehelin, T. & Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76, 4350–4.CrossRefGoogle ScholarPubMed
Ullrich, A., Shine, J., Chirgwin, J., Pictet, R., Tischer, E., Rutter, W. J. & Goodman, H. M. (1977). Rat insulin genes: construction of plasrnids containing the coding sequences. Science 196, 1313.CrossRefGoogle ScholarPubMed
Vogel, M., Gottstein, B., Müller, N. & Seebeck, T. (1988). Production of a recombinant antigen of Echinococcus multilocularis with high immunodiagnostic sensitivity and specificity. Molecular and Biochemical Parasitology 31, 117–28.CrossRefGoogle ScholarPubMed
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Comparative analysis of full-length antigen II/3 from Echinococcus multilocularis and E. granulosus
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