Hostname: page-component-cd9895bd7-hc48f Total loading time: 0 Render date: 2024-12-28T13:19:32.974Z Has data issue: false hasContentIssue false
  • English
  • Français

Echinococcus multilocularis: molecular and immunochemical characterization of diagnostic antigen II/3-10

Published online by Cambridge University Press:  06 April 2009

R. Felleisen  and
B. Gottstein
R. Felleisen
Affiliation:
Institute of Parasitology, University of Berne, Länggasse-Strasse 122, 3001 Berne, Switzerland
B. Gottstein
Affiliation:
Institute of Parasitology, University of Berne, Länggasse-Strasse 122, 3001 Berne, Switzerland
Get access Rights & Permissions [Opens in a new window]

Summary

A recombinant Echinococcus multilocularis antigen (II/3–10), which had previously been shown to exhibit immunodiagnostic characteristics highly specific for human alveolar echinococcosis, and the corresponding native parasite antigen, were further characterized with immunochemical and molecular biological methods. Immunoblot analysis using a polyclonal antiserum raised in rabbits against the recombinant protein, and subsequent Northern hybridization analysis, revealed that the native antigen was expressed by E. multilocularis at the adult as well as at the metacestode stage. In metacestodes, the antigen was shown by using indirect immunofluorescence and the same antiserum to be localized within the germinal layer and membrane structures of developing protoscolices. Electrophoretic analyses revealed remarkable differences in the apparent molecular weight of the antigen under reducing and non-reducing conditions. In further immunoblot analyses, anti-II/3–10 antibodies identified the corresponding epitopes on bands with identical Mr in all E. multilocularis isolates investigated (European, Asian and North American). By Southern hybridization analyses of the respective gene, phylogenetically related sequences were shown to be present in other helminth species such as E. granulosus and several Taenia spp. In the same respect, immunoblotting revealed that anti-II/3–10 antibodies reacted with antigens of different Mr from various E. granulosus isolates and some other cestode species, indicating the presence of shared and thus potentially cross-reacting epitopes. The relevance of these findings for the immunodiagnostic performance of the recombinant antigen is discussed.

Keywords

Echinococcus multilocularisTaeniid cestodesimmunodiagnosisrecombinant antigen
Type
Research Article
Information
Parasitology , Volume 107 , Issue 3 , September 1993 , pp. 335 - 342
Copyright
Copyright © Cambridge University Press 1993

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

Feinberg, A. P. & Vogelstein, B. (1983). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 132, 6–13.CrossRefGoogle ScholarPubMed
Feinberg, A. P. & Vogelstein, B. (1984). Addendum: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 137, 266.Google ScholarPubMed
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 ScholarPubMed
Glisin, V., Crkvenjakov, R. & Byus, C. (1974). Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry 13, 2633.CrossRefGoogle ScholarPubMed
Gottstein, B. (1991). Echinococcus multilocularis: antigenic variance between different parasite isolates. Parasitology Research 77, 359–61.CrossRefGoogle ScholarPubMed
Gottstein, B. & Nash, T. E. (1991). Antigenic variation in Giardia lamblia: infection of congenitally athymic nude and scid mice. Parasite Immunology 13, 649–59.CrossRefGoogle ScholarPubMed
Gottstein, B., Deplazes, P. & Aubert, M. (1992). Echinococcus multilocularis: immunological study on the ‘Em2-positive’ laminated layer during in vitro and in viva post-oncospheral and larval development. Parasitology Research 78, 291–7.CrossRefGoogle Scholar
Gottstein, B., Jacquier, P., Bresson-Hadni, S. & Eckert, J. (1993). Improved primary immunodiagnosis of alveolar echinococcosis in humans by an enzyme- linked immunosorbent assay using the Em2plus-antigen. Journal of Clinical Microbiology 31, 373–6.CrossRefGoogle ScholarPubMed
Gottstein, B. & Mowatt, M. R. (1991). Sequencing and characterization of an Echinococcus multilocularis DNA probe and its use in the polymerase chain reaction. Molecular and Biochemical Parasitology 44, 183–94.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
Lehrach, H., Diamond, D., Wozney, J. M. & Boetker, H. (1977). RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry 16, 4743–51.CrossRefGoogle ScholarPubMed
Müller, 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
Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: a Laboratory Manual, 2nd edn.Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
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 plasmids 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 Biological Parasitology 31, 117–25.CrossRefGoogle ScholarPubMed