Hostname: page-component-cd9895bd7-dk4vv Total loading time: 0 Render date: 2024-12-27T10:02:29.975Z Has data issue: false hasContentIssue false
  • English
  • Français

Fractionation and antigenic characterization of organelles of Eimeria tenella sporozoites

Published online by Cambridge University Press:  06 April 2009

U. Kawazoe ,
F. M. Tomley  and
J. A. Frazier
U. Kawazoe
Affiliation:
Institute for Animal Health, Houghton Laboratory, Houghton, Huntingdon, Cambs PE17 2DA
F. M. Tomley
Affiliation:
Institute for Animal Health, Houghton Laboratory, Houghton, Huntingdon, Cambs PE17 2DA
J. A. Frazier
Affiliation:
Institute for Animal Health, Houghton Laboratory, Houghton, Huntingdon, Cambs PE17 2DA
Get access Rights & Permissions [Opens in a new window]

Summary

Sporozoites of Eimeria tenella were disrupted by sonication and subcellular fractions were separated by sucrose gradient ultracentrifugation. Fractions from gradients were characterized by electron microscopical appearance and their polypeptide and antigenic profiles determined by PAGE and Western blotting with antisera to sporozoites and 1st- and 2nd-generation merozoites. Fractions containing micronemes, rhoptries or membranes showed markedly different polypeptide content and antigenic reactivity. Microneme epitopes were strongly conserved between sporozoites and 2nd-generation merozoites whereas the majority of rhoptry epitopes and many membrane epitopes were sporozoite specific. The only polypeptide of sporozoites which was strongly recognized by antisera raised to 1st generation merozoites was a microneme antigen of molecular weight approximately 100 kDa.

Keywords

Eimeria tenellasporozoitesorganellesantigens
Type
Research Article
Information
Parasitology , Volume 104 , Issue 1 , February 1992 , pp. 1 - 9
Copyright
Copyright © Cambridge University Press 1992

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

Adams, J. U., Hudson, D. E., Torii, M., Ward, G. E., Wellems, T. E., Aikama, M. & Miller, L. H. (1990). The Duffy receptor family of Plasmodium knowlesi is located within the micronemes of invasive malaria merozoites. Cell 63, 141–53.CrossRefGoogle ScholarPubMed
Augustine, P. C., Danforth, H. D. & Mcandrew, S. J. (1988). Monoclonal antibodies reveal antigenic differences in refractile bodies of avian Eimeria sporozoites. Journal of Parasitology 74, 653–9.CrossRefGoogle ScholarPubMed
Bushell, G. R., Ingram, L. T., Fardoulys, C. A. & Cooper, J. A. (1988). An antigenic complex in the rhoptries of Plasmodium falciparum. Molecular and Biochemical Parasitology 28, 105–12.CrossRefGoogle ScholarPubMed
Chapman, H. D. (1978). Studies on the excystation of different species of Eimeria in vitro. Zeitschrift für Parasitenkunde 56, 115–21.CrossRefGoogle ScholarPubMed
Chobotar, B. & Scholtysek, F. (1982). Ultrastructure. In The Biology of the Coccidia, pp. 101–165. (ed. Long, P. L.) London: Edward Arnold.Google Scholar
Danforth, H. D. & Augustine, P. C. (1989). Eimeria tenella: Use of a monoclonal antibody in determining the intracellular fate of the refractile body organelles and the effect on in vitro development. Experimental Parasitology 68, 1–7.CrossRefGoogle ScholarPubMed
Danforth, H. D. & McAndrew, S. J. (1987). Hybridoma antibody characterization of stage-specific and stage-cross-reactive antigens of Eimeria tenella. Journal of Parasitology 73, 985–92.CrossRefGoogle ScholarPubMed
Dubremetz, J-F. & Dissous, C. (1980). Characteristic proteins of micronemes and dense granules from Sarcocystis tenella zoites (Protozoa, coccidia). Molecular and Biochemical Parasitology 1, 279–89.Google Scholar
Dubremetz, J-F., Ferreira, E. & Dissous, C. (1989). Isolation and partial characterisation of rhoptries and micronemes from Eimeria nieschulzi zoites (Sporozoa, Coccidia). Parasitology Research 75, 449–54.CrossRefGoogle ScholarPubMed
Dzandu, J. K., Deh, M. E., Barratt, D. L. & Wise, G. E. (1984). Detection of erythrocyte membrane proteins, sialoglycoproteins, and lipids in the same polyacrylamide gel using a double-staining technique. Proceedings of the National Academy of Sciences, USA 81, 1733–7.CrossRefGoogle ScholarPubMed
Entzeroth, R., Dubremetz, J-F., Hodick, D. & Ferreira, E. (1986). Immunoelectron microscopic demonstration of exocytosis of dense granule contents into the secondary parasitophorous vacuole of Sarcocystis muris (Protozoa, Apicomplexa). European Journal of Cell Biology 41, 182–8.Google ScholarPubMed
Iams, K. P., Young, J. R., Nene, V., Desai, J., Webster, P., Ole-Moi Yoi, O.K. & Musoke, A. J. (1990). Characterisation of the gene encoding a 104-kilodalton microneme-rhoptry protein of Theileria parva. Molecular and Biochemical Parasitology 39, 47–60.CrossRefGoogle ScholarPubMed
Leriche, M. A. & Dubremetz, J. F. (1991). Characterization of the protein contents of rhoptries and dense granules of Toxoplasma gondii tachyzoites by subcellular fractionation and monoclonal antibodies. Molecular and Biochemical Parasitology 45, 249–60.CrossRefGoogle ScholarPubMed
Long, P. L., Joyner, L. N., Millard, B. J. & Norton, C. C. (1976). A guide to laboratory techniques used in the study and diagnosis of avian coccidiosis. Folia Veterinaria Latina 6, 201–17.Google Scholar
McDonald, V., Rose, M. E. & Jeffers, T. K. (1986). Eimeria tenella: Immunogenicity of the first generation of schizogony. Parasitology 93, 1–7.CrossRefGoogle ScholarPubMed
McDonald, V., Wisher, M. H., Rose, M. E. & Jeffers, T. K. (1988). Eimeria tenella: Immunological diversity between asexual generations. Parasite Immunology 10, 649–60.CrossRefGoogle ScholarPubMed
Miller, G. A., Bhogal, B. S., McCandliss, R., Strausberg, R. L., Jessee, S. J., Anderson, A. C., Fuchs, C. K., Nagle, J., Likel, M. H., Strasser, J. M. & Strausberg, S. (1989). Characterisation and vaccine potential of a novel recombinant coccidial antigen. Infection and Immunity 57, 2014–20.Google Scholar
Ridley, R. G., Takacs, B., Lahm, U-W., Delves, C. J., Goman, M., Certa, U., Matile, H., Woollett, G. R. & Scaife, J. G. (1990). Characterisation and sequence of a protective rhoptry antigen from Plasmodium falciparum. Molecular and Biochemical Parasitology 41, 125–34.CrossRefGoogle ScholarPubMed
Sadak, A., Taghy, Z., Fortier, B. & Dubremetz, J-F. (1988). Characterization of a family of rhoptry proteins of Toxoplasma gondii. Molecular and Biochemical Parasitology 29, 203–12.CrossRefGoogle ScholarPubMed
Schamtz, D. M., Crane, M. ST. J. & Murray, P. K. (1984). Purification of Eimeria sporozoites by DE-52 anion exchange chromatography. Journal of Protozoology 31, 181–3.CrossRefGoogle Scholar
Speer, C. A., Thammnana, P. & Schenkel, R. H. (1989). Ultrastructural localization of antigenic sites on sporozoites, sporocysts, and oocysts of Eimeria acervulina and Eimeria tenella by monoclonal IgG and IgM antibodies. Journal of Parasitology 75, 92–7.CrossRefGoogle ScholarPubMed
Sutton, C. A., Shirley, M. W. & Wisher, M. H. (1989). Characterisation of coccidial proteins by two-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Parasitology 99, 175–87.CrossRefGoogle ScholarPubMed
Taylor, D. W., Evans, C. B., Aley, S. B., Barta, J. R. & Danforth, H. D. (1990). Identification of an apically-located antigen that is conserved in sporozoan parasites. Journal of Protozoology 37, 540–5.CrossRefGoogle ScholarPubMed
Tomley, F. M., Clarke, L. E., Kawazoe, U., Dijrema, R. & Kok, J. J. (1991). Sequence of the gene encoding an immunodominant microneme protein of Eimeria tenella. Molecular and Biochemical Parasitology (in the Press).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
Wisher, M. H. (1986). Identification of the sporozoite antigens of Eimeria tenella. Molecular and Biochemical Parasitology 21, 7–15.CrossRefGoogle ScholarPubMed