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Localization of parasite antigens in Cryptosporidium parvum-infected epithelial cells using monoclonal antibodies

Published online by Cambridge University Press:  06 April 2009

V. McDonald ,
M. V. McCrossan  and
F. Petry
V. McDonald
Affiliation:
Department of Clinical Sciences, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT
M. V. McCrossan
Affiliation:
Department of Medical Parasitology, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT
F. Petry
Affiliation:
Department of Medical Parasitology, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT
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Summary

An immunogold ultrastructural study was made of Cryptosporidium parvum-infected intestinal cells from SCID mice to locate parasite antigens recognized by monoclonal antibodies raised against sporozoite or oocyst wall antigens. The results suggested that these antigens were present in more than one life-cycle stage and demonstrated that the intracellular parasite modified the parasitophorous vacuole membrane and villous membrane surrounding the parasite. In an immuno-fluorescence antibody test monoclonal antibody (MAb) IB5 reacted with the oocyst wall, MAb 2C3 with the whole sporozoite and MAb 2B2 with the sporozoite surface. Western and dot-blot studies demonstrated that different carbohydrate epitopes were recognized by the respective sporozoite-reactive antibodies. In the ultrastructural examination MAb 1B5 reacted with macro- and microgametocytes as well as the oocyst wall. In the macrogametocyte MAb 1B5 recognized the large electron-dense bodies characteristic of this stage and, in some parasites, the parasitophorous vacuole and the parasite pellicle. The sporozoite-reactive MAbs were able to bind to all developmental stages. These antibodies recognized the parasite cytoplasm and, additionally, MAb 2B2 produced substantial labelling of the parasite membrane. Significantly, both these antibodies also detected antigen in the parasitophorous vacuole membrane and, to a lesser extent, the villous membrane surrounding the parasite.

Keywords

Cryptosporidium parvummonoclonal antibodyWestern blottingimmunogold labellingelectron microscopy.
Type
Research Article
Information
Parasitology , Volume 110 , Issue 3 , April 1995 , pp. 259 - 268
Copyright
Copyright © Cambridge University Press 1995

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References

REFERENCES

Bjorneby, J. M., Riggs, M. W. & Perrymann, L. E. (1990) Cryptosporidium parvum merozoites share neutralization-sensitive epitopes with sporozoites. Journal of Immunology 145, 298–304.Google Scholar
Blake, M. S., Johnston, K. H., Russel-Johes, G. J. & Gotschlich, E. C. (1984) A rapid, sensitive method for detection of alkaline phosphatase-conjugated antiantibody on Western blots. Analytical Biochemistry 136, 175–9.Google Scholar
Bonnin, A., Dubremetz, J. F. & Camerlynck, P. (1991) Characterization and immunolocalization of an oocyst wall antigen of Cryptosporidium parvum (Protozoa: Apicomplexa). Parasitology 103, 171–7.Google Scholar
Current, W. L. & Garcia, L. S. (1991) Cryptosporidiosis. Clinical Microbiology Reviews 4, 325–58.Google Scholar
Current, W. L. & Reese, N. C. (1986) A comparison of endogenous development of three isolates of Cryptosporidium in suckling mice. Journal ofProtozoology 33, 98–108.Google Scholar
Dubremetz, J. F. (1993) Apical organelles (rhoptries, micronemes, dense granules) and host cell invasion by Coccidia: What do we know now? In Proceedings of the VIth International Coccidiosis Conference, June 21–25, 1993 (ed. Barta, J. R. & Fernando, M. A.), pp. 3–9. University of Guelph, Ontario.Google Scholar
Elmendorf, H. G. & Haldar, K. (1993) Secretory transport in Plasmodium. Parasitology Today 9, 98–102.Google Scholar
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.Google Scholar
Lazo, A., Barriga, O. O., Redman, D. R. & Bech-Nielsen, s. (1986) Identification by transfer blot of antigens reactive with the enzyme-linked immunosorbent assay (ELISA) in rabbits immunized and calf infected with Cryptosporidium sp. Veterinary Parasitology 21, 151–63.Google Scholar
Lumb, R., Lanser, J. A. & O'Donoghue, P. J. (1988) Electrophoretic and immunoblot analysis of Cryptosporidium. Immunology and Cell Biology 66, 369–76.Google Scholar
Lumb, R., Smith, P. S., Davies, R., O'Donoghue, P. J., Atkinson, H. M. & Lanser, J. A. (1989) Localization of a 23 000 MW antigen of Cryptosporidium by immunoelectron microscopy. Immunology and Cell Biology 67, 267–70.Google Scholar
Marcial, M. A. & Madara, J. L. (1986) Cryptosporidium: cellular localization, structural analysis of absorptive cell—parasite membrane—membrane interactions in guinea-pigs, and suggestion of protozoal transport by M cells. Gastroenterology 90, 583–94.CrossRefGoogle Scholar
McDonald, v. & Bancroft, G. J. (1994) Mechanisms of innate and acquired resistance to Cryptosporidium parvum infection in SCID mice. Parasite Immunology 16, 315–20.CrossRefGoogle ScholarPubMed
McDonald, v., Deer, R. M. A., Nina, J. M. S., Wright, s., Chiodini, P. L. & Mcadam, K. P. W. J. (1991) Characteristics and specificity of hybridoma antibodies against oocyst antigens of Cryptosporidium parvum. Parasite Immunology 13, 251–9.CrossRefGoogle ScholarPubMed
McDonald, V., Stables, R., Warhurst, D. C., Barer, M. R., Blewett, D. A., Chapman, H. D., Connolly, G. M., Chiodini, P. L. & McAdam, K. P. W. J. (1990) In vitro cultivation of Cryptosporidium parvum and screening for anticryptosporidial drugs. Antimicrobial Agents and Chemotherapy 34, 1498–500.Google Scholar
Nina, J. M. S., McDonald, v., Dyson, D. A., Catchpole, J., Uni, S., Iseki, M., Chiodini, P. L. & McAdam, K. P. W. J. (1992) Analysis of oocyst wall and sporozoite antigens from three Cryptosporidium species. Infection and Immunity 60, 1509–13.CrossRefGoogle Scholar
Robert, B., Ginter, A., Antoine, H., Coppe, P. & Collard, A. (1989) Cryptosporidium parvum: immunolabelling with monoclonal antibodies. In Coccidia and Intestinal Coccidiomorphs, Vth International Coccidiosis Conference (ed. Yvorre, P.), p. 23. I.N.R.A. Publications, France.Google Scholar
Roberts, D. J., Biggs, B.-A., Brown, G. & Newbold, C. I. (1993) Protection, pathogenesis and phenotypic plasticity in Plasmodium falciparum malaria. Parasitology Today 9, 281–6.Google Scholar
Scholtyseck, E., Rommel, A. & Heller, G. (1969) Licht-und elektronmikroskopische Untersuchungen zur Bildung der Oocystenhülle bei Eimerien (Eimeria perforans, E. stiedae und E. tenella). Zeitschrift für Parasitenkunde 31, 289–98.Google Scholar
Stroncek, D. F., Skubitz, K. M. & McCullogh, J. J. (1990) Biochemical characterization of the neutrophil-specific antigen NB1. Blood 75, 744–55.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.Google Scholar
Tyzzer, E. E. (1907) A sporozoan found in the peptic glands of the common mouse. Proceedings of the Society for Experimental Biology and Medicine 5, 12–13.Google Scholar
Ungar, B. L. P., Burris, J. A., Quinn, C. A. & Finkelman, F. D. (1990) New mouse models for chronic Cryptosporidium infection in immunodeficient hosts. Infection and Immunity 58, 961–9.Google Scholar
Vetterling, J. M., Takeuchi, A. & Madden, P. A. (1971) Ultrastructure of Cryptosporidium wrairi from the guinea-pig. Journal of Protozoology 18, 248–60.CrossRefGoogle ScholarPubMed