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Characterization and immunolocalization of an oocyst wall antigen of Cryptosporidium parvum (Protozoa: Apicomplexa)

Published online by Cambridge University Press:  06 April 2009

A. Bonnin
Affiliation:
Laboratoire de Parasitologie-Mycologie, Hôpital du Bocage, 21034 Dijon Cedex, France
J. F. Dubremetz
Affiliation:
INSERM U42, Domaine du Certia, 59650 Villeneuve d'Ascq, France
P. Camerlynck
Affiliation:
Laboratoire de Parasitologie-Mycologie, Hôpital du Bocage, 21034 Dijon Cedex, France

Abstract

A monoclonal antibody (OW-IGO) raised against purified excysted oocysts of Cryptosporidium parvum reacted in an immunofluorescence assay with the oocyst wall. The corresponding antigen was localized by immunoelectron microscopy in fibrillous material present in the parasitophorous vacuole of developing macrogametes and in the wall of both single and double layered sporulating oocysts. Gold particles were also detected over electron-lucent vesicles of the macrogametes by immunoelectron microscopy. On Western blotting of C. parvum oocyst extracts, major bands at 250 and 40 kDa and several minor components were recognized by Mab OW-IGO. Almost complete abolition of Western blot reactivity occurred after periodate oxidation of oocyst antigen, suggesting that monoclonal antibody OW-IGO reacts with a carbohydrate epitope. Taken together, our results suggest that a fibrillous glycoproteinic material is released in the parasitophorous vacuole from electron-lucent vesicles during gametogenesis, and later condensed in the oocyst wall.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1991

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References

Anusz, K. Z., Mason, P. H., Riggs, M. W. & Perryman, L. E. (1990). Detection of Cryptosporidium parvum oocysts in bovine feces by monoclonal antibody capture enzyme-linked immunosorbent assay. Journal of Clinical Microbiology 28, 2770–4.CrossRefGoogle ScholarPubMed
Bonnin, A., Dubremetz, J. F. & Camerlynck, P. (1991). Characterization of microneme antigens of Cryptosporidium parvum (Protozoa, Apicomplexa). Infection and Immunity 59, 1703–8.CrossRefGoogle ScholarPubMed
Bonnin, A., Salimbeni, I., Dubremetz, J. F., Harly, G., Chavanet, P. & Camerlynck, P. (1990 a). Mise au point d'un modèle expérimental de culture in vitro des stades asexués de Cryptosporidium sp. Annales de Parasitologie Humaine et Comparée 65, 41–3.CrossRefGoogle Scholar
Bonnin, A., Petrella, T., Dubremetz, J. F., Michiels, J. F., Puygauthier-Toubas, D. & Camerlynck, P. (1990 b). Histopathological method for diagnosis of cryptosporidiosis using monoclonal antibodies. European Journal of Clinical Microbiology and Infectious Diseases 9, 664–6.CrossRefGoogle ScholarPubMed
Campbell, I., Tzipori, S., Hutchison, G. & Angus, K. W. (1982). Effect of disinfectants on survival of Cryptosporidium oocysts. Veterinary Record 111, 414–15.CrossRefGoogle ScholarPubMed
Current, W. L. & Reese, N. C. (1986). A comparison of endogenous development of three isolates of Cryptosporidium in suckling mice. Journal of Protozoology 33, 98108.CrossRefGoogle ScholarPubMed
Doens-Juteau, O. & Senaud, J. (1974). Etude ultrastructurale de la paroi du macrogamète et de l'oocyste chez Eimeria tenella (Railliet et Lucet, 1891), coccidie Eimeriidae, parasite de l'intestin des poulets. Protistologica 10, 261–70.Google Scholar
Dubremetz, J. F. & Yvore, P. (1971). Elaboration de la coque oocystique chez la Coccidie Eimeria necatrix Johnson 1930 (Sporozoaires, Coccidiomorphes). Etude au microscope électronique. Comptes rendus des Séances de la Société de Biologie 165, 862.Google Scholar
Ferguson, D. F. P., Hutchison, W. M. & Siim, J. C. (1975). The ultrastructural development of the macrogamete and formation of the oocyst wall of Toxplasma gondii. Acta Pathologica Microbiologica Scandinavica, Section B 83, 491505.Google Scholar
Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Luft, B. J., Payne, D., Woodmansee, D. & Kim, C. W. (1987). Characterization of the Cryptosporidium antigens from sporulated oocysts of Cryptosporidium parvum. Infection and Immunity 55, 2436–41.CrossRefGoogle ScholarPubMed
Rehg, J. E., Hancock, M. L. & Woodmansee, D. B. (1987). Characterization of cyclophosphamide-rat model of Cryptosporidiosis. Infection and Immunity 55, 2669–74.CrossRefGoogle ScholarPubMed
Smith, H. V. & Rose, J. B. (1990). Waterborne cryptosporidiosis. Parasitology Today 6, 812.CrossRefGoogle ScholarPubMed
Stotish, R. L., Wang, C. C. & Meyenhofer, M. (1978). Structure and composition of the oocyst wall of Eimeria tenella. Journal of Parasitology 64, 1074–81.CrossRefGoogle ScholarPubMed
Ungar, B. L. P., Soave, R., Fayer, R. & Nash, T. E. (1986). Enzyme immunoassay detection of immunoglobulin M and G antibodies to Cryptosporidium in immunocompetent and immunocompromised persons. Journal of Infectious Diseases 153, 570–8.CrossRefGoogle Scholar
Vetterling, J. M., Pacheco, N. D. & Fayer, R. (1973). Fine structure of gametogony and oocyst formation in Sarcocystis sp. in cell culture. Journal of Protozoology 20, 613–21.CrossRefGoogle ScholarPubMed
Wang, C. C. (1982). Biochemistry and physiology of Coccidia. In The Biology of the Coccidia (ed. Long, P. L.) pp. 167216. Baltimore: University Park Press.Google Scholar
Woodward, M. P., Young, W. W. Jr & Bloodgood, R. A. (1985). Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. Journal of Immunological Methods 78, 143–53.CrossRefGoogle ScholarPubMed