Monoclonal Antibodies Identify a Subset of Dense Granules in Cryptosporidium parvum Zoites and Gamonts |
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Authors: | ALAIN BONNIN JIRI GUT JEAN FRANCOIS DUBREMETZ RICHARD G. NELSON PATRICK CAMERLYNCK |
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Affiliation: | Laboratoire de Parasitologie-Mycologie, Hôpital du Bocage, Boulevard de Lattre de Tassigny, 21034 Dijon Cedex, France;Parasitology Laboratory, Division of Infectious Diseases, San Francisco General Hospital and Departments of Medicine and Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143–0811, USA;INSERM U 42, Domaine du Certia, 369 rue Jules Guesde, 59650 Villeneuve D'Ascq, France |
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Abstract: | ABSTRACT. Two monoclonal antibodies raised against purified oocysts and excysted sporozoites of Cryptosporidium parvum identified antigens located in the anterior half of sporozoites by indirect immunofluorescence microscopic assay. The monoclonal antibodies also reacted with Triton X-100-insoluble antigens of asexual and sexual stage parasites developing in epithelial cells in vitro and identified a 110 kilodalton antigen on immunoblots of sodium dodecyl sulfate-extracted oocysts. Immunoblotting reactivity was abolished by prior treatment of blotted antigen with periodic acid suggesting that the monoclonal antibodies recognize a carbohydrate or carbohydrate-dependent epitope(s). By immunoelectron microscopy, the antibodies reacted with a family of small, electron-dense granules located predominantly in the central region of merozoites and also with a population of cytoplasmic inclusions in macrogamonts. In addition, the monoclonal antibodies prominently labeled the parasitophorous vacuole membrane of all intracellular stages examined suggesting that the corresponding antigen(s) may be exocytosed from the granules to become associated with Triton X-100-insoluble components of the vacuolar membrane or cytoskeleton. |
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Keywords: | Apical complex Apicomplexa Coccidia exocytosis host cell invasion merozoite parasitophorous vacuole sporozoite vacuolar wall |
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