Localization of a Toxoplasma gondii Rhoptry Protein by Immunoelectron Microscopy During and After Host Cell Penetration |
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Authors: | LINDA D. SAFFER,ODILE MERCEREAU-PUIJALON,JEAN-FRANÇ OIS DUBREMETZ,JOSEPH D. SCHWARTZMAN |
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Affiliation: | Department of Pathology, University of Virginia School of Medicine, Charlottesville, Virginia 22908;Unitéde Parasitologie Experimental, Institut Pasteur, Paris, France 75015;INSERM Unité42, Villeneuve d'Ascq, France 59650 |
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Abstract: | ABSTRACT We immunolocalized a Toxoplasma gondii rhoptry protein (ROP1) before and after parasite host cell invasion of human fibroblasts and TG180 murine sarcoma cells by electron microscopy and immunogold labeling using either a monoclonal antibody (Tg49) or a monospecific rabbit antiserum (α249). At all stages of parasite growth ROP1 was found within the body but rarely within the peduncle of rhoptries, even in those that appeared empty. Immediately after host cell invasion ROP1 was associated with the parasitophorous vacuole membrane. Within hours after invasion the amount of ROP1 immunodetectable on the parasitophorous vacuole membrane was markedly decreased. The localization of ROP1 suggests a role in the early establishment of infection in host cells, consistent with previous work that has indicated that monoclonal antibodies to ROP1 (including the one used in these studies) interfere with the phenomenon of penetration enhancement. |
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Keywords: | Cultured cells electron microscopy human fibroblasts immune serum immunoblot immunolocalization techniques monoclonal antibody TG180 sarcoma cells |
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