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Ultrastructural Localization of Protective Antigens of Plasmodium yoelii Merozoites by the Use of Monoclonal Antibodies and Ultrathin Cryomicrotomy
Mikio Oka
Mikio Oka
Institute of Pathology, Case Western Reserve University, Department of Molecular Biology, The Wellcome Research Laboratories, Cleveland, Ohio 44106, England
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Masamichi Aikawa
Masamichi Aikawa
Institute of Pathology, Case Western Reserve University, Department of Molecular Biology, The Wellcome Research Laboratories, Cleveland, Ohio 44106, England
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Robert R. Freeman
Robert R. Freeman
Institute of Pathology, Case Western Reserve University, Department of Molecular Biology, The Wellcome Research Laboratories, Cleveland, Ohio 44106, England
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Anthony A. Holder
Anthony A. Holder
Institute of Pathology, Case Western Reserve University, Department of Molecular Biology, The Wellcome Research Laboratories, Cleveland, Ohio 44106, England
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Edward Fine
Edward Fine
Institute of Pathology, Case Western Reserve University, Department of Molecular Biology, The Wellcome Research Laboratories, Cleveland, Ohio 44106, England
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The production of two hybridoma cell lines secreting monoclonal antibodies (MAb), both of which react specifically with erythrocytic merozoites of Plasmodium yoelii in the indirect immunofluorescence assay, has been reported earlier. MAb 25.77 was reactive with a localized region within each merozoite, while MAb 25.1 appeared to be specific for the plasma membrane of schizonts and merozoites. The parasite antigens recognized by antibodies 25.77 and 25.1 are proteins of 235,000 and 230,000 molecular weight, respectively, both of which induce protective immunity against P. yoelii in mice. In order to establish the precise localization of these protective antigens within erythrocyte merozoites, ultrathin cryomicrotomy was used in conjunction with the MAb and protein A-gold. This technique showed that gold particles were exclusively concentrated over the rhoptries when erythrocytic merozoites were incubated with MAb 25.77. On the other hand, gold particles were distributed uniformly over the merozoite surface when parasites were incubated with MAb 25.1. These results demonstrate, for the first time, that a protective antigen of the erythrocytic stage of P. yoelii is localized within the rhoptries as well as on the merozoite surface.
| Past two years | Past Year | Past 30 Days | |
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