Isolation and characterization of parasite-inhibitory Plasmodium falciparum monoclonal antibodies.
Three mouse hybridomas were isolated that produced IgM monoclonal antibodies (Mab) which reacted with erythrocytic stages of Plasmodium falciparum and inhibited the invasion of erythrocytes in vitro. Those Mab, initially identified by an ELISA screening of hybridoma culture medium, exhibited a strong binding to trophozoite and schizont but not to ring or merozoite stage parasites or to erythrocytes in an indirect immunofluorescence assay. All inhibited the parasite's ability to infect erythrocytes in an in vitro invasion inhibition assay. Western blot analysis of the binding of the Mab to SDS-PAGE-separated parasite antigens isolated from the ring, trophozoite, schizont or spontaneously released merozoite stages, indicated that two of the Mab bound to a Mr 105,000 antigen in trophozoites and schizonts while the third Mab did not. All three Mab also bound to Mr 30,000-40,000 antigens in all stages, however, in all instances binding to these antigens was enhanced in merozoites. It was further observed that the two Mabs that bound to a Mr 105,000 antigen: exhibited a markedly reduced binding to the Mr 105,000 antigen in merozoite preparations; exhibited different relative intensities of binding to the trophozoite and schizont antigens; both bound to the same Mr 105,000 antigen as demonstrated through Western blot analysis of antigens separated by two-dimensional gel electrophoresis. The findings suggest that the inhibitory Mab bound to different epitopes of the same antigen and that the antigen may either be processed or degraded at about the time of merozoite release and erythrocyte invasion.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D000953:Antigens, Protozoan / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000469:parasitology; D005455:Fluorescent Antibody Technique; D006825:Hybridomas; D007075:Immunoglobulin M / Q000276:immunology; D051379:Mice; D010963:Plasmodium falciparum / Q000254:growth & development / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D007075:Immunoglobulin M