Study of the antigenicity of P. yoelii parasitized erythrocyte ghost antigens and their role in protection

Terrientes S., Zilka I.
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The overall objective of this dissertation was the study of membrane antigens of parasitized erythrocytes in Plasmodium yoelii, a rodent malaria model. Unstimulated peritoneal cells from C57/Bl 10 mice phagocytosed P. yoelii parasitized erythrocytes in vitro more effectively in the presence of immune and hyperimmune sera than normal sera, suggesting these antibodies react with novel antigens expressed in the membrane of parasitized erythrocytes. A surface immunofluorescence assay used to detect the binding of hyperimmune serum antibodies to surface antigens was suggestive of, but did not conclusively demonstrate, the presence of novel surface antigens on parasitized erythrocytes. It was not possible to measure specific anti-plasmodial antibody using an enzyme linked immonosorbent assay (ELISA) although different variations of the technique were pursued. There was no significant difference between the binding of normal or hyperimmune sera to normal erythrocyte ghosts or R. yoelii parasitized erythrocyte ghosts in the ELISA. More conclusive evidence for the presence of parasite-derived membrane antigens in infected erythrocytes were obtained by 50S-polyacrylamide gel electrophoresis, immunoblotting and immunoprecipitation of biosynthetically labeled membrane antigens. Plasma membranes of P. yoelii trophozoites contained ten neoproteins with apparent molecular masses of 92.5, 79, 76, 69, 60, 43, 31, 30, 19 and 14.3 kDa. These proteins were metabolically labeled with 3H Isoleucine and immunoprecipitated by hyperimmune serum, however not all of these proteins were detected by immunoblotting. Some of these proteins appeared to have co-migrating normal host membrane antigens (92.5, 80 and 69 kDa). A vaccination experiment was performed to determine whether parasitized membrane preparations could induce an immune response capable of protecting mice against P. yoelii infection. BALB/c mice immunized intravenously with a single dose of P. yoelii parasitized erythrocyte ghosts without adjuvant developed a significant level of protective immunity against challenge with the homologous parasite as compared to controls, immunized with normal erythrocyte ghosts. In contrast, mice vaccinated with either two doses of normal erythrocyte ghosts or P. yoelii parasitized erythrocyte ghosts with Complete Freund's Adjuvant injected subcutaneously into the tail were not protected against challenge with P. yoelii parasites. The results of this study support a possible role of parasitized erythrocyte membrane antigens in immunity to malaria in the P. yoelii mouse malaria model.
Thesis (Ph. D.)--University of Hawaii at Manoa, 1990.
Includes bibliographical references (leaves 134-152)
xvi, 152 leaves, bound ill. 29 cm
Plasmodium yoelii, Antigen-antibody reactions, Erythrocyte disorders, Malaria -- Immunological aspects
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