Isolation and Characterization of Erythrocyte and Parasite Membranes from Rhesus Red Cells Infected with P. knowlesi.
Annual summary rept. no. 3, 1 Jun 78-31 May 79,
TUFTS-NEW ENGLAND MEDICAL CENTER BOSTON MA
Pagination or Media Count:
Intraerythrocytic maturation of P. knowlesi was achieved by cultivating infected cells in FEP-teflon containers, at controlled pO2, and pH, allowing , labeling of parasite components and parasite-induced glycoproteins in host-cell membranes with 14C-amino acids and 14C-sugars. Amino acid and glucosamine labeling were localized primarily in the parasite whereas 14C-galactose and 14C-fucose associated mostly with host cell membranes. Among host-cell membrane proteins preferential incorporation of 14C-amino acids and 14C-glucosamine occurred at pI 4.5-5.2 upon isoelectric focusing and these proteinsglycoproteins migrated between 90,000 and 45,000D upon SDS-PAGE. Monkey anti-schizont antisera and the serum of a monkey rendered naturally immune after infection with P. knowlesi showed the following immunoelectrophoretic results a twenty-four immune precipitates identified in purified schizonts. Seven components were common to schizonts and erythrocyte membranes, four unique to the infected membranes and the thirteen were exclusively found in the parasite. b Absorption of sera with schizont-infected erythrocytes eliminated at least three schizont immunoprecipitates and at least five host-cell membrane components. c CIE of 125I-labeled membrane proteins followed by autoradiography revealed that antigens 13 and 19 are exposed on the surfaces of infected cells. d Parasite-induced membrane components, 1 and 13, were immunogenic in vivo.
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