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Plasmodium falciparum: isolation of intact and erythrocyte-free trophozoites from sorbitol lysates

Published online by Cambridge University Press:  06 April 2009

H. C. Hoppe
Affiliation:
Department of Biochemistry
J. Coetzee
Affiliation:
Electron Microscopy Unit, University of Pretoria, Pretoria 0002, Republic of South Africa
A. I. Louw
Affiliation:
Department of Biochemistry

Extract

An in vitro culture of Plasmodium falciparum (isolate FCUP-1/RSA) was arrested in the trophozoite stage with α-difluoromethylornithine (DFMO), followed by sorbitol treatment to liberate intracellular parasites from infected erythrocytes. Most of the unlysed erythrocytes and ghost membranes were removed by filtration, after agglutination with anti-erythrocyte monoclonal antibodies or wheat-germ agglutinin. A highly purified parasite preparation was obtained after chromatography on an immuno-affinity column consisting of polystyrene particles to which anti-erythrocyte monoclonal antibodies had been adsorbed. Isolated parasites were free of surrounding erythrocyte membranes and structurally intact as assessed by transmission electron microscopy, SDS–PAGE and ELISA.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1992

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References

Aley, S. B., Sherwood, J. A., Marsh, K., Eidelman, O. & Howard, R. J. (1986). Identification of isolate-specific proteins on sorbitol-enriched Plasmodium falciparum-infected erythrocytes from Gambian patients. Parasitology 92, 511–25.Google Scholar
Assaraf, Y. G., Golenser, J., Spira, D. T. & Bachrach, U. (1986). Plasmodium falciparum: synchronisation of cultures with D, L-α-difluoromethylornithine, an inhibitor of polyamine biosynthesis. Experimental Parasitology 61, 229–35.Google Scholar
Banyal, H. S. & Inselburg, J. (1985). Isolation and characterization of parasite-inhibitory Plasmodium falciparum monoclonal antibodies. American Journal of Tropical Medicine and Hygiene 34, 1055–64.CrossRefGoogle ScholarPubMed
Beaumelle, B. D., Vial, H. J. & Philippot, J. R. (1987). Re-evaluation, using marker enzymes, of the ability of saponin and ammonium chloride to free Plasmodium from infected erythrocytes. Journal of Parasitology 73, 743–8.Google Scholar
Butler, J. E., Peterman, J. H., Suter, M. & Dierks, S. E. (1987). The immunochemistry of solid-phase sandwich enzyme-linked immunosorbent assays. Federation Proceedings 46, 2548–56.Google Scholar
Conradie, J. D., Govender, M. & Visser, L. (1983). ELISA solid-phase: partial denaturation of coating antibody yields a more efficient solid phase. Journal of Immunological Methods 59, 289–99.Google Scholar
Cook, R. T., Aikawa, M., Rock, R. C., Little, W. & Sprinz, H. (1969). The isolation and fractionation of Plasmodium knowlesi. Military Medicine, Special Issue, 1969, 866–83.Google Scholar
GALFRÉ, G. & Milstein, C. (1981). Preparation of monoclonal antibodies: strategies and procedures. Methods in Enzymology 73, 346.Google Scholar
Ginsburg, H., Kutner, S., Zangwil, M. & Cabantchik, Z. I. (1986). Selectivity properties of pores induced in host erythrocyte membranes by Plasmodium falciparum. Effect of parasite maturation. Biochimica et Biophysica Acta 861, 194–6.Google Scholar
Hamburger, J. & Kreier, J. P. (1980). Isolation of malarial parasites and their constituents. In Malaria Vol. 3. Immunology and Immunization, pp. 165. New York: Academic Press.Google Scholar
Heidrich, H.-G., Mrema, J. E. K., Van DerJagt, D. L. Jagt, D. L., Reyes, P. & Rieckmann, K. H. (1982). Isolation of intracellular parasites (Plasmodium falciparum) from culture using free-flow electrophoresis: separation of the free parasites according to stages. Journal of Parasitology 68, 443–50.Google Scholar
Hoppe, H. C., Verschoor, J. A. & Louw, A. I. (1991). Plasmodium falciparum: a comparison of synchronisation methods for in vitro cultures. Experimental Parasitology 72, 464–7.Google Scholar
Kara, U. A. K., Stenzel, D. J., Ingram, L. T. & Kidson, C. (1988). The parasitophorous vacuole membrane of Plasmodium falciparum: demonstration of vesicle formation using an immunoprobe. European Journal of Cell Biology 46, 917.Google Scholar
Killby, V. A. A. & Silverman, P. H. (1969). Isolated erythrocytic forms of Plasmodium berghei. American Journal of Tropical Medicine and Hygiene 18, 836–59.Google Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.Google Scholar
Lambros, C. & Vandenberg, J. P. (1979). Synchronisation of Plasmodium falciparum erythrocyte stages in culture. Journal of Parasitology 65, 418–20.Google Scholar
Langreth, S. G., Jensen, J. B., Reese, R. T. & Trager, W. (1978). Fine structure of human malaria. Journal of Protozoology 25, 443–52.Google Scholar
Lee, M. & Lambros, C. (1988). The ELISA-U: an enzyme-linked immunosorbent assay using urease as the enzyme marker for rapid detection of Plasmodium falciparum antibody in human serum. American Journal of Tropical Medicine and Hygiene 39, 421–6.Google Scholar
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265–75.Google Scholar
Mikkelsen, R. B., Singhal, A. & SCHMIDT-Ullrich, R. (1989). Erythrocyte membrane proteins in the parasitophorous vacuole membrane of Plasmodium falciparum. Journal of Cell Biochemistry 13E (Suppl.), 112.Google Scholar
Mrema, J. E. K., Langreth, S. G., Jost, R. C., Rieckmann, K. H. & Heidrich, H.-G. (1982). Plasmodium falciparum: isolation and purification of spontaneously released merozoites by nylon membrane sieves. Experimental Parasitology 54, 285–95.Google Scholar
Nillni, E. A., SCHMIDT-Ullrich, R., Mikkelsen, R. B. & Wallach, D. F. H. (1985). Extracellular development of Plasmodium knowlesi erythrocytic stages in an artificial intracellular medium. Molecular and Biochemical Parasitology 17, 219–38.Google Scholar
O'Farrel, P. H. (1974). High resolution two-dimensional electrophoresis of proteins. Journal of Biological Chemistry 250, 4007–21.Google Scholar
Read, L. K. & Mikkelsen, R. B. (1991). Plasmodium falciparum infected erythrocytes contain an adenylate cyclase with properties which differ from those of the host enzyme. Molecular and Biochemical Parasitology 45, 109–20.Google Scholar
Severs, N. J. & Robenek, H. (1983). Detection of microdomains in biomembranes. An appraisal of recent developments in freeze-fracture cytochemistry. Biochimica et Biophysica Acta 737, 373408.Google Scholar
Siddiqui, W. A. (1977). An effective immunisation of experimental monkeys against a human malaria parasite, Plasmodium falciparum. Science 197, 388–9.Google Scholar
Sterling, C. R. (1984). In vitro isolation of Plasmodium chabaudi merozoites by continuous flow ultrasound, cell sieving, concanavalin A-affinity chromatography and poly-L-lysine coated bead support columns. Journal of Parasitology 70, 945–54.Google Scholar
Trager, W. & Jensen, J. B. (1976). Human malaria parasites in continuous culture. Science 193, 673–5.Google Scholar
Trager, W., Langreth, S. G. & Platzer, E. G. (1972). Viability and fine structure of extracellular Plasmodium lophurae prepared by different methods. Proceedings of the Helminthological Society of Washington 39, 220–30.Google Scholar
Trager, W., Zung, J. & Tershakovec, M. (1990). Initial extracellular development in vitro of erythrocytic stages of malaria parasites (Plasmodium falciparum). Proceedings of the National Academy of Sciences, USA 87, 5618–22.Google Scholar
Vernes, A., Haynes, J. D., Tapchaisri, P., Williams, J. L., Dutoit, E. & Diggs, C. L. (1984). Plasmodium falciparum strain-specific human antibody inhibits merozoite invasion of erythrocytes. American Journal of Tropical Medicine and Hygiene 33, 197203.Google Scholar
Verschoor, J. A., Coetzee, L. & Visser, L. (1989). Monoclonal antibody characterisation of two field strains of Haemophilus paragallinarum isolated from vaccinated layer hens. Avian Diseases 33, 219–25.CrossRefGoogle ScholarPubMed
Wunderlich, F., Helwig, M., Schillinger, G., Vial, H., Philippot, J. & Speth, V. (1987). Isolation and characterization of parasites and host cell ghosts from erythrocytes infected with Plasmodium chabaudi. Molecular and Biochemical Parasitology 23, 103–15.Google Scholar
Wunderlich, F., Schillinger, G. & Helwig, M. (1985). Fractionation of Plasmodium chabaudi-infected erythrocytes into parasites and ghosts. Zeitschrift für Parasitenkunde 71, 545–51.Google Scholar