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The development of exo-erythrocytic schizonts of Plasmodium berghei in vitro from gamma-irradiated and non-irradiated sporozoites: a study using confocal laser scanning microscopy

Published online by Cambridge University Press:  06 April 2009

R. E. Sinden
Affiliation:
Molecular and Cellular Parasitology Group, Department of Biology, Imperial College, London SW7 2BB
A. Couchman
Affiliation:
Molecular and Cellular Parasitology Group, Department of Biology, Imperial College, London SW7 2BB
A. Suhrbier
Affiliation:
Molecular and Cellular Parasitology Group, Department of Biology, Imperial College, London SW7 2BB
F. Marsh
Affiliation:
Molecular and Cellular Parasitology Group, Department of Biology, Imperial College, London SW7 2BB
L. Winger
Affiliation:
Molecular and Cellular Parasitology Group, Department of Biology, Imperial College, London SW7 2BB
G. Ranawaka
Affiliation:
Molecular and Cellular Parasitology Group, Department of Biology, Imperial College, London SW7 2BB

Extract

Confocal scanning laser microscopy has been used to study the distribution of antigens expressed by the liver stages of Plasmodium berghei in cultured hepatoma cells. The 3-dimensional images obtained of intact parasites clearly show complex patterns of antigen expression not apparent when using conventional IFAT or immunoelectron microscopy. A liver-stage specific antigen (Pbl 1) was shown to be confined to the parasitophorous vacuole; the vacuole has extensive diverticulae extending into the host cell. Small parasites were detected for the first time in ‘mature’ cultures. These did not represent a distinct population, but the ‘tail’ of a broad continuum of parasite sizes. Irradiated sporozoites produce a transient population of slow-growing parasites which express a very limited range of antigens de novo in the invaded hepatoma cell. A comparison of the reactivity of text-abstract EE parasites with anti-circumsporozoite antibody and with anti-Pbl 1 suggests that the former reagent may reliably be used to identify sporozoites invading host cells, but should not be used to determine the number of parasites that successfully undergo intrahepatic development. Anti-Pbl-1 indicates on 33% of invaded sporozoites identified by anti-CSP subsequently differentiate.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1991

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References

Bianco, A. E., Crewther, P. E., Coppel, R. L., Stahl, H. D., Kemp, D. J., Anders, R. F. & Brown, G. V. (1988). Patterns of antigen expression in asexual stages and gametocytes of Plasmodium falciparum. American Journal of Tropical Medicine and Hygiene 38, 258–67.Google Scholar
Hollingdale, M. R., Nardin, E. H., Tharavanij, S., Schwartz, A. L. & Nussenzweig, R. S. (1984). Inhibition of entry of Plasmodium falciparum and P. vivax sporozoites into cultured cells: an in vitro assay of protective antibodies. Journal of Immunology 132, 909–13.CrossRefGoogle Scholar
Krotoski, W. A., Krotoski, D. M., Garnham, P. C. C., Bray, R. S., Killick-Kendrick, R., Draper, C. C., Targett, G. A. T. & Guy, M. W. (1980). Relapses in primate malaria. Discovery of two populations of exoerythrocytic stages. A preliminary note. British Medical Journal 1, 153.Google Scholar
Landau, I. & Killick-Kendrick, R. (1966). Rodent plasmodia of the République Centrafricaine: the sporogony and tissue stages of Plasmodium chabaudi and P. berghei yoelii. Transactions of the Royal Society of Tropical Medicine and Hygiene 60, 633–49.CrossRefGoogle ScholarPubMed
Meis, J. F. G. M., Verhave, J. P., Jap, P. H. K., Sinden, R. E. & Meuwissen, J. H. E. T. (1983). Malaria parasites: discovery of the early liver form. Nature, London 302, 424–6.CrossRefGoogle ScholarPubMed
Mellouk, S., Lunel, F., Sedegah, M., Beaudoin, R. L. & Druilhe, P. (1990). Protection against malaria induced by irradiated sporozoites. Lancet 335, 721.CrossRefGoogle ScholarPubMed
Nussenzweig, R. S., Vanderberg, J. P., Most, H. & Orton, C. (1967). Protective immunity produced by injection of X-irradiated sporozoites of Plasmodium berghei. Nature, London 216, 160–2.Google Scholar
Renia, L., Miltgen, F., Charoenvit, Y., Ponnudurai, T., Verhave, J. P., Collins, W. E. & Mazier, D. (1988). Malaria sporozoite penetration: a new approach by double staining. Journal of Immunological Methods 112, 201–5.Google Scholar
Shortt, H. E. & Garnham, P. C. C. (1948). Demonstration of a persisting exo-erythrocytic cycle in P. cynomolgi and its bearing on the production of relapse. British Medical Journal 1, 1225–8.CrossRefGoogle Scholar
Sinden, R. E., Suhrbier, A., Davies, C., Fleck, S., Hodivala, K. & Nicholas, J. C. (1990). The development and routine application of high-density exoerythrocytic-stage cultures of Plasmodium berghei. Bulletin of the World Health Organization (in the Press),Google Scholar
Suhrbier, A., Janse, C. J., Mons, B., Fleck, S. L., Nicholas, J., Davies, C. S. & Sinden, R. E. (1987). The complete development in vitro of the vertebrate stages of the rodent malaria parasite Plasmodium berghei. Transactions of the Royal Society of Tropical Medicine and Hygiene 81, 907–10.Google Scholar
Suhrbier, A., Hamilton, A. J., Nicholas, J. & Sinden, R. E. (1988). The fate of the circumsporozoite antigen during the exoerythrocytic stage of Plasmodium berghei. European Journal of Cell Biology 46, 2530.Google Scholar
Suhrbier, A., Castellano, E. & Sinden, R. E. (1990 a). The survival and antigenic profile of the irradiated sporozoite in the infected liver cell. Infection and Immunity 58, 2834–9.CrossRefGoogle Scholar
Suhrbier, A., Winger, L. A., O'dowd, C. A., Hodivala, K. J. & Sinden, R. E. (1990 b). An antigen specific to the liver stage of rodent malaria recognised by a monoclonal antibody. Parasite Immunology 12, 473–81.CrossRefGoogle Scholar
Winger, L. A., Sinden, R. E., O'dowd, C. A., Hodivala, K. J. & Suhrbier, A. (1990). A liver-stage specific antigen of Plasmodium berghei identified by a monoclonal antibody. Bulletin of the World Health Organization (in the Press),Google ScholarPubMed