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. 1992 Jan;60(1):264–270. doi: 10.1128/iai.60.1.264-270.1992

Early hepatic stages of Plasmodium berghei: release of circumsporozoite protein and host cellular inflammatory response.

Z M Khan 1, C Ng 1, J P Vanderberg 1
PMCID: PMC257531  PMID: 1729189

Abstract

After injection of Plasmodium berghei sporozoites into Norway-Brown rats, we were able to localize these sporozoites and the early hepatic trophozoites developing from them in histological sections of the liver stained with a sensitive immunogold-silver procedure. Sporozoites invading hepatocytes released substantial quantities of circumsporozoite protein into the hepatocyte cytoplasm. This intrahepatic cytoplasmic distribution reached a maximal level at about 4 h post-sporozoite injection. As the hepatic parasites continued to differentiate, circumsporozoite protein became undetectable within the cytoplasm of the hepatocytes and became localized around the periphery of each parasite. There was generalized cellular inflammation within the liver of the host, which first became evident at around 4 h post-sporozoite injection and progressed to the formation of well-defined granulomas by 24 h. Such histopathological changes were not seen in rats injected with killed sporozoites, indicating that the cellular inflammation was induced by viable, infective sporozoites. We did not observe cellular infiltration specifically associated with any of the developing hepatic stages that we observed, even up to 28 h post-sporozoite inoculation. These results indicate that viable sporozoites induced rapid and generalized hepatic inflammation in host rats. However, sporozoites that successfully invaded hepatocytes and then proceeded to develop further did not appear to be the target of inflammatory cells until a period beginning at around 40 h post-sporozoite inoculation.

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Selected References

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  1. Aikawa M., Schwartz A., Uni S., Nussenzweig R., Hollingdale M. Ultrastructure of in vitro cultured exoerythrocytic stage of Plasmodium berghei in a hepatoma cell line. Am J Trop Med Hyg. 1984 Sep;33(5):792–799. doi: 10.4269/ajtmh.1984.33.792. [DOI] [PubMed] [Google Scholar]
  2. Aley S. B., Atkinson C. T., Aikawa M., Maloy W. L., Hollingdale M. R. Ultrastructural localization of Plasmodium falciparum circumsporozoite protein in newly invaded hepatoma cells. J Parasitol. 1987 Dec;73(6):1241–1245. [PubMed] [Google Scholar]
  3. Atkinson C. T., Aikawa M., Aley S. B., Hollingdale M. R. Expression of Plasmodium berghei circumsporozoite antigen on the surface of exoerythrocytic schizonts and merozoites. Am J Trop Med Hyg. 1989 Jul;41(1):9–17. [PubMed] [Google Scholar]
  4. Danforth H. D., Orjih A. U., Nussenzweig R. S. Immunofluorescent staining of exoerythrocytic schizonts of Plasmodium berghei in fixed liver tissue with stage-specific immune serum. J Parasitol. 1978 Dec;64(6):1123–1125. [PubMed] [Google Scholar]
  5. De Waele M., Renmans W., Segers E., Jochmans K., Van Camp B. Sensitive detection of immunogold-silver staining with darkfield and epi-polarization microscopy. J Histochem Cytochem. 1988 Jun;36(6):679–683. doi: 10.1177/36.6.3259250. [DOI] [PubMed] [Google Scholar]
  6. Hamilton A. J., Suhrbier A., Nicholas J., Sinden R. E. Immunoelectron microscopic localization of circumsporozoite antigen in the differentiating exoerythrocytic trophozoite of Plasmodium berghei. Cell Biol Int Rep. 1988 Feb;12(2):123–129. doi: 10.1016/0309-1651(88)90126-9. [DOI] [PubMed] [Google Scholar]
  7. Hollingdale M. R., Leland P., Leef J. L., Leef M. F., Beaudoin R. L. Serological reactivity of in vitro cultured exoerythrocytic stages of Plasmodium berghei in indirect immunofluorescent or immunoperoxidase antibody tests. Am J Trop Med Hyg. 1983 Jan;32(1):24–30. doi: 10.4269/ajtmh.1983.32.24. [DOI] [PubMed] [Google Scholar]
  8. Hollingdale M. R., Leland P., Leef J. L., Schwartz A. L. Entry of Plasmodium berghei sporozoites into cultured cells, and their transformation into trophozoites. Am J Trop Med Hyg. 1983 Jul;32(4):685–690. doi: 10.4269/ajtmh.1983.32.685. [DOI] [PubMed] [Google Scholar]
  9. Hyman B. C., Macinnis A. J. Rapid detection of malaria and other bloodstream parasites by fluorescence microscopy with 4'6 diamidino-2-phenylindole (DAPI). J Parasitol. 1979 Jun;65(3):421–425. [PubMed] [Google Scholar]
  10. Khan Z. M., Vanderberg J. P. Eosinophil-rich, granulomatous inflammatory response to Plasmodium berghei hepatic schizonts in nonimmunized rats is age-related. Am J Trop Med Hyg. 1991 Aug;45(2):190–201. doi: 10.4269/ajtmh.1991.45.190. [DOI] [PubMed] [Google Scholar]
  11. Krotoski W. A., Collins W. E., Jumper J. R. Detection of early exoerythrocytic schizonts of Plasmodium cynomolgi by immunofluorescence. Am J Trop Med Hyg. 1973 Jul;22(4):443–451. doi: 10.4269/ajtmh.1973.22.443. [DOI] [PubMed] [Google Scholar]
  12. Kumar R. K. Immunogold-silver cytochemistry using a capillary action staining system. J Histochem Cytochem. 1989 Jun;37(6):913–917. doi: 10.1177/37.6.2470811. [DOI] [PubMed] [Google Scholar]
  13. Meis J. F., Verhave J. P., Jap P. H., Meuwissen J. H. An ultrastructural study on the role of Kupffer cells in the process of infection by Plasmodium berghei sporozoites in rats. Parasitology. 1983 Apr;86(Pt 2):231–242. doi: 10.1017/s003118200005040x. [DOI] [PubMed] [Google Scholar]
  14. Meis J. F., Verhave J. P., Jap P. H., Meuwissen J. H. Transformation of sporozoites of Plasmodium berghei into exoerythrocytic forms in the liver of its mammalian host. Cell Tissue Res. 1985;241(2):353–360. doi: 10.1007/BF00217180. [DOI] [PubMed] [Google Scholar]
  15. Meis J. F., Verhave J. P., Jap P. H., Sinden R. E., Meuwissen J. H. Malaria parasites--discovery of the early liver form. 1983 Mar 31-Apr 6Nature. 302(5907):424–426. doi: 10.1038/302424a0. [DOI] [PubMed] [Google Scholar]
  16. Pollard K., Lunny D., Holgate C. S., Jackson P., Bird C. C. Fixation, processing, and immunochemical reagent effects on preservation of T-lymphocyte surface membrane antigens in paraffin-embedded tissue. J Histochem Cytochem. 1987 Nov;35(11):1329–1338. doi: 10.1177/35.11.3309048. [DOI] [PubMed] [Google Scholar]
  17. Romero P., Maryanski J. L., Corradin G., Nussenzweig R. S., Nussenzweig V., Zavala F. Cloned cytotoxic T cells recognize an epitope in the circumsporozoite protein and protect against malaria. Nature. 1989 Sep 28;341(6240):323–326. doi: 10.1038/341323a0. [DOI] [PubMed] [Google Scholar]
  18. Smith J. E., Meis J. F., Ponnudurai T., Verhave J. P., Moshage H. J. In-vitro culture of exoerythrocytic form of Plasmodium falciparum in adult human hepatocytes. Lancet. 1984 Sep 29;2(8405):757–758. doi: 10.1016/s0140-6736(84)92670-9. [DOI] [PubMed] [Google Scholar]
  19. Stewart M. J., Vanderberg J. P. Malaria sporozoites release circumsporozoite protein from their apical end and translocate it along their surface. J Protozool. 1991 Jul-Aug;38(4):411–421. doi: 10.1111/j.1550-7408.1991.tb01379.x. [DOI] [PubMed] [Google Scholar]
  20. Suhrbier A., Hamilton A. J., Nicholas J., Sinden R. E. The fate of the circumsporozoite antigens during the exoerythrocytic stage of Plasmodium berghei. Eur J Cell Biol. 1988 Apr;46(1):25–30. [PubMed] [Google Scholar]
  21. Vanderberg J. P., Chew S., Stewart M. J. Plasmodium sporozoite interactions with macrophages in vitro: a videomicroscopic analysis. J Protozool. 1990 Nov-Dec;37(6):528–536. doi: 10.1111/j.1550-7408.1990.tb01260.x. [DOI] [PubMed] [Google Scholar]
  22. Vanderberg J. P. Plasmodium berghei: quantitation of sporozoites injected by mosquitoes feeding on a rodent host. Exp Parasitol. 1977 Jun;42(1):169–181. doi: 10.1016/0014-4894(77)90075-3. [DOI] [PubMed] [Google Scholar]
  23. Vanderberg J., Nussenzweig R., Most H. Protective immunity produced by the injection of x-irradiated sporozoites of Plasmodium berghei. V. In vitro effects of immune serum on sporozoites. Mil Med. 1969 Sep;134(10):1183–1190. [PubMed] [Google Scholar]
  24. Weiss M. M., Vanderberg J. P. Studies on Plasmodium ookinetes. 1. Isolation and concentration from mosquito midguts. J Protozool. 1976 Nov;23(4):547–551. doi: 10.1111/j.1550-7408.1976.tb03837.x. [DOI] [PubMed] [Google Scholar]
  25. Weiss W. R., Mellouk S., Houghten R. A., Sedegah M., Kumar S., Good M. F., Berzofsky J. A., Miller L. H., Hoffman S. L. Cytotoxic T cells recognize a peptide from the circumsporozoite protein on malaria-infected hepatocytes. J Exp Med. 1990 Mar 1;171(3):763–773. doi: 10.1084/jem.171.3.763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yoshida N., Nussenzweig R. S., Potocnjak P., Nussenzweig V., Aikawa M. Hybridoma produces protective antibodies directed against the sporozoite stage of malaria parasite. Science. 1980 Jan 4;207(4426):71–73. doi: 10.1126/science.6985745. [DOI] [PubMed] [Google Scholar]

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