Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1992 Oct 1;89(19):9176–9180. doi: 10.1073/pnas.89.19.9176

Characterization of Plasmodium falciparum sporozoite surface protein 2.

W O Rogers 1, A Malik 1, S Mellouk 1, K Nakamura 1, M D Rogers 1, A Szarfman 1, D M Gordon 1, A K Nussler 1, M Aikawa 1, S L Hoffman 1
PMCID: PMC50088  PMID: 1409621

Abstract

Immunization of mice with Plasmodium yoelii sporozoite surface protein 2 (PySSP2) and circumsporozoite protein protects completely against P. yoelii. The amino acid sequence of PySSP2 suggested that the thrombospondin-related anonymous protein (TRAP) [Robson, K. J. H., Hall, J. R. S., Jennings, M. W., Harris, T. J. R., Marsh, K., Newbold, C. I., Tate, V. E. & Weatherall, D. J. (1988) Nature (London) 335, 79-82] is the Plasmodium falciparum homolog of PySSP2. We report data confirming that TRAP is P. falciparum SSP2 (PfSSP2). Murine antibodies against recombinant PfSSP2 identify a 90-kDa protein in extracts of P. falciparum sporozoites, recognize sporozoites and infected hepatocytes by immunofluorescence, localize PfSSP2 to the sporozoite micronemes by immunoelectron microscopy and to the surface membrane by live immunofluorescence, and inhibit sporozoite invasion and development in hepatocytes in vitro. Human volunteers immunized with irradiated sporozoites and protected against malaria develop antibody and proliferative T-cell responses to PfSSP2, suggesting that, like PySSP2, PfSSP2 is a target of protective immunity, and supporting inclusion of PfSSP2 in a multicomponent malaria vaccine.

Full text

PDF
9176

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abou-Zeid C., Filley E., Steele J., Rook G. A. A simple new method for using antigens separated by polyacrylamide gel electrophoresis to stimulate lymphocytes in vitro after converting bands cut from Western blots into antigen-bearing particles. J Immunol Methods. 1987 Apr 2;98(1):5–10. doi: 10.1016/0022-1759(87)90429-7. [DOI] [PubMed] [Google Scholar]
  2. Aikawa M., Atkinson C. T., Beaudoin L. M., Sedegah M., Charoenvit Y., Beaudoin R. Localization of CS and non-CS antigens in the sporogonic stages of Plasmodium yoelii. Bull World Health Organ. 1990;68 (Suppl):165–171. [PMC free article] [PubMed] [Google Scholar]
  3. Charoenvit Y., Leef M. F., Yuan L. F., Sedegah M., Beaudoin R. L. Characterization of Plasmodium yoelii monoclonal antibodies directed against stage-specific sporozoite antigens. Infect Immun. 1987 Mar;55(3):604–608. doi: 10.1128/iai.55.3.604-608.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Clarke L. E., Tomley F. M., Wisher M. H., Foulds I. J., Boursnell M. E. Regions of an Eimeria tenella antigen contain sequences which are conserved in circumsporozoite proteins from Plasmodium spp. and which are related to the thrombospondin gene family. Mol Biochem Parasitol. 1990 Jun;41(2):269–279. doi: 10.1016/0166-6851(90)90190-w. [DOI] [PubMed] [Google Scholar]
  6. Clyde D. F. Immunization of man against falciparum and vivax malaria by use of attenuated sporozoites. Am J Trop Med Hyg. 1975 May;24(3):397–401. doi: 10.4269/ajtmh.1975.24.397. [DOI] [PubMed] [Google Scholar]
  7. Dame J. B., Williams J. L., McCutchan T. F., Weber J. L., Wirtz R. A., Hockmeyer W. T., Maloy W. L., Haynes J. D., Schneider I., Roberts D. Structure of the gene encoding the immunodominant surface antigen on the sporozoite of the human malaria parasite Plasmodium falciparum. Science. 1984 Aug 10;225(4662):593–599. doi: 10.1126/science.6204383. [DOI] [PubMed] [Google Scholar]
  8. Hedstrom R. C., Campbell J. R., Leef M. L., Charoenvit Y., Carter M., Sedegah M., Beaudoin R. L., Hoffman S. L. A malaria sporozoite surface antigen distinct from the circumsporozoite protein. Bull World Health Organ. 1990;68 (Suppl):152–157. [PMC free article] [PubMed] [Google Scholar]
  9. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  10. Herrington D. A., Nardin E. H., Losonsky G., Bathurst I. C., Barr P. J., Hollingdale M. R., Edelman R., Levine M. M. Safety and immunogenicity of a recombinant sporozoite malaria vaccine against Plasmodium vivax. Am J Trop Med Hyg. 1991 Dec;45(6):695–701. doi: 10.4269/ajtmh.1991.45.695. [DOI] [PubMed] [Google Scholar]
  11. Khusmith S., Charoenvit Y., Kumar S., Sedegah M., Beaudoin R. L., Hoffman S. L. Protection against malaria by vaccination with sporozoite surface protein 2 plus CS protein. Science. 1991 May 3;252(5006):715–718. doi: 10.1126/science.1827210. [DOI] [PubMed] [Google Scholar]
  12. Kumar S., Miller L. H., Quakyi I. A., Keister D. B., Houghten R. A., Maloy W. L., Moss B., Berzofsky J. A., Good M. F. Cytotoxic T cells specific for the circumsporozoite protein of Plasmodium falciparum. Nature. 1988 Jul 21;334(6179):258–260. doi: 10.1038/334258a0. [DOI] [PubMed] [Google Scholar]
  13. Lal A. A., de la Cruz V. F., Welsh J. A., Charoenvit Y., Maloy W. L., McCutchan T. F. Structure of the gene encoding the circumsporozoite protein of Plasmodium yoelii. A rodent model for examining antimalarial sporozoite vaccines. J Biol Chem. 1987 Mar 5;262(7):2937–2940. [PubMed] [Google Scholar]
  14. Malik A., Egan J. E., Houghten R. A., Sadoff J. C., Hoffman S. L. Human cytotoxic T lymphocytes against the Plasmodium falciparum circumsporozoite protein. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3300–3304. doi: 10.1073/pnas.88.8.3300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mellouk S., Berbiguier N., Druilhe P., Sedegah M., Galey B., Yuan L., Leef M., Charoenvit Y., Paul C., Hoffman S. Evaluation of an in vitro assay aimed at measuring protective antibodies against sporozoites. Bull World Health Organ. 1990;68 (Suppl):52–59. [PMC free article] [PubMed] [Google Scholar]
  16. Mellouk S., Maheshwari R. K., Rhodes-Feuillette A., Beaudoin R. L., Berbiguier N., Matile H., Miltgen F., Landau I., Pied S., Chigot J. P. Inhibitory activity of interferons and interleukin 1 on the development of Plasmodium falciparum in human hepatocyte cultures. J Immunol. 1987 Dec 15;139(12):4192–4195. [PubMed] [Google Scholar]
  17. Nussenzweig R. S., Vanderberg J., Most H., Orton C. Protective immunity produced by the injection of x-irradiated sporozoites of plasmodium berghei. Nature. 1967 Oct 14;216(5111):160–162. doi: 10.1038/216160a0. [DOI] [PubMed] [Google Scholar]
  18. Potocnjak P., Yoshida N., Nussenzweig R. S., Nussenzweig V. Monovalent fragments (Fab) of monoclonal antibodies to a sporozoite surface antigen (Pb44) protect mice against malarial infection. J Exp Med. 1980 Jun 1;151(6):1504–1513. doi: 10.1084/jem.151.6.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rieckmann K. H., Carson P. E., Beaudoin R. L., Cassells J. S., Sell K. W. Letter: Sporozoite induced immunity in man against an Ethiopian strain of Plasmodium falciparum. Trans R Soc Trop Med Hyg. 1974;68(3):258–259. doi: 10.1016/0035-9203(74)90129-1. [DOI] [PubMed] [Google Scholar]
  20. Robson K. J., Hall J. R., Davies L. C., Crisanti A., Hill A. V., Wellems T. E. Polymorphism of the TRAP gene of Plasmodium falciparum. Proc Biol Sci. 1990 Dec 22;242(1305):205–216. doi: 10.1098/rspb.1990.0126. [DOI] [PubMed] [Google Scholar]
  21. Robson K. J., Hall J. R., Jennings M. W., Harris T. J., Marsh K., Newbold C. I., Tate V. E., Weatherall D. J. A highly conserved amino-acid sequence in thrombospondin, properdin and in proteins from sporozoites and blood stages of a human malaria parasite. Nature. 1988 Sep 1;335(6185):79–82. doi: 10.1038/335079a0. [DOI] [PubMed] [Google Scholar]
  22. Rogers W. O., Rogers M. D., Hedstrom R. C., Hoffman S. L. Characterization of the gene encoding sporozoite surface protein 2, a protective Plasmodium yoelii sporozoite antigen. Mol Biochem Parasitol. 1992 Jul;53(1-2):45–51. doi: 10.1016/0166-6851(92)90005-5. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Rénia L., Miltgen F., Charoenvit Y., Ponnudurai T., Verhave J. P., Collins W. E., Mazier D. Malaria sporozoite penetration. A new approach by double staining. J Immunol Methods. 1988 Sep 13;112(2):201–205. doi: 10.1016/0022-1759(88)90358-4. [DOI] [PubMed] [Google Scholar]
  25. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  27. Szarfman A., Walliker D., McBride J. S., Lyon J. A., Quakyi I. A., Carter R. Allelic forms of gp195, a major blood-stage antigen of Plasmodium falciparum, are expressed in liver stages. J Exp Med. 1988 Jan 1;167(1):231–236. doi: 10.1084/jem.167.1.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tomley F. M., Clarke L. E., Kawazoe U., Dijkema R., Kok J. J. Sequence of the gene encoding an immunodominant microneme protein of Eimeria tenella. Mol Biochem Parasitol. 1991 Dec;49(2):277–288. doi: 10.1016/0166-6851(91)90071-d. [DOI] [PubMed] [Google Scholar]
  29. Walliker D., Quakyi I. A., Wellems T. E., McCutchan T. F., Szarfman A., London W. T., Corcoran L. M., Burkot T. R., Carter R. Genetic analysis of the human malaria parasite Plasmodium falciparum. Science. 1987 Jun 26;236(4809):1661–1666. doi: 10.1126/science.3299700. [DOI] [PubMed] [Google Scholar]
  30. 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]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES