Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1995 Feb;63(2):554–562. doi: 10.1128/iai.63.2.554-562.1995

Induction of opsonizing antibodies after injection of recombinant Plasmodium falciparum vaccine candidate antigens in preimmune Saimiri sciureus monkeys.

R Perraut 1, O Mercereau-Puijalon 1, D Mattei 1, E Bourreau 1, O Garraud 1, B Bonnemains 1, L Pereia de Silva 1, J C Michel 1
PMCID: PMC173031  PMID: 7822021

Abstract

We have previously shown that Plasmodium falciparum recombinant antigens PfEB200, R23, and Pfi72 inhibit opsonization of infected erythrocytes by hyperimmune Saimiri sera, indicating that they contain target epitopes involved in the phagocytosis of infected erythrocytes. We have investigated in this study the immune response of Saimiri monkeys with previous experience of malaria infections (preimmune monkeys) after injection of these recombinant antigens, administered alone or simultaneously. The humoral response to the recombinant antigens was monitored by radioimmunoassay, and the response to P. falciparum blood stages was assayed by immunofluorescence. The relative proportion of protective versus nonprotective immunoglobulin subtypes was investigated by using 3A2/G6 and 3E4/H8 monoclonal antibodies, and the capacity of the antisera to promote in vitro phagocytosis of infected erythrocytes was evaluated. The antigens evoked in most cases a secondary-type antibody response, resulting in important increases in antigen-specific antibody titers and concomitantly in anti-P. falciparum titers. The ratio of 3A2/G6 to 3E4/H8 immunoglobulin subtypes varied with the immunogen used. Opsonizing antibodies were boosted in several animals, the most promising combination being the mixture of PfEB200 and R23 that induced long-lasting production in five of five animals. The detectable opsonizing activity appearing after immunization of the animals was antigen specific, as it was lost after adsorption of the recombinant antigens. The challenge of the animals with blood stage parasites confirmed previous findings showing a correlation between the presence of detectable opsonizing antibodies in serum and protection.

Full Text

The Full Text of this article is available as a PDF (250.2 KB).

Selected References

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

  1. Barrios C., Lussow A. R., Van Embden J., Van der Zee R., Rappuoli R., Costantino P., Louis J. A., Lambert P. H., Del Giudice G. Mycobacterial heat-shock proteins as carrier molecules. II: The use of the 70-kDa mycobacterial heat-shock protein as carrier for conjugated vaccines can circumvent the need for adjuvants and Bacillus Calmette Guérin priming. Eur J Immunol. 1992 Jun;22(6):1365–1372. doi: 10.1002/eji.1830220606. [DOI] [PubMed] [Google Scholar]
  2. Bonnefoy S., Guillotte M., Langsley G., Mercereau-Puijalon O. Plasmodium falciparum: characterization of gene R45 encoding a trophozoite antigen containing a central block of six amino acid repeats. Exp Parasitol. 1992 Jun;74(4):441–451. doi: 10.1016/0014-4894(92)90206-p. [DOI] [PubMed] [Google Scholar]
  3. Bouharoun-Tayoun H., Attanath P., Sabchareon A., Chongsuphajaisiddhi T., Druilhe P. Antibodies that protect humans against Plasmodium falciparum blood stages do not on their own inhibit parasite growth and invasion in vitro, but act in cooperation with monocytes. J Exp Med. 1990 Dec 1;172(6):1633–1641. doi: 10.1084/jem.172.6.1633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bouharoun-Tayoun H., Druilhe P. Plasmodium falciparum malaria: evidence for an isotype imbalance which may be responsible for delayed acquisition of protective immunity. Infect Immun. 1992 Apr;60(4):1473–1481. doi: 10.1128/iai.60.4.1473-1481.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown J., Greenwood B. M. Cellular and humoral inhibition of Plasmodium falciparum growth in vitro and recovery from acute malaria. Parasite Immunol. 1985 May;7(3):265–275. doi: 10.1111/j.1365-3024.1985.tb00075.x. [DOI] [PubMed] [Google Scholar]
  6. Druilhe P., Khusmith S. Epidemiological correlation between levels of antibodies promoting merozoite phagocytosis of Plasmodium falciparum and malaria-immune status. Infect Immun. 1987 Apr;55(4):888–891. doi: 10.1128/iai.55.4.888-891.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fandeur T., Dubois P., Gysin J., Dedet J. P., da Silva L. P. In vitro and in vivo studies on protective and inhibitory antibodies against Plasmodium falciparum in the Saimiri monkey. J Immunol. 1984 Jan;132(1):432–437. [PubMed] [Google Scholar]
  8. Groux H., Gysin J. Opsonization as an effector mechanism in human protection against asexual blood stages of Plasmodium falciparum: functional role of IgG subclasses. Res Immunol. 1990 Jul-Aug;141(6):529–542. doi: 10.1016/0923-2494(90)90021-p. [DOI] [PubMed] [Google Scholar]
  9. Groux H., Perraut R., Garraud O., Poingt J. P., Gysin J. Functional characterization of the antibody-mediated protection against blood stages of Plasmodium falciparum in the monkey Saimiri sciureus. Eur J Immunol. 1990 Oct;20(10):2317–2323. doi: 10.1002/eji.1830201022. [DOI] [PubMed] [Google Scholar]
  10. Guiguemdé T. R., Sturchler D., Ouédraogo J. B., Drabo M., Etlinger H., Douchet C., Gbary A. R., Haller L., Kambou S., Fernex M. Vaccination contre le paludisme: premier essai avec un vaccin antisporozoïte, le (NANP)3-TT (RO 40-2361) en Afrique (Bobo-Dioulasso, Burkina Faso). Bull Soc Pathol Exot. 1990;83(2):217–227. [PubMed] [Google Scholar]
  11. Gysin J., Gavoille S., Mattei D., Scherf A., Bonnefoy S., Mercereau-Puijalon O., Feldmann T., Kun J., Müller-Hill B., Pereira da Silva L. In vitro phagocytosis inhibition assay for the screening of potential candidate antigens for sub-unit vaccines against the asexual blood stage of Plasmodium falciparum. J Immunol Methods. 1993 Feb 26;159(1-2):209–219. doi: 10.1016/0022-1759(93)90159-5. [DOI] [PubMed] [Google Scholar]
  12. Gysin J., Hommel M., da Silva L. P. Experimental infection of the squirrel monkey (Saimiri sciureus) with Plasmodium falciparum. J Parasitol. 1980 Dec;66(6):1003–1009. [PubMed] [Google Scholar]
  13. Gysin J., Pauillac S., Fandeur T. Characterization by anti-Ig monoclonal antibodies of protective and non-protective antibodies against asexual forms of Plasmodium falciparum in the Saimiri monkey. Ann Inst Pasteur Immunol. 1987 Nov-Dec;138(6):829–844. doi: 10.1016/s0769-2625(87)80003-x. [DOI] [PubMed] [Google Scholar]
  14. Hinterberg K., Scherf A., Gysin J., Toyoshima T., Aikawa M., Mazie J. C., da Silva L. P., Mattei D. Plasmodium falciparum: the Pf332 antigen is secreted from the parasite by a brefeldin A-dependent pathway and is translocated to the erythrocyte membrane via the Maurer's clefts. Exp Parasitol. 1994 Nov;79(3):279–291. doi: 10.1006/expr.1994.1091. [DOI] [PubMed] [Google Scholar]
  15. Hughes A. L. Conserved proteins as immunogens: glutathione S-transferase of Schistosoma. Parasitol Today. 1994 Apr;10(4):149–151. doi: 10.1016/0169-4758(94)90266-6. [DOI] [PubMed] [Google Scholar]
  16. Knapp B., Hundt E., Enders B., Küpper H. A. Protection of Aotus monkeys from malaria infection by immunization with recombinant hybrid proteins. Infect Immun. 1992 Jun;60(6):2397–2401. doi: 10.1128/iai.60.6.2397-2401.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lunel F., Druilhe P. Effector cells involved in nonspecific and antibody-dependent mechanisms directed against Plasmodium falciparum blood stages in vitro. Infect Immun. 1989 Jul;57(7):2043–2049. doi: 10.1128/iai.57.7.2043-2049.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marsh K., Otoo L., Hayes R. J., Carson D. C., Greenwood B. M. Antibodies to blood stage antigens of Plasmodium falciparum in rural Gambians and their relation to protection against infection. Trans R Soc Trop Med Hyg. 1989 May-Jun;83(3):293–303. doi: 10.1016/0035-9203(89)90478-1. [DOI] [PubMed] [Google Scholar]
  19. Mattei D., Scherf A., Bensaude O., da Silva L. P. A heat shock-like protein from the human malaria parasite Plasmodium falciparum induces autoantibodies. Eur J Immunol. 1989 Oct;19(10):1823–1828. doi: 10.1002/eji.1830191010. [DOI] [PubMed] [Google Scholar]
  20. Mattei D., Scherf A. The Pf332 gene of Plasmodium falciparum codes for a giant protein that is translocated from the parasite to the membrane of infected erythrocytes. Gene. 1992 Jan 2;110(1):71–79. doi: 10.1016/0378-1119(92)90446-v. [DOI] [PubMed] [Google Scholar]
  21. McGregor I. A. Towards a vaccine against malaria. Br J Biomed Sci. 1993 Mar;50(1):35–42. [PubMed] [Google Scholar]
  22. Michel J. C., Fandeur T., Neuilly G., Roussilhon C., Dedet J. P. Opsonic activity of ascitic fluids from Plasmodium falciparum-infected Saimiri monkey: positive correlation with protection in passive transfer assay. Ann Immunol (Paris) 1983 Nov-Dec;134D(3):373–383. doi: 10.1016/s0769-2625(83)80028-2. [DOI] [PubMed] [Google Scholar]
  23. Perraut R., Chouteau P., Bourreau E., Bonnemains B., Garraud O. Assays for adjuvanticity of new formulations and of carrier proteins for inducing antibody responses to selected immunogens in the squirrel monkey Saimiri sciureus. Immunol Cell Biol. 1994 Apr;72(2):169–175. doi: 10.1038/icb.1994.25. [DOI] [PubMed] [Google Scholar]
  24. Perraut R., Hundt E., Garraud O., Enders B., Gysin J. Comparison of the effects of adjuvants and adjuvant doses on the quantitative and qualitative antibody response to selected antigens in New World squirrel monkeys Saimiri sciureus. Vaccine. 1993;11(7):730–736. doi: 10.1016/0264-410x(93)90257-x. [DOI] [PubMed] [Google Scholar]
  25. Perraut R., Lussow A. R., Gavoille S., Garraud O., Matile H., Tougne C., van Embden J., van der Zee R., Lambert P. H., Gysin J. Successful primate immunization with peptides conjugated to purified protein derivative or mycobacterial heat shock proteins in the absence of adjuvants. Clin Exp Immunol. 1993 Sep;93(3):382–386. doi: 10.1111/j.1365-2249.1993.tb08189.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Roussilhon C., Fandeur T., Dedet J. P. Long-term protection of squirrel monkeys (Saimiri sciureus) against Plasmodium falciparum challenge inoculations after various time intervals. Parasitol Res. 1988;75(2):118–122. doi: 10.1007/BF00932711. [DOI] [PubMed] [Google Scholar]
  27. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  28. Teuscher T., Schellenberg J. R., Bastos de Azevedo I., Hurt N., Smith T., Hayes R., Masanja H., Silva Y., Lopez M. C., Kitua A. SPf66, a chemically synthesized subunit malaria vaccine, is safe and immunogenic in Tanzanians exposed to intense malaria transmission. Vaccine. 1994 Mar;12(4):328–336. doi: 10.1016/0264-410x(94)90097-3. [DOI] [PubMed] [Google Scholar]
  29. Valero M. V., Amador L. R., Galindo C., Figueroa J., Bello M. S., Murillo L. A., Mora A. L., Patarroyo G., Rocha C. L., Rojas M. Vaccination with SPf66, a chemically synthesised vaccine, against Plasmodium falciparum malaria in Colombia. Lancet. 1993 Mar 20;341(8847):705–710. doi: 10.1016/0140-6736(93)90483-w. [DOI] [PubMed] [Google Scholar]
  30. ten Hagen T. L., Sulzer A. J., Kidd M. R., Lal A. A., Hunter R. L. Role of adjuvants in the modulation of antibody isotype, specificity, and induction of protection by whole blood-stage Plasmodium yoelii vaccines. J Immunol. 1993 Dec 15;151(12):7077–7085. [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES