Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1996 Jul;64(7):2602–2608. doi: 10.1128/iai.64.7.2602-2608.1996

Influence of adjuvants on protection induced by a recombinant fusion protein against malarial infection.

T M Daly 1, C A Long 1
PMCID: PMC174116  PMID: 8698485

Abstract

Previously, we described a protective immune response induced by the carboxyl-terminal region of the merozoite surface protein-1 (MSP-1) from the rodent malarial parasite Plasmodium yoelii yoelii 17XL, expressed as a fusion protein and designated glutathione S-transferase (GST)-PYC2. We also demonstrated that the humoral response induced by GST-PYC2 was the primary mechanism by which immunized animals controlled their blood-stage infections. We have now examined the influence of several adjuvants on the immune response to the GST-PYC2 fusion protein. While alum, Freund's adjuvant, Ribi adjuvant system, and TiterMax were efficacious in eliciting a protective response with GST-PYC2 in BALB/c mice, saponin failed to induce protection, although significant levels of PYC2-specific antibodies were produced in all immunized animals. This protection depended on the mouse strain since immunization of Swiss Webster mice with GST-PYC2 in alum did not produce levels of PYC2-specific antibodies comparable to those in BALB/c mice nor did it induce any demonstrable level of protection against parasite challenge. Swiss Webster mice were protected, however, when immunized with GST-PYC2 in other adjuvants. Immunization with PYC2, isolated free of GST induced lower levels of antigen-specific antibody; only those animals given PYC2 in Freund's adjuvant demonstrated a significant degree of protection, suggesting the possibility (of additional cellular effector mechanisms. These findings demonstrate that adjuvant, host genotype, and the fine specificity of the response significantly influence the protection induced by the carboxyl terminus of MSP-1 in vivo and illustrate the need to consider these factors in evaluating MSP-1 as a vaccine component.

Full Text

The Full Text of this article is available as a PDF (294.4 KB).

Selected References

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

  1. Allison A. C., Byars N. E. An adjuvant formulation that selectively elicits the formation of antibodies of protective isotypes and of cell-mediated immunity. J Immunol Methods. 1986 Dec 24;95(2):157–168. doi: 10.1016/0022-1759(86)90402-3. [DOI] [PubMed] [Google Scholar]
  2. Blackman M. J., Heidrich H. G., Donachie S., McBride J. S., Holder A. A. A single fragment of a malaria merozoite surface protein remains on the parasite during red cell invasion and is the target of invasion-inhibiting antibodies. J Exp Med. 1990 Jul 1;172(1):379–382. doi: 10.1084/jem.172.1.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blackman M. J., Scott-Finnigan T. J., Shai S., Holder A. A. Antibodies inhibit the protease-mediated processing of a malaria merozoite surface protein. J Exp Med. 1994 Jul 1;180(1):389–393. doi: 10.1084/jem.180.1.389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burns J. M., Jr, Daly T. M., Vaidya A. B., Long C. A. The 3' portion of the gene for a Plasmodium yoelii merozoite surface antigen encodes the epitope recognized by a protective monoclonal antibody. Proc Natl Acad Sci U S A. 1988 Jan;85(2):602–606. doi: 10.1073/pnas.85.2.602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burns J. M., Jr, Majarian W. R., Young J. F., Daly T. M., Long C. A. A protective monoclonal antibody recognizes an epitope in the carboxyl-terminal cysteine-rich domain in the precursor of the major merozoite surface antigen of the rodent malarial parasite, Plasmodium yoelii. J Immunol. 1989 Oct 15;143(8):2670–2676. [PubMed] [Google Scholar]
  6. Chang S. P., Gibson H. L., Lee-Ng C. T., Barr P. J., Hui G. S. A carboxyl-terminal fragment of Plasmodium falciparum gp195 expressed by a recombinant baculovirus induces antibodies that completely inhibit parasite growth. J Immunol. 1992 Jul 15;149(2):548–555. [PubMed] [Google Scholar]
  7. Chang S. P., Nikaido C. M., Hashimoto A. C., Hashiro C. Q., Yokota B. T., Hui G. S. Regulation of antibody specificity to Plasmodium falciparum merozoite surface protein-1 by adjuvant and MHC haplotype. J Immunol. 1994 Apr 1;152(7):3483–3490. [PubMed] [Google Scholar]
  8. Chappel J. A., Holder A. A. Monoclonal antibodies that inhibit Plasmodium falciparum invasion in vitro recognise the first growth factor-like domain of merozoite surface protein-1. Mol Biochem Parasitol. 1993 Aug;60(2):303–311. doi: 10.1016/0166-6851(93)90141-j. [DOI] [PubMed] [Google Scholar]
  9. Daly T. M., Burns J. M., Jr, Long C. A. Comparison of the carboxy-terminal, cysteine-rich domain of the merozoite surface protein-1 from several strains of Plasmodium yoelii. Mol Biochem Parasitol. 1992 Jun;52(2):279–282. doi: 10.1016/0166-6851(92)90061-n. [DOI] [PubMed] [Google Scholar]
  10. Daly T. M., Long C. A. A recombinant 15-kilodalton carboxyl-terminal fragment of Plasmodium yoelii yoelii 17XL merozoite surface protein 1 induces a protective immune response in mice. Infect Immun. 1993 Jun;61(6):2462–2467. doi: 10.1128/iai.61.6.2462-2467.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Daly T. M., Long C. A. Humoral response to a carboxyl-terminal region of the merozoite surface protein-1 plays a predominant role in controlling blood-stage infection in rodent malaria. J Immunol. 1995 Jul 1;155(1):236–243. [PubMed] [Google Scholar]
  12. Etlinger H. M., Caspers P., Matile H., Schoenfeld H. J., Stueber D., Takacs B. Ability of recombinant or native proteins to protect monkeys against heterologous challenge with Plasmodium falciparum. Infect Immun. 1991 Oct;59(10):3498–3503. doi: 10.1128/iai.59.10.3498-3503.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Farley P. J., Long C. A. Plasmodium yoelii yoelii 17XL MSP-1: fine-specificity mapping of a discontinuous, disulfide-dependent epitope recognized by a protective monoclonal antibody using expression PCR (E-PCR). Exp Parasitol. 1995 Mar;80(2):328–332. doi: 10.1006/expr.1995.1040. [DOI] [PubMed] [Google Scholar]
  14. Freeman R. R., Holder A. A. Characteristics of the protective response of BALB/c mice immunized with a purified Plasmodium yoelii schizont antigen. Clin Exp Immunol. 1983 Dec;54(3):609–616. [PMC free article] [PubMed] [Google Scholar]
  15. Grun J. L., Maurer P. H. Different T helper cell subsets elicited in mice utilizing two different adjuvant vehicles: the role of endogenous interleukin 1 in proliferative responses. Cell Immunol. 1989 Jun;121(1):134–145. doi: 10.1016/0008-8749(89)90011-7. [DOI] [PubMed] [Google Scholar]
  16. Holder A. A., Freeman R. R. Immunization against blood-stage rodent malaria using purified parasite antigens. Nature. 1981 Nov 26;294(5839):361–364. doi: 10.1038/294361a0. [DOI] [PubMed] [Google Scholar]
  17. Hui G. S., Chang S. P., Gibson H., Hashimoto A., Hashiro C., Barr P. J., Kotani S. Influence of adjuvants on the antibody specificity to the Plasmodium falciparum major merozoite surface protein, gp195. J Immunol. 1991 Dec 1;147(11):3935–3941. [PubMed] [Google Scholar]
  18. Hui G. S., Gosnell W. L., Case S. E., Hashiro C., Nikaido C., Hashimoto A., Kaslow D. C. Immunogenicity of the C-terminal 19-kDa fragment of the Plasmodium falciparum merozoite surface protein 1 (MSP1), YMSP1(19) expressed in S. cerevisiae. J Immunol. 1994 Sep 15;153(6):2544–2553. [PubMed] [Google Scholar]
  19. Hui G. S., Hashimoto A., Chang S. P. Roles of conserved and allelic regions of the major merozoite surface protein (gp195) in immunity against Plasmodium falciparum. Infect Immun. 1992 Apr;60(4):1422–1433. doi: 10.1128/iai.60.4.1422-1433.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hui G. S., Hashiro C., Nikaido C., Case S. E., Hashimoto A., Gibson H., Barr P. J., Chang S. P. Immunological cross-reactivity of the C-terminal 42-kilodalton fragment of Plasmodium falciparum merozoite surface protein 1 expressed in baculovirus. Infect Immun. 1993 Aug;61(8):3403–3411. doi: 10.1128/iai.61.8.3403-3411.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hui G. S., Tam L. Q., Chang S. P., Case S. E., Hashiro C., Siddiqui W. A., Shiba T., Kusumoto S., Kotani S. Synthetic low-toxicity muramyl dipeptide and monophosphoryl lipid A replace Freund complete adjuvant in inducing growth-inhibitory antibodies to the Plasmodium falciparum major merozoite surface protein, gp195. Infect Immun. 1991 May;59(5):1585–1591. doi: 10.1128/iai.59.5.1585-1591.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hunter R. L., Lal A. A. Copolymer adjuvants in malaria vaccine development. Am J Trop Med Hyg. 1994;50(4 Suppl):52–58. doi: 10.4269/ajtmh.1994.50.52. [DOI] [PubMed] [Google Scholar]
  23. Jongwutiwes S., Tanabe K., Kanbara H. Sequence conservation in the C-terminal part of the precursor to the major merozoite surface proteins (MSP1) of Plasmodium falciparum from field isolates. Mol Biochem Parasitol. 1993 May;59(1):95–100. doi: 10.1016/0166-6851(93)90010-u. [DOI] [PubMed] [Google Scholar]
  24. Kang Y., Long C. A. Sequence heterogeneity of the C-terminal, Cys-rich region of the merozoite surface protein-1 (MSP-1) in field samples of Plasmodium falciparum. Mol Biochem Parasitol. 1995 Jul;73(1-2):103–110. doi: 10.1016/0166-6851(95)00102-7. [DOI] [PubMed] [Google Scholar]
  25. Karagouni E. E., Hadjipetrou-Kourounakis L. Regulation of isotype immunoglobulin production by adjuvants in vivo. Scand J Immunol. 1990 Jun;31(6):745–754. doi: 10.1111/j.1365-3083.1990.tb02826.x. [DOI] [PubMed] [Google Scholar]
  26. Kenney J. S., Hughes B. W., Masada M. P., Allison A. C. Influence of adjuvants on the quantity, affinity, isotype and epitope specificity of murine antibodies. J Immunol Methods. 1989 Jul 26;121(2):157–166. doi: 10.1016/0022-1759(89)90156-7. [DOI] [PubMed] [Google Scholar]
  27. Kumar S., Yadava A., Keister D. B., Tian J. H., Ohl M., Perdue-Greenfield K. A., Miller L. H., Kaslow D. C. Immunogenicity and in vivo efficacy of recombinant Plasmodium falciparum merozoite surface protein-1 in Aotus monkeys. Mol Med. 1995 Mar;1(3):325–332. [PMC free article] [PubMed] [Google Scholar]
  28. Lewis A. P. Cloning and analysis of the gene encoding the 230-kilodalton merozoite surface antigen of Plasmodium yoelii. Mol Biochem Parasitol. 1989 Oct;36(3):271–282. doi: 10.1016/0166-6851(89)90175-8. [DOI] [PubMed] [Google Scholar]
  29. Ling I. T., Ogun S. A., Holder A. A. Immunization against malaria with a recombinant protein. Parasite Immunol. 1994 Feb;16(2):63–67. doi: 10.1111/j.1365-3024.1994.tb00324.x. [DOI] [PubMed] [Google Scholar]
  30. Long C. A. Immunity to blood stages of malaria. Curr Opin Immunol. 1993 Aug;5(4):548–556. doi: 10.1016/0952-7915(93)90036-r. [DOI] [PubMed] [Google Scholar]
  31. Majarian W. R., Daly T. M., Weidanz W. P., Long C. A. Passive immunization against murine malaria with an IgG3 monoclonal antibody. J Immunol. 1984 Jun;132(6):3131–3137. [PubMed] [Google Scholar]
  32. Miller L. H., Roberts T., Shahabuddin M., McCutchan T. F. Analysis of sequence diversity in the Plasmodium falciparum merozoite surface protein-1 (MSP-1). Mol Biochem Parasitol. 1993 May;59(1):1–14. doi: 10.1016/0166-6851(93)90002-f. [DOI] [PubMed] [Google Scholar]
  33. Nussenzweig R. S., Long C. A. Malaria vaccines: multiple targets. Science. 1994 Sep 2;265(5177):1381–1383. doi: 10.1126/science.8073276. [DOI] [PubMed] [Google Scholar]
  34. Siddiqui W. A., Tam L. Q., Kramer K. J., Hui G. S., Case S. E., Yamaga K. M., Chang S. P., Chan E. B., Kan S. C. Merozoite surface coat precursor protein completely protects Aotus monkeys against Plasmodium falciparum malaria. Proc Natl Acad Sci U S A. 1987 May;84(9):3014–3018. doi: 10.1073/pnas.84.9.3014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Varley C. A., Dunne D. W., Havercroft J. C. The influence of adjuvant on humoral responses to glutathione-S-transferase fusion proteins. Parasite Immunol. 1992 Sep;14(5):557–562. doi: 10.1111/j.1365-3024.1992.tb00028.x. [DOI] [PubMed] [Google Scholar]
  37. White W. I., Evans C. B., Taylor D. W. Antimalarial antibodies of the immunoglobulin G2a isotype modulate parasitemias in mice infected with Plasmodium yoelii. Infect Immun. 1991 Oct;59(10):3547–3554. doi: 10.1128/iai.59.10.3547-3554.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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