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. 1997 Jul;65(7):2772–2777. doi: 10.1128/iai.65.7.2772-2777.1997

Expression and serologic activity of a soluble recombinant Plasmodium vivax Duffy binding protein.

T Fraser 1, P Michon 1, J W Barnwell 1, A R Noe 1, F Al-Yaman 1, D C Kaslow 1, J H Adams 1
PMCID: PMC175391  PMID: 9199449

Abstract

Plasmodium vivax Duffy binding protein (DBP) is a conserved functionally important protein. P. vivax DBP is an asexual blood-stage malaria vaccine candidate because adhesion of P. vivax DBP to its erythrocyte receptor is essential for the parasite to continue development in human blood. We developed a soluble recombinant protein of P. vivax DBP (rDBP) and examined serologic activity to it in residents of a region of high endemicity. This soluble rDBP product contained the cysteine-rich ligand domain and most of the contiguous proline-rich hydrophilic region. rDBP was expressed as a glutathione S-transferase (GST) fusion protein and was isolated from GST by thrombin treatment of the purified fusion protein bound on glutathione agarose beads. P. vivax rDBP was immunogenic in rabbits and induced antibodies that reacted with P. vivax and Plasmodium knowlesi merozoites. Human sera from adult residents of a region of Papua New Guinea where malaria is highly endemic or P. vivax-infected North American residents reacted with rDBP in an immunoblot and an enzyme-linked immunosorbent assay. The reactivity to reduced, denatured P. vivax rDBP and the cross-reactivity with P. knowlesi indicated the presence of immunogenic conserved linear B-cell epitopes. A more extensive serologic survey of Papua New Guinea residents showed that antibody response to P. vivax DBP is common and increases with age, suggesting a possible boosting of the antibody response in some by repeated exposure to P. vivax. A positive humoral response to P. vivax DBP correlated with a significantly higher response to P. vivax MSP-1(19). The natural immunogenicity of this DBP should strengthen its usefulness as a vaccine.

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Selected References

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  1. Adams J. H., Hudson D. E., Torii M., Ward G. E., Wellems T. E., Aikawa M., Miller L. H. The Duffy receptor family of Plasmodium knowlesi is located within the micronemes of invasive malaria merozoites. Cell. 1990 Oct 5;63(1):141–153. doi: 10.1016/0092-8674(90)90295-p. [DOI] [PubMed] [Google Scholar]
  2. Adams J. H., Sim B. K., Dolan S. A., Fang X., Kaslow D. C., Miller L. H. A family of erythrocyte binding proteins of malaria parasites. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7085–7089. doi: 10.1073/pnas.89.15.7085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ampudia E., Patarroyo M. A., Patarroyo M. E., Murillo L. A. Genetic polymorphism of the Duffy receptor binding domain of Plasmodium vivax in Colombian wild isolates. Mol Biochem Parasitol. 1996 Jun;78(1-2):269–272. doi: 10.1016/s0166-6851(96)02611-4. [DOI] [PubMed] [Google Scholar]
  4. Barnwell J. W., Nichols M. E., Rubinstein P. In vitro evaluation of the role of the Duffy blood group in erythrocyte invasion by Plasmodium vivax. J Exp Med. 1989 May 1;169(5):1795–1802. doi: 10.1084/jem.169.5.1795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baruch D. I., Pasloske B. L., Singh H. B., Bi X., Ma X. C., Feldman M., Taraschi T. F., Howard R. J. Cloning the P. falciparum gene encoding PfEMP1, a malarial variant antigen and adherence receptor on the surface of parasitized human erythrocytes. Cell. 1995 Jul 14;82(1):77–87. doi: 10.1016/0092-8674(95)90054-3. [DOI] [PubMed] [Google Scholar]
  6. Cattani J. A., Tulloch J. L., Vrbova H., Jolley D., Gibson F. D., Moir J. S., Heywood P. F., Alpers M. P., Stevenson A., Clancy R. The epidemiology of malaria in a population surrounding Madang, Papua New Guinea. Am J Trop Med Hyg. 1986 Jan;35(1):3–15. doi: 10.4269/ajtmh.1986.35.3. [DOI] [PubMed] [Google Scholar]
  7. Chitnis C. E., Miller L. H. Identification of the erythrocyte binding domains of Plasmodium vivax and Plasmodium knowlesi proteins involved in erythrocyte invasion. J Exp Med. 1994 Aug 1;180(2):497–506. doi: 10.1084/jem.180.2.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dolan S. A., Miller L. H., Wellems T. E. Evidence for a switching mechanism in the invasion of erythrocytes by Plasmodium falciparum. J Clin Invest. 1990 Aug;86(2):618–624. doi: 10.1172/JCI114753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dolan S. A., Proctor J. L., Alling D. W., Okubo Y., Wellems T. E., Miller L. H. Glycophorin B as an EBA-175 independent Plasmodium falciparum receptor of human erythrocytes. Mol Biochem Parasitol. 1994 Mar;64(1):55–63. doi: 10.1016/0166-6851(94)90134-1. [DOI] [PubMed] [Google Scholar]
  10. Fang X. D., Kaslow D. C., Adams J. H., Miller L. H. Cloning of the Plasmodium vivax Duffy receptor. Mol Biochem Parasitol. 1991 Jan;44(1):125–132. doi: 10.1016/0166-6851(91)90228-x. [DOI] [PubMed] [Google Scholar]
  11. Galinski M. R., Barnwell J. W. Plasmodium vivax: Merozoites, invasion of reticulocytes and considerations for malaria vaccine development. Parasitol Today. 1996 Jan;12(1):20–29. doi: 10.1016/0169-4758(96)80641-7. [DOI] [PubMed] [Google Scholar]
  12. Galinski M. R., Medina C. C., Ingravallo P., Barnwell J. W. A reticulocyte-binding protein complex of Plasmodium vivax merozoites. Cell. 1992 Jun 26;69(7):1213–1226. doi: 10.1016/0092-8674(92)90642-p. [DOI] [PubMed] [Google Scholar]
  13. Gibson H. L., Tucker J. E., Kaslow D. C., Krettli A. U., Collins W. E., Kiefer M. C., Bathurst I. C., Barr P. J. Structure and expression of the gene for Pv200, a major blood-stage surface antigen of Plasmodium vivax. Mol Biochem Parasitol. 1992 Feb;50(2):325–333. doi: 10.1016/0166-6851(92)90230-h. [DOI] [PubMed] [Google Scholar]
  14. Gupta S., Trenholme K., Anderson R. M., Day K. P. Antigenic diversity and the transmission dynamics of Plasmodium falciparum. Science. 1994 Feb 18;263(5149):961–963. doi: 10.1126/science.8310293. [DOI] [PubMed] [Google Scholar]
  15. Hadley T. J., Klotz F. W., Pasvol G., Haynes J. D., McGinniss M. H., Okubo Y., Miller L. H. Falciparum malaria parasites invade erythrocytes that lack glycophorin A and B (MkMk). Strain differences indicate receptor heterogeneity and two pathways for invasion. J Clin Invest. 1987 Oct;80(4):1190–1193. doi: 10.1172/JCI113178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jakobsen P. H., Hviid L., Theander T. G., Afare E. A., Ridley R. G., Heegaard P. M., Stuber D., Dalsgaard K., Nkrumah F. K. Specific T-cell recognition of the merozoite proteins rhoptry-associated protein 1 and erythrocyte-binding antigen 1 of Plasmodium falciparum. Infect Immun. 1993 Jan;61(1):268–273. doi: 10.1128/iai.61.1.268-273.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kaslow D. C., Kumar S. Expression and immunogenicity of the C-terminus of a major blood-stage surface protein of Plasmodium vivax, Pv200(19), secreted from Saccharomyces cerevisiae. Immunol Lett. 1996 Jul;51(3):187–189. doi: 10.1016/0165-2478(96)02570-9. [DOI] [PubMed] [Google Scholar]
  18. Miller L. H., Mason S. J., Clyde D. F., McGinniss M. H. The resistance factor to Plasmodium vivax in blacks. The Duffy-blood-group genotype, FyFy. N Engl J Med. 1976 Aug 5;295(6):302–304. doi: 10.1056/NEJM197608052950602. [DOI] [PubMed] [Google Scholar]
  19. Miller L. H., Powers K. G., Shiroishi T. Plasmodium knowlesi: functional immunity and antimerozoite antibodies in rhesus monkeys after repeated infection. Exp Parasitol. 1977 Feb;41(1):105–111. doi: 10.1016/0014-4894(77)90135-7. [DOI] [PubMed] [Google Scholar]
  20. Mitchell G. H., Hadley T. J., McGinniss M. H., Klotz F. W., Miller L. H. Invasion of erythrocytes by Plasmodium falciparum malaria parasites: evidence for receptor heterogeneity and two receptors. Blood. 1986 May;67(5):1519–1521. [PubMed] [Google Scholar]
  21. Perkins M. E., Holt E. H. Erythrocyte receptor recognition varies in Plasmodium falciparum isolates. Mol Biochem Parasitol. 1988 Jan 1;27(1):23–34. doi: 10.1016/0166-6851(88)90021-7. [DOI] [PubMed] [Google Scholar]
  22. Peterson D. S., Miller L. H., Wellems T. E. Isolation of multiple sequences from the Plasmodium falciparum genome that encode conserved domains homologous to those in erythrocyte-binding proteins. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):7100–7104. doi: 10.1073/pnas.92.15.7100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sim B. K., Chitnis C. E., Wasniowska K., Hadley T. J., Miller L. H. Receptor and ligand domains for invasion of erythrocytes by Plasmodium falciparum. Science. 1994 Jun 24;264(5167):1941–1944. doi: 10.1126/science.8009226. [DOI] [PubMed] [Google Scholar]
  24. Sim B. K., Orlandi P. A., Haynes J. D., Klotz F. W., Carter J. M., Camus D., Zegans M. E., Chulay J. D. Primary structure of the 175K Plasmodium falciparum erythrocyte binding antigen and identification of a peptide which elicits antibodies that inhibit malaria merozoite invasion. J Cell Biol. 1990 Nov;111(5 Pt 1):1877–1884. doi: 10.1083/jcb.111.5.1877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smith J. D., Chitnis C. E., Craig A. G., Roberts D. J., Hudson-Taylor D. E., Peterson D. S., Pinches R., Newbold C. I., Miller L. H. Switches in expression of Plasmodium falciparum var genes correlate with changes in antigenic and cytoadherent phenotypes of infected erythrocytes. Cell. 1995 Jul 14;82(1):101–110. doi: 10.1016/0092-8674(95)90056-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Su X. Z., Heatwole V. M., Wertheimer S. P., Guinet F., Herrfeldt J. A., Peterson D. S., Ravetch J. A., Wellems T. E. The large diverse gene family var encodes proteins involved in cytoadherence and antigenic variation of Plasmodium falciparum-infected erythrocytes. Cell. 1995 Jul 14;82(1):89–100. doi: 10.1016/0092-8674(95)90055-1. [DOI] [PubMed] [Google Scholar]
  27. Tsuboi T., Kappe S. H., al-Yaman F., Prickett M. D., Alpers M., Adams J. H. Natural variation within the principal adhesion domain of the Plasmodium vivax duffy binding protein. Infect Immun. 1994 Dec;62(12):5581–5586. doi: 10.1128/iai.62.12.5581-5586.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wertheimer S. P., Barnwell J. W. Plasmodium vivax interaction with the human Duffy blood group glycoprotein: identification of a parasite receptor-like protein. Exp Parasitol. 1989 Nov;69(4):340–350. doi: 10.1016/0014-4894(89)90083-0. [DOI] [PubMed] [Google Scholar]

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