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. 1990 Apr;58(4):854–859. doi: 10.1128/iai.58.4.854-859.1990

Immunogenicity and antigenicity in rabbits of a repeated sequence of Plasmodium falciparum antigen Pf155/RESA fused to two immunoglobulin G-binding domains of staphylococcal protein A.

A Sjölander 1, S Ståhl 1, P A Nygren 1, L Aslund 1, N Ahlborg 1, B Wåhlin 1, A Scherf 1, K Berzins 1, M Uhlén 1, P Perlmann 1
PMCID: PMC258551  PMID: 2180822

Abstract

A synthetic gene encoding a tetramer of the repeated subunit EENVEHDA of the Plasmodium falciparum antigen Pf155/RESA was expressed in a dual-expression system. The resulting fusion proteins, designated ZZ-M1 and BB-M1, comprised the EENVEHDA repeats and either two immunoglobulin G-binding domains from staphylococcal protein A or the human serum albumin-binding domains from streptococcal protein G, respectively. The soluble fusion proteins were affinity purified to homogeneity in one-step procedures. ZZ-M1 was used for immunization of rabbits. The rabbit antisera reacted with BB-M1 in an enzyme-linked immunosorbent assay and with Pf155/RESA in immunofluorescence of infected erythrocytes and immunoblotting. Inhibition studies revealed that the antibodies mainly recognized epitopes formed by two or more EENVEHDA subunits and were remarkably specific for Pf155/RESA. Importantly, the antibodies also inhibited P. falciparum merozoite reinvasion in vitro efficiently, indicating that they reacted with biologically important epitopes exposed on the native antigen. Immunization with Freund complete adjuvant resulted in high levels of specific immunoglobulin G antibodies over a 1-year period, whereas the antibody response obtained after immunization without adjuvant was generally weaker, immunoglobulin G and M mediated, and not sustained for longer periods. However, these titers were restored after booster injection. Taken together, the results support the usefulness of recombinant gene constructs of this type as immunogens for malaria vaccines.

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Selected References

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  1. Aslund L., Sjölander A., Wahlgren M., Wåhlin B., Ruangjirachuporn W., Berzins K., Wigzell H., Perlmann P., Pettersson U. Synthetic gene construct expressing a repeated and highly immunogenic epitope of the Plasmodium falciparum antigen Pf155. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1399–1403. doi: 10.1073/pnas.84.5.1399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berzins K., Perlmann H., Wåhlin B., Carlsson J., Wahlgren M., Udomsangpetch R., Björkman A., Patarroyo M. E., Perlmann P. Rabbit and human antibodies to a repeated amino acid sequence of a Plasmodium falciparum antigen, Pf 155, react with the native protein and inhibit merozoite invasion. Proc Natl Acad Sci U S A. 1986 Feb;83(4):1065–1069. doi: 10.1073/pnas.83.4.1065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berzins K., Wahlgren M., Perlmann P. Studies on the specificity of anti-erythrocyte antibodies in the serum of patients with malaria. Clin Exp Immunol. 1983 Nov;54(2):313–318. [PMC free article] [PubMed] [Google Scholar]
  4. Braesch-Andersen S., Paulie S., Koho H., Perlmann P. Isolation and characterization of two bladder carcinoma-associated antigens. J Immunol Methods. 1986 Nov 20;94(1-2):145–151. doi: 10.1016/0022-1759(86)90227-9. [DOI] [PubMed] [Google Scholar]
  5. Collins W. E., Anders R. F., Pappaioanou M., Campbell G. H., Brown G. V., Kemp D. J., Coppel R. L., Skinner J. C., Andrysiak P. M., Favaloro J. M. Immunization of Aotus monkeys with recombinant proteins of an erythrocyte surface antigen of Plasmodium falciparum. Nature. 1986 Sep 18;323(6085):259–262. doi: 10.1038/323259a0. [DOI] [PubMed] [Google Scholar]
  6. Coppel R. L., Cowman A. F., Anders R. F., Bianco A. E., Saint R. B., Lingelbach K. R., Kemp D. J., Brown G. V. Immune sera recognize on erythrocytes Plasmodium falciparum antigen composed of repeated amino acid sequences. 1984 Aug 30-Sep 5Nature. 310(5980):789–792. doi: 10.1038/310789a0. [DOI] [PubMed] [Google Scholar]
  7. Favaloro J. M., Coppel R. L., Corcoran L. M., Foote S. J., Brown G. V., Anders R. F., Kemp D. J. Structure of the RESA gene of Plasmodium falciparum. Nucleic Acids Res. 1986 Nov 11;14(21):8265–8277. doi: 10.1093/nar/14.21.8265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kabilan L., Troye-Blomberg M., Perlmann H., Andersson G., Högh B., Petersen E., Björkman A., Perlmann P. T-cell epitopes in Pf155/RESA, a major candidate for a Plasmodium falciparum malaria vaccine. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5659–5663. doi: 10.1073/pnas.85.15.5659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Löwenadler B., Jansson B., Paleus S., Holmgren E., Nilsson B., Moks T., Palm G., Josephson S., Philipson L., Uhlén M. A gene fusion system for generating antibodies against short peptides. Gene. 1987;58(1):87–97. doi: 10.1016/0378-1119(87)90032-1. [DOI] [PubMed] [Google Scholar]
  10. Löwenadler B., Nilsson B., Abrahmsén L., Moks T., Ljungqvist L., Holmgren E., Paleus S., Josephson S., Philipson L., Uhlén M. Production of specific antibodies against protein A fusion proteins. EMBO J. 1986 Sep;5(9):2393–2398. doi: 10.1002/j.1460-2075.1986.tb04509.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Miller L. H., Howard R. J., Carter R., Good M. F., Nussenzweig V., Nussenzweig R. S. Research toward malaria vaccines. Science. 1986 Dec 12;234(4782):1349–1356. doi: 10.1126/science.2431481. [DOI] [PubMed] [Google Scholar]
  12. Nossal N. G., Heppel L. A. The release of enzymes by osmotic shock from Escherichia coli in exponential phase. J Biol Chem. 1966 Jul 10;241(13):3055–3062. [PubMed] [Google Scholar]
  13. Nygren P. A., Eliasson M., Abrahmsén L., Uhlén M., Palmcrantz E. Analysis and use of the serum albumin binding domains of streptococcal protein G. J Mol Recognit. 1988 Apr;1(2):69–74. doi: 10.1002/jmr.300010204. [DOI] [PubMed] [Google Scholar]
  14. Perlmann H., Berzins K., Wahlgren M., Carlsson J., Björkman A., Patarroyo M. E., Perlmann P. Antibodies in malarial sera to parasite antigens in the membrane of erythrocytes infected with early asexual stages of Plasmodium falciparum. J Exp Med. 1984 Jun 1;159(6):1686–1704. doi: 10.1084/jem.159.6.1686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Perlmann H., Perlmann P., Berzins K., Wåhlin B., Troye-Blomberg M., Hagstedt M., Andersson I., Högh B., Petersen E., Björkman A. Dissection of the human antibody response to the malaria antigen Pf155/RESA into epitope specific components. Immunol Rev. 1989 Dec;112:115–132. doi: 10.1111/j.1600-065x.1989.tb00555.x. [DOI] [PubMed] [Google Scholar]
  16. Perlmann P., Berzins K., Perlmann H., Troye-Blomberg M., Wahlgren M., Wåhlin B. Malaria vaccines: immunogen selection and epitope mapping. Vaccine. 1988 Apr;6(2):183–187. doi: 10.1016/s0264-410x(88)80025-2. [DOI] [PubMed] [Google Scholar]
  17. Rzepczyk C. M., Ramasamy R., Ho P. C., Mutch D. A., Anderson K. L., Duggleby R. G., Doran T. J., Murray B. J., Irving D. O., Woodrow G. C. Identification of T epitopes within a potential Plasmodium falciparum vaccine antigen. A study of human lymphocyte responses to repeat and nonrepeat regions of Pf155/RESA. J Immunol. 1988 Nov 1;141(9):3197–3202. [PubMed] [Google Scholar]
  18. Scherf A., Hilbich C., Sieg K., Mattei D., Mercereau-Puijalon O., Müller-Hill B. The 11-1 gene of Plasmodium falciparum codes for distinct fast evolving repeats. EMBO J. 1988 Apr;7(4):1129–1137. doi: 10.1002/j.1460-2075.1988.tb02922.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ståhl S., Sjölander A., Nygren P. A., Berzins K., Perlmann P., Uhlén M. A dual expression system for the generation, analysis and purification of antibodies to a repeated sequence of the Plasmodium falciparum antigen Pf155/RESA. J Immunol Methods. 1989 Nov 13;124(1):43–52. doi: 10.1016/0022-1759(89)90184-1. [DOI] [PubMed] [Google Scholar]
  20. Udomsangpetch R., Carlsson J., Wåhlin B., Holmquist G., Ozaki L. S., Scherf A., Mattei D., Mercereau-Puijalon O., Uni S., Aikawa M. Reactivity of the human monoclonal antibody 33G2 with repeated sequences of three distinct Plasmodium falciparum antigens. J Immunol. 1989 May 15;142(10):3620–3626. [PubMed] [Google Scholar]
  21. Wahlgren M., Berzins K., Perlmann P., Björkman A. Characterization of the humoral immune response in Plasmodium falciparum malaria. I. Estimation of antibodies to P. falciparum or human erythrocytes by means of microELISA. Clin Exp Immunol. 1983 Oct;54(1):127–134. [PMC free article] [PubMed] [Google Scholar]
  22. Wåhlin B., Wahlgren M., Perlmann H., Berzins K., Björkman A., Patarroyo M. E., Perlmann P. Human antibodies to a Mr 155,000 Plasmodium falciparum antigen efficiently inhibit merozoite invasion. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7912–7916. doi: 10.1073/pnas.81.24.7912. [DOI] [PMC free article] [PubMed] [Google Scholar]

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