Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1993 Mar;61(3):892–897. doi: 10.1128/iai.61.3.892-897.1993

Conservation of antigen components from two recombinant hybrid proteins protective against malaria.

B Knapp 1, U Nau 1, E Hundt 1
PMCID: PMC302816  PMID: 8432609

Abstract

Recently, we have shown that two hybrid proteins carrying partial sequences of the blood-stage antigens SERP, HRPII, and MSAI from Plasmodium falciparum confer protective immunity on Aotus monkeys against an experimental parasite infection (B. Knapp, E. Hundt, B. Enders, and H. A. Küpper, Infect. Immun. 60:2397-2401, 1992). The malarial components of the hybrid proteins consist of amino acid residues 630 to 892 of SERP, amino acid residues 146 to 260 of MSAI, and the 189 C-terminal residues of HRPII. We have studied the diversity of these protein regions in field isolates of P. falciparum. Genomic DNA was extracted from the blood of six donors from two different areas where malaria is endemic. The gene regions of SERP and MSAI coding for the corresponding sequences of the protective hybrid proteins and the exon II region of the HRPII gene were amplified by polymerase chain reaction and sequenced. All three regions were found to be highly conserved. In the 262-amino-acid fragment of SERP, one single conservative amino acid substitution was found. The exon II region of HRPII showed only a slight variability in number and arrangement of the repeat units. The 115-amino-acid fragment of MSAI which is located within an N-terminal region known to be conserved among different parasite strains was shown to be the most variable among the vaccine components: amino acid substitutions were found in 14 different positions of this MSAI region when both laboratory strains and field isolates were compared.

Full text

PDF
892

Selected References

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

  1. Bzik D. J., Li W. B., Horii T., Inselburg J. Amino acid sequence of the serine-repeat antigen (SERA) of Plasmodium falciparum determined from cloned cDNA. Mol Biochem Parasitol. 1988 Sep;30(3):279–288. doi: 10.1016/0166-6851(88)90097-7. [DOI] [PubMed] [Google Scholar]
  2. Casanova J. L., Pannetier C., Jaulin C., Kourilsky P. Optimal conditions for directly sequencing double-stranded PCR products with sequenase. Nucleic Acids Res. 1990 Jul 11;18(13):4028–4028. doi: 10.1093/nar/18.13.4028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Certa U., Rotmann D., Matile H., Reber-Liske R. A naturally occurring gene encoding the major surface antigen precursor p190 of Plasmodium falciparum lacks tripeptide repeats. EMBO J. 1987 Dec 20;6(13):4137–4142. doi: 10.1002/j.1460-2075.1987.tb02759.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chang S. P., Kramer K. J., Yamaga K. M., Kato A., Case S. E., Siddiqui W. A. Plasmodium falciparum: gene structure and hydropathy profile of the major merozoite surface antigen (gp195) of the Uganda-Palo Alto isolate. Exp Parasitol. 1988 Oct;67(1):1–11. doi: 10.1016/0014-4894(88)90002-1. [DOI] [PubMed] [Google Scholar]
  5. Cheung A., Leban J., Shaw A. R., Merkli B., Stocker J., Chizzolini C., Sander C., Perrin L. H. Immunization with synthetic peptides of a Plasmodium falciparum surface antigen induces antimerozoite antibodies. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8328–8332. doi: 10.1073/pnas.83.21.8328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cheung A., Shaw A. R., Leban J., Perrin L. H. Cloning and expression in Escherichia coli of a surface antigen of Plasmodium falciparum merozoites. EMBO J. 1985 Apr;4(4):1007–1011. doi: 10.1002/j.1460-2075.1985.tb03731.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coppel R. L., Crewther P. E., Culvenor J. G., Perrin L. H., Brown G. V., Kemp D. J., Anders R. F. Variation in p126, a parasitophorous vacuole antigen of Plasmodium falciparum. Mol Biol Med. 1988 Dec;5(3):155–166. [PubMed] [Google Scholar]
  8. Crisanti A., Müller H. M., Hilbich C., Sinigaglia F., Matile H., McKay M., Scaife J., Beyreuther K., Bujard H. Epitopes recognized by human T cells map within the conserved part of the GP190 of P. falciparum. Science. 1988 Jun 3;240(4857):1324–1326. doi: 10.1126/science.2453924. [DOI] [PubMed] [Google Scholar]
  9. Cysteine or serine proteinase? Nature. 1989 Nov 9;342(6246):132–132. doi: 10.1038/342132b0. [DOI] [PubMed] [Google Scholar]
  10. Delplace P., Bhatia A., Cagnard M., Camus D., Colombet G., Debrabant A., Dubremetz J. F., Dubreuil N., Prensier G., Fortier B. Protein p126: a parasitophorous vacuole antigen associated with the release of Plasmodium falciparum merozoites. Biol Cell. 1988;64(2):215–221. doi: 10.1016/0248-4900(88)90080-9. [DOI] [PubMed] [Google Scholar]
  11. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guttinger M., Romagnoli P., Vandel L., Meloen R., Takacs B., Pink J. R., Sinigaglia F. HLA polymorphism and T cell recognition of a conserved region of p190, a malaria vaccine candidate. Int Immunol. 1991 Sep;3(9):899–906. doi: 10.1093/intimm/3.9.899. [DOI] [PubMed] [Google Scholar]
  13. Herrera M. A., Rosero F., Herrera S., Caspers P., Rotmann D., Sinigaglia F., Certa U. Protection against malaria in Aotus monkeys immunized with a recombinant blood-stage antigen fused to a universal T-cell epitope: correlation of serum gamma interferon levels with protection. Infect Immun. 1992 Jan;60(1):154–158. doi: 10.1128/iai.60.1.154-158.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Higgins D. G., McConnell D. J., Sharp P. M. Malarial proteinase? Nature. 1989 Aug 24;340(6235):604–604. doi: 10.1038/340604a0. [DOI] [PubMed] [Google Scholar]
  15. Holder A. A., Lockyer M. J., Odink K. G., Sandhu J. S., Riveros-Moreno V., Nicholls S. C., Hillman Y., Davey L. S., Tizard M. L., Schwarz R. T. Primary structure of the precursor to the three major surface antigens of Plasmodium falciparum merozoites. Nature. 1985 Sep 19;317(6034):270–273. doi: 10.1038/317270a0. [DOI] [PubMed] [Google Scholar]
  16. Inselburg J., Bzik D. J., Li W. B., Green K. M., Kansopon J., Hahm B. K., Bathurst I. C., Barr P. J., Rossan R. N. Protective immunity induced in Aotus monkeys by recombinant SERA proteins of Plasmodium falciparum. Infect Immun. 1991 Apr;59(4):1247–1250. doi: 10.1128/iai.59.4.1247-1250.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jongwutiwes S., Tanabe K., Nakazawa S., Yanagi T., Kanbara H. Sequence variation in the tripeptide repeats and T cell epitopes in P190 (MSA-1) of Plasmodium falciparum from field isolates. Mol Biochem Parasitol. 1992 Mar;51(1):81–89. doi: 10.1016/0166-6851(92)90203-v. [DOI] [PubMed] [Google Scholar]
  18. Kimura E., Mattei D., di Santi S. M., Scherf A. Genetic diversity in the major merozoite surface antigen of Plasmodium falciparum: high prevalence of a third polymorphic form detected in strains derived from malaria patients. Gene. 1990 Jul 2;91(1):57–62. doi: 10.1016/0378-1119(90)90162-k. [DOI] [PubMed] [Google Scholar]
  19. Knapp B., Hundt E., Enders B., Küpper H. A. A recombinant hybrid protein as antigen for an anti-blood stage malaria vaccine. Behring Inst Mitt. 1991 Feb;(88):147–156. [PubMed] [Google Scholar]
  20. 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]
  21. Knapp B., Hundt E., Nau U., Küpper H. A. Molecular cloning, genomic structure and localization in a blood stage antigen of Plasmodium falciparum characterized by a serine stretch. Mol Biochem Parasitol. 1989 Jan 1;32(1):73–83. doi: 10.1016/0166-6851(89)90131-x. [DOI] [PubMed] [Google Scholar]
  22. Knapp B., Shaw A., Hundt E., Enders B., Küpper H. A. A histidin alanine rich recombinant antigen protects Aotus monkeys from P. falciparum infection. Behring Inst Mitt. 1988 Apr;(82):349–359. [PubMed] [Google Scholar]
  23. Mackay M., Goman M., Bone N., Hyde J. E., Scaife J., Certa U., Stunnenberg H., Bujard H. Polymorphism of the precursor for the major surface antigens of Plasmodium falciparum merozoites: studies at the genetic level. EMBO J. 1985 Dec 30;4(13B):3823–3829. doi: 10.1002/j.1460-2075.1985.tb04154.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Perrin L. H., Merkli B., Loche M., Chizzolini C., Smart J., Richle R. Antimalarial immunity in Saimiri monkeys. Immunization with surface components of asexual blood stages. J Exp Med. 1984 Aug 1;160(2):441–451. doi: 10.1084/jem.160.2.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Peterson M. G., Coppel R. L., McIntyre P., Langford C. J., Woodrow G., Brown G. V., Anders R. F., Kemp D. J. Variation in the precursor to the major merozoite surface antigens of Plasmodium falciparum. Mol Biochem Parasitol. 1988 Jan 15;27(2-3):291–301. doi: 10.1016/0166-6851(88)90049-7. [DOI] [PubMed] [Google Scholar]
  26. Roussilhon C., Hundt E., Agrapart M., Stüber W., Knapp B., Dubois P., Ballet J. J. Responses of T cells from sensitized donors to recombinant and synthetic peptides corresponding to sequences of the Plasmodium falciparum SERP antigen. Immunol Lett. 1990 Aug;25(1-3):149–153. doi: 10.1016/0165-2478(90)90107-2. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Snewin V. A., Herrera M., Sanchez G., Scherf A., Langsley G., Herrera S. Polymorphism of the alleles of the merozoite surface antigens MSA1 and MSA2 in Plasmodium falciparum wild isolates from Colombia. Mol Biochem Parasitol. 1991 Dec;49(2):265–275. doi: 10.1016/0166-6851(91)90070-m. [DOI] [PubMed] [Google Scholar]
  29. Tanabe K., Mackay M., Goman M., Scaife J. G. Allelic dimorphism in a surface antigen gene of the malaria parasite Plasmodium falciparum. J Mol Biol. 1987 May 20;195(2):273–287. doi: 10.1016/0022-2836(87)90649-8. [DOI] [PubMed] [Google Scholar]
  30. Weber J. L., Leininger W. M., Lyon J. A. Variation in the gene encoding a major merozoite surface antigen of the human malaria parasite Plasmodium falciparum. Nucleic Acids Res. 1986 Apr 25;14(8):3311–3323. doi: 10.1093/nar/14.8.3311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wellems T. E., Howard R. J. Homologous genes encode two distinct histidine-rich proteins in a cloned isolate of Plasmodium falciparum. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6065–6069. doi: 10.1073/pnas.83.16.6065. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES