Science in China Series C: Life Sciences

, Volume 44, Issue 2, pp 207–215 | Cite as

Construction and immunogenicity prediction of Plasmodiumfalciparum CTL epitope minigene vaccine



The minigenes encoding Plasmodiumfalciparum CTL epitopes restricted to human MHC class I molecular HLA-A2 and HLA-B51, which were both at high frequency among Chinese population, were constructed as mono-epitope CTL vaccines named pcDNA3.1/tr and pcDNA3.1/sh. The minigenes of the two epitopes were then tandem linked to form a dimeric CTL epitope minigene recombinant vaccine. After DNA transfection, the epitope minigenes were expressed respectively in two human cell lines, each bearing one MHC class I molecule named CIR/HLA-A2.1 and K562/HLA-B51. The intracellular expression of the CTL epitope minigenes not only enhanced the stability of HLA-A2.1 and HLA-B51 molecules but also increased the assemblage of MHC class I molecules on cell surfaces, which testified the specific process and presentation of those endogenous expressed epitopes. For the cells transfected with the dimeric minigene encoding two tandem linked epitopes, the expression and presentation of each epitope were also detected on cell membranes that bore different MHC class I molecules. It meant that the adjacency of the two CTL epitopes did not interfere with the specific process and presentation of each epitope. Compared with the ordinary CTL studies that inoculated synthesized epitope peptides with peripheral blood cells, this work aimed to process the epitopes directly inside HLA class I allele specific human cells, and thus theoretically imitated the same procedurein vivo. It was also an economical way to predict the immunogenicity of CTL epitopes at an early stage especially in laboratories with limited financial resource.


Plasmodiumfalciparum CD8+ cytotoxic T cell MHC class I molecule 


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  1. 1.
    Hill, A. V., Elvin, J., Willis, A. C. et al., Molecular analysis of the association of HLA-B53 and resistance to severe malaria, Nature, 1992, 360: 434.PubMedCrossRefGoogle Scholar
  2. 2.
    Perlmann, P., Miller, L., Fogarty/WHO international conference on cellular mechanisms in malaria immunity, Immun. Letter, 1990, 25: 1.CrossRefGoogle Scholar
  3. 3.
    Perkus, M. E., Tartaglia, J., Paoletti, E. et al., Poxvirus-based vaccine candidates for cancer, AIDS and other infectious diseases, J. Leukocyte Biol., 1995, 58(1): 1.PubMedGoogle Scholar
  4. 4.
    Shen, H., Slifka, M. K., Matloubian, M. et al., Recombinant Listeria monocytogenes as a live vaccine vehicle for the induction of protective anti-viral cell-mediated immunity, Proc. Natl. Acad. Sci. USA, 1995, 92: 3987.PubMedCrossRefGoogle Scholar
  5. 5.
    Waine, G. J., McManus, D. P., Nucleic acids: vaccines of the future, Parasitai Today, 1995, 11: 113.CrossRefGoogle Scholar
  6. 6.
    Whitton, J. L., Sheng, N., Oldstone, M. B. A. et al., A “string-of beads” vaccine, comprising linked minigenes, confers protection from lethal-dose virus challenge, J. Virol., 1993, 67: 348.PubMedGoogle Scholar
  7. 7.
    Lalvani, A., Aidoo, M., Allsopp, C. E. et al., An-HLA-based approach to design of a CTL-inducing vaccine against Plasmodiumfalciparum, Res. Immunol., 1994, 145: 461.PubMedCrossRefGoogle Scholar
  8. 8.
    Sidney, J., Grey, H. M., Kubo, R. T. et al., Practical, biochemical and evolutionary implications of the discovery of HLA class I supermotifs, Immunology Today, 1996, 17: 261.PubMedCrossRefGoogle Scholar
  9. 9.
    Thomson, S. A., Elliott, S. L., Sherritt, M. A. et al., Recombinant polyepitope vaccines for the delivery of multiple CD8 cytotoxic T cell epitopes, J. Immun., 1996, 157: 822.PubMedGoogle Scholar
  10. 10.
    Hanke, T., Schneider, J., Gilbert, S. C. et al., DNA multi-CTL epitope vaccines for HIV and Plasmodiumfalciparum: immunogenicity in mice, Vaccine, 1998, 16: 426.PubMedCrossRefGoogle Scholar
  11. 11.
    Townsend, A. R. M., Rothbard, J., Gotch, F. M. et al., The epitopes of influenza nucleoprotein recognized by cytotoxic T lymphocytes can be defined with short synthetic peptides, Cell, 1986, 44: 959.PubMedCrossRefGoogle Scholar
  12. 12.
    Ojcius, D. M., Abastado, J. P., Casrouge, A. et al., Dissociation of the peptide-MHC class I complex limits the binding rate of exogenous peptide, J. Immunol., 1993, 151: 6020.PubMedGoogle Scholar
  13. 13.
    Lin, C. T., Jiang, Y. F., Yin, B. et al., The application of isocaudamers cloning technique in construction of Plasmodiumfalciparum multi-valent vaccine, J. Biochem. & Biophys. (in Chinese), 1999, 15: 973.Google Scholar
  14. 14.
    Goligan, L. I., Kruisbeek, A. M., Margulies, D. H. et al., Current Protocols in Immunology, New York: Greene Publishing Associates and Wiley-Interscience, 1991, 9.0.1–9.4.11.Google Scholar
  15. 15.
    Zemmour, J., Little, A. M., Schendel, D. et al., The HLA-A, B, C “negative” mutant cell line C1R expresses a novel HLA-B35 allele, which also has a point mutations in the translation initiation codon, J. Immunol., 1992, 148: 1941.PubMedGoogle Scholar
  16. 16.
    Van der Burg, S. H., Visseren, M. J. W., Brandt, R. M. P. et al., Immunogenicity of peptides bound to MHC class I molecules depends on the MHC-peptide complex stability, J. Immunol., 1996, 156: 3308.PubMedGoogle Scholar
  17. 17.
    Barnstable, C. J., Bodmer, W. F., Brown, G. et al., Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens—new tools for genetic analysis, Cell, 1978, 14(1): 9.PubMedCrossRefGoogle Scholar
  18. 18.
    Aidoo, M., Lalvani, A., Allsopp, C. E. M. et al., Identification of conserved antigenic components for a cytotoxic T lymphocyte-inducing vaccine against malaria, Lancet, 1995, 345: 1003.PubMedCrossRefGoogle Scholar
  19. 19.
    Feltkamp, M. C. W., Vierboom, M. P. M., Kast, W. M. et al., Efficient MHC class I-peptide binding is required but does not ensure MHC class I-restricted immunogenicity, Mol. Immunol., 1994, 31: 1391.PubMedCrossRefGoogle Scholar
  20. 20.
    Sette, A., Vitiello, A., Reherman, B. et al., The relationship between class I binding affinity and immunogenicity of potential cytotoxic T cell epitopes, J. Immunol., 1994, 153: 5586.PubMedGoogle Scholar
  21. 21.
    Del, V. M. D., Schlicht, H. J., Ruppert, T. et al., Efficient processing of an antigenic sequence for presentation by MHC class I molecules depends on its neighboring residues in the protein, Cell, 1990, 66: 1145.CrossRefGoogle Scholar
  22. 22.
    Cornelia, C., Bergmann, Yao, Q. et al., Flanking residues alter antigenicity and immunogenicity of multi-unit CTL epitopes, J. Immunol., 1996, 157: 3342.Google Scholar

Copyright information

© Science in China Press 2001

Authors and Affiliations

  1. 1.Institute of Basic Medical SciencesChinese Academy of Medical Sciences/School of Basic Medicine, Peking Union Medical CollegeBeijingChina

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