Construction and immunogenicity prediction of Plasmodiumfalciparum CTL epitope minigene vaccine
- 20 Downloads
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.
KeywordsPlasmodiumfalciparum CD8+ cytotoxic T cell MHC class I molecule
Unable to display preview. Download preview PDF.
- 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.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
- 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