Construction of specifically targeted chimeric hepatitis E virus DNA vaccine and its efficacy assessment.
OBJECTIVE: To design chimeric DNA vaccine targeted to antigen-presenting cells (APCs) with enhanced efficacy to induce immunization. METHODS: The plasmid containing the gene encoding cytotoxic T-lymphocyte antigen 4 (CTLA4), a surface molecule on T cells, was directly fused to the gene fragment HEVE2 coding for hepatitis E virus antigen by molecular engineering technology. The plasmid containing HEVE2 gene fragment alone was also constructed to serve as control and transfection of COS-7 cells with the 2 resultant plasmids was performed respectively, followed by assay of the expressions of CTLA4- HEVE2 fusion protein and HEVE2 protein in COS-7 cells by way of Western blotting. BALB/c mice were injected intracutaneously with the DNA vaccine (100 microgram) for 3 times at 2-week intervals, and the titer of anti-HEVE2 total IgG and IgG subclasses were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The mammalian expression plasmids of CTLA4-HEVE2 fusion protein or HEVE2 protein alone were cloned. The culture supernants of COS-7 cells transfected with the plasmids showed the production of CTLA4-HEVE2 and HEVE2 proteins, which were secreted in the form of dimers. Mice immunized with pCTLA2-HEVE2 produced high levels of specific anti-HEVE2 total IgG titers with IgG2a, IgG2b and IgG1 subclasses predominant in the serum, approximately 50- to 100-fold higher than those in mice immunized with pHEVE2, whose serum contained predominantly IgG1. CONCLUSION: CTLA4-HEVE2 chimeric vaccine stimulates strong immune responses in mice, making it possible for further exploration into chimeric DNA vaccines that target the antigen to APCs.[1]References
- Construction of specifically targeted chimeric hepatitis E virus DNA vaccine and its efficacy assessment. Che, X.Y., Qian, Q.J., Liu, S.X., Huang, J., Qiu, Q.L., Xu, H., Wu, W.H. Di Yi Jun Yi Da Xue Xue Bao (2002) [Pubmed]
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