Single B or T-cell epitope-based DNA vaccine using modified vector induces specific immune response against hepadnavirus.
Epitope-based DNA vaccine is an effective and powerful approach against a variety of pathogens or tumors. In present study, we reconstructed a vector that could effectively express short B and T-cell epitope of duck/hepatitis B virus, and investigated the role of the epitope-based DNA vaccination. The pUC19 was modified by inserting the compact transient framework (CTF), including HCMV IE1 promoter, enhancer, Kozak sequence, dual stop codon and 3' terminal bovine growth hormone terminal signal and so on. This modified vector was designated pEC(K) and supposed to effectively express short peptide. A well-defined single B-cell and T-cell epitope encoding gene of duck/hepatitis B virus has been synthesized as candidate epitope and cloned into pEC(K) plasmid, respectively. Transfection of the recombinant DNA into C(2)C(12) cell showed that modified plasmid could effectively express both the single B-cell and T-cell short epitope in the culture supernatant as confirmed by dot immunoblot assay (DIA). The recombinant single B and T-cell epitope-based DNA vaccine was administrated to C57BL/6 mice and could greatly induce specific humoral and CTL response. In addition, the specific antibody against B epitope could specifically bind to the DHBV particles. This report demonstrated that single epitope-based DNA vaccine using modified plasmid vector pEC(K) could induce effective specific immune responses and could be of great use for DNA vaccines.
Xu
Huan-Bin
HB
Department of Immunology and Key Laboratory of Molecular Medicine of the Ministry of Education, Shanghai Medical college of Fudan University, P.R. China.
Xu
Wei
W
Chu
Yi-Wei
YW
Wang
Ying
Y
Xiong
Sidong
S
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004372:Ducks; D018985:Epitopes, B-Lymphocyte / Q000096:biosynthesis / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000096:biosynthesis / Q000276:immunology; D018347:Hepadnaviridae Infections / Q000276:immunology / Q000517:prevention & control; D006510:Hepatitis B Antibodies / Q000096:biosynthesis; D006512:Hepatitis B Core Antigens / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D015611:Hepatitis B Virus, Duck / Q000235:genetics / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids; D014162:Transfection; D014611:Vaccination; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000276:immunology*
D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D006510:Hepatitis B Antibodies; D006512:Hepatitis B Core Antigens; D006514:Hepatitis B Surface Antigens; D019444:Vaccines, DNA