期刊
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
卷 97, 期 14, 页码 6251-6262出版社
SPRINGER
DOI: 10.1007/s00253-013-4967-x
关键词
Epstein-Barr virus (EBV); EBNA1; Pichia pastoris; Immunogenicity; Polyclonal antibodies; Vaccine
资金
- National Basic Research Program of China (973 Program) [2011CB504800]
Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus associated with the development of both lymphoid and epithelial tumors. EBNA1 is the only viral protein expressed in all EBV-associated malignancies and plays important roles in EBV latency. Thus, EBNA1 is thought to be a promising antigen for immunotherapy of all EBV-associated malignancies. This study was undertaken to produce recombinant EBNA1 protein in Pichia pastoris and evaluate its immunogenicity. The truncated EBNA1 (E1 Delta GA, codons 390-641) was expressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast P. pastoris and purified by Ni-NTA affinity chromatography. The purified proteins were then used as antigens to immunize BALB/c mice for production of polyclonal antibodies. Western blot analysis showed that the polyclonal antibodies specifically recognized the EBNA1 protein in B95-8 cell lysates. The recombinant E1 Delta GA also induced strong lymphoproliferative and Th1 cytokine responses in mice. Furthermore, mice immunized with E1 Delta GA developed CD4(+) and CD8(+) T cell responses. These findings showed that the yeast-expressed E1 Delta GA retained good immunogenicity and might be a promising vaccine candidate against EBV-associated malignancies.
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