Li Wenshu, Chen Qingxin, Lin Qiaoai, Lv Yan, Feng Juan, Liu Jianxiao, Xu Wen, Chen Shao, Zhu Xiaochun, Zhang Lifang
Institute of Molecular Virology and Immunology, Department of Microbiology and Immunology, Wenzhou Medical College, Wenzhou, 325035, Zhejiang Province, China.
Protein Pept Lett. 2013 Oct;20(10):1136-43. doi: 10.2174/09298665113209990005.
Epstein-Barr virus (EBV) is a human oncogenic herpesvirus associating with several malignant diseases. Latent membrane protein 2 (LMP2) of EBV is considered to be an ideal candidate for immunotherapy or prophylactic EBV vaccine. We designed a LMP2 multiepitope containing T and B-cell epitope-rich peptides and constructed a recombinant plasmid containing mammalian codonoptimization EBV LMP2 multiepitope (pcDNA3.1+/EBV-LMP2 multiepitope). After pcDNA3.1+/EBV-LMP2 multiepitope was transfected into COS-7 cells, significant expression of the multiepitope in COS-7 cells was confirmed by RT-PCR and immunofluorescence assay. Western blot analysis indicated that serum from immunized mice could be discerned by the EBV-LMP2 protein and the EBV-LMP2 multiepitope specifically. The plasmid DNA of EBV-LMP2 multiepitope induced high levels anti-EBV membrane protein and anti-EBV LMP2 multiepitope IgG in mice. T lymphocytes from spleen of immunized mice showed a strong CTL activity. The present study suggested that plasmid DNA encoding EBV LMP2 multiepitope capable of stimulating enough cellular and humoral immunity could have potential for preventing or controlling EBV infection and EBV associated disease in mice.
爱泼斯坦-巴尔病毒（EBV）是一种与多种恶性疾病相关的人类致癌性疱疹病毒。EBV的潜伏膜蛋白2（LMP2）被认为是免疫治疗或预防性EBV疫苗的理想候选物。我们设计了一种包含富含T细胞和B细胞表位的肽的LMP2多表位，并构建了一个含有经哺乳动物密码子优化的EBV LMP2多表位的重组质粒（pcDNA3.1+/EBV-LMP2多表位）。将pcDNA3.1+/EBV-LMP2多表位转染到COS-7细胞后，通过RT-PCR和免疫荧光测定证实了该多表位在COS-7细胞中的显著表达。蛋白质印迹分析表明，免疫小鼠的血清可以被EBV-LMP2蛋白和EBV-LMP2多表位特异性识别。EBV-LMP2多表位的质粒DNA在小鼠中诱导了高水平的抗EBV膜蛋白和抗EBV LMP2多表位IgG。免疫小鼠脾脏中的T淋巴细胞表现出很强的CTL活性。本研究表明，编码EBV LMP2多表位的质粒DNA能够刺激足够的细胞免疫和体液免疫，在预防或控制小鼠的EBV感染和EBV相关疾病方面可能具有潜力。
